bims-aporos Biomed News
on Apoptosis and reactive oxygen species
Issue of 2019‒08‒04
104 papers selected by
Gavin McStay
Staffordshire University


  1. Nanomaterials (Basel). 2019 Jul 29. pii: E1089. [Epub ahead of print]9(8):
    Gurunathan S, Jeyaraj M, Kang MH, Kim JH.
      Osteosarcoma (OS) is the most common type of cancer and the most frequent malignant bone tumor in childhood and adolescence. Nanomedicine has become an indispensable field in biomedical and clinical research, with nanoparticles (NPs) promising to increase the therapeutic efficacy of anticancer drugs. Doxorubicin (DOX) is a commonly used chemotherapeutic drug against OS; however, it causes severe side effects that restrict its clinical applications. Here, we investigated whether combining platinum NPs (PtNPs) and DOX could increase their anticancer activity in human bone OS epithelial cells (U2OS). PtNPs with nontoxic, effective, thermally stable, and thermoplasmonic properties were synthesized and characterized using tangeretin. We examined the combined effects of PtNPs and DOX on cell viability, proliferation, and morphology, reactive oxygen species (ROS) generation, lipid peroxidation, nitric oxide, protein carbonyl content, antioxidants, mitochondrial membrane potential (MMP), adenosine tri phosphate (ATP) level, apoptotic and antiapoptotic gene expression, oxidative stress-induced DNA damage, and DNA repair genes. PtNPs and DOX significantly inhibited U2OS viability and proliferation in a dose-dependent manner, increasing lactate dehydrogenase leakage, ROS generation, and malondialdehyde, nitric oxide, and carbonylated protein levels. Mitochondrial dysfunction was confirmed by reduced MMP, decreased ATP levels, and upregulated apoptotic/downregulated antiapoptotic gene expression. Oxidative stress was a major cause of cytotoxicity and genotoxicity, confirmed by decreased levels of various antioxidants. Furthermore, PtNPs and DOX increased 8-oxo-dG and 8-oxo-G levels and induced DNA damage and repair gene expression. Combination of cisplatin and DOX potentially induce apoptosis comparable to PtNPs and DOX. To the best of our knowledge, this is the first report to describe the combined effects of PtNPs and DOX in OS.
    Keywords:  DNA damage; apoptosis; cytotoxicity; doxorubicin; genotoxicity; oxidative stress; platinum nanoparticles
    DOI:  https://doi.org/10.3390/nano9081089
  2. Int J Biol Sci. 2019 ;15(8): 1637-1653
    Di S, Fan C, Ma Z, Li M, Guo K, Han D, Li X, Mu D, Yan X.
      Butein, a member of the chalcone family, is a potent anticarcinogen against multiple cancers, but its specific anti-NSCLC mechanism remains unknown. The present study examined the effects of butein treatment on NSCLC cell lines and NSCLC xenografts. Butein markedly decreased NSCLC cell viability; inhibited cell adhesion, migration, invasion, and colony forming ability; and induced cell apoptosis and G2/M phase arrest in NSCLC cells. Moreover, butein significantly inhibited PC-9 xenograft growth. Both in vivo and in vitro studies verified that butein exerted anti-NSCLC effect through activating endoplasmic reticulum (ER) stress-dependent reactive oxygen species (ROS) generation. These pro-apoptotic effects were reversed by the use of 4- phenylbutyric acid (4-PBA), CHOP siRNA, N-acetyl-L-cysteine (NAC) and Z-VAD-FMK (z-VAD) in vitro. Moreover, inhibition of ER stress markedly reduced ROS generation. In addition, in vivo studies further confirmed that inhibition of ER stress or oxidative stress partially abolished the butein-induced inhibition of tumor growth. Therefore, butein is a potential therapeutic agent for NSCLC, and its anticarcinogenic action might be mediated by ER stress-dependent ROS generation and the apoptosis pathway.
    Keywords:  Apoptosis; Butein; Endoplasmic reticulum stress; Non-small-cell lung cancer; Oxidative stress
    DOI:  https://doi.org/10.7150/ijbs.33790
  3. Int J Biol Sci. 2019 ;15(8): 1755-1770
    Li J, Gu Y, Zhang W, Bao CY, Li CR, Zhang JY, Liu T, Li S, Huang JX, Xie ZG, Hua SC, Wan Y.
      Diselenide-containing paclitaxel nanoparticles (SePTX NPs) indicated selectivity of cytotoxicity between cancerous and normal cells in our previous work. Herein, the mechanism is revealed by molecular biology in detail. Cancer cells and normal cells were treated with the SePTX NPs and cell proliferation was measured using 3-(4, 5-dimethyl-2-thiazolyl)-2, 5-diphenyl-2-H-tetrazolium bromide (MTT) assay and cell morphology. Measurement of reactive oxygen species (ROS) levels and biochemical parameters were employed to monitor oxidative stress of the cells. JC-1 assay was used to detect the mitochondrial dysfunction of the cells. Terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) analysis was used to detect apoptosis of the cells. Immunofluorescence analysis and western blotting were employed to monitor changes in signaling pathway-related proteins. Compared with PTX, SePTX NPs has a good selectivity to cancer cells and can obviously induce the proliferation damage of cancer cells, but has no significant toxicity to normal cells, indicating that SePTX NPs has a specific killing effect on cancer cells. The results of mechanism research show that SePTX NPs can successfully inhibit the depolymerization of microtubules and induce cell cycle arrest, which is related to the upregulation of p53 and CyclinB1. Simultaneously, SePTX NPs can successfully induce oxidative stress, cause mitochondrial dysfunction, resulting in mitochondrial pathway-mediated apoptosis, which is related to the upregulation of autophagy-related protein LC3-II. On the other hand, lewis lung cancer C57BL/6 mice were used to evaluate the anti-tumor effects of SePTX NPs in vivo. Our data show that SePTX NPs exhibited high inhibiting efficiency against the growth of tumors and were able to reduce the side effects. Collectively, these data indicate that the high antitumor effect and selective cytotoxicities of SePTX NPs is promising in future cancer therapy.
    Keywords:  Diselenide; Molecular mechanism; Nanoparticles; Paclitaxel; Selective cytotoxicity
    DOI:  https://doi.org/10.7150/ijbs.34878
  4. Int J Mol Med. 2019 Jul 23.
    Ahmad R, Vaali-Mohammed MA, Elwatidy M, Al-Obeed O, Al-Khayal K, Eldehna WM, Abdel-Aziz HA, Alafeefy A, Abdulla M.
      The emergence of colorectal cancer in developed nations can be attributed to dietary habits, smoking, a sedentary lifestyle and obesity. Several treatment regimens are available for primary and metastatic colorectal cancer; however, these treatment options have had limited impact on cure and disease‑free survival, and novel agents need to be developed for treating colorectal cancer. Thus, the objective of this study was to explore the anticancer mechanism of a benzo(1,3)dioxol‑based derivative of sulfonamide. The compound's inhibitory effect on cell proliferation was determined using the MTT assay and the xCelligence RTDP machine. Alternations in the expression of Bcl‑2 and inhibitor of apoptosis protein families were detected by western blotting. Apoptotic marker protein expression, including cytochrome c and cleaved poly(ADP‑ribose)polymerase was measured in the cytosolic extract of cells. Apoptosis and necrosis were detected by flow cytometry and immunofluorescence. Reactive oxygen species (ROS), and activation of caspase‑3 and caspase‑7 were measured using flow cytometry. Activation of the JNK pathway was detected by western blotting. We investigated the molecular mechanism of action of the sulfonamide derivative on colorectal cancer cells and found that the compound possesses a potent anticancer effect, which is primarily exerted by inducing apoptosis and necrosis. Interestingly, this compound exhibited little antiproliferative effect against the normal colonic epithelial cell line FHC. Furthermore, our results showed that the compound could significantly increase ROS production. Apoptosis induction could be attenuated by the free oxygen radical scavenger N‑acetyl cysteine (NAC), indicating that the antiproliferative effect of this compound on colorectal cancer cells is at least partially dependent on the redox balance. In addition, JNK signaling was activated by treatment with this derivative, which led to the induction of apoptosis. On the contrary, a JNK inhibitor could suppress the cell death induced by this compound. Our findings thus suggested a novel anticancer mechanism of a benzo(1,3)dioxol‑based derivative of sulfonamide for colorectal cancer cells and may have therapeutic potential for the treatment of colorectal cancer; however, further investigation is required.
    DOI:  https://doi.org/10.3892/ijmm.2019.4284
  5. Pharmacology. 2019 Jul 30. 1-10
    Chen Q, Wu S, Lu T, Chen J, Xu Z, Chen J.
      Sulforaphane (SFN) is considered an antioxidant agent, but the biological effects on hypoxia-treated osteoblasts remain unclear. Therefore, the aims of this study were to investigate the effects of SFN on the activity and mineralization of osteoblasts in hypoxia. Osteoblasts were treated with hypoxia with or without SFN, and apoptosis was assayed with caspase 3 Activity Assay Kit and flow cytometer. The levels of reactive oxygen species (ROS) were measured with DCFH-DA. The levels of glutathione (GSH) and glutathione disulphide were determined by the o-phthalaldehyde fluorimetric assay. Mineralization of Osteoblasts was detected by Alizarin red staining and alkaline phosphatase (ALP) staining, and the relative proteins levels were examined by Western blotting. Our results showed that SFN reduced the hypoxia-mediated apoptosis and ROS levels in osteoblasts. The utilization of SFN improved the inhibitory effect of osteoblast mineralization by hypoxia. Additionally, the effect of alleviating hypoxia by SFN will be an increase in osteoblast activity. These findings clarify the effects of SFN on hypoxia-treated osteogenesis and will help identify novel therapeutic strategies for the protection of skeletal health.
    Keywords:  Hypoxia; Normoxia; Osteoblast; Oxidative Stress; Sulforaphane
    DOI:  https://doi.org/10.1159/000500846
  6. Biomed Pharmacother. 2019 Jul 29. pii: S0753-3322(19)32919-1. [Epub ahead of print]118 109232
    Wang XW, Tian RM, Yang YQ, Lu ZY, Han XD, Liu XS, Mao W, Xu P, Xu HT, Liu B.
      Triptolide(T9) is a predominant bioactive component extracted from Chinese herb Tripterygium wilfordii Hook F. (TwHF), and has multiple pharmacological activities, such as immunosuppressive and anti-inflammatory activities, et al. However, severe adverse effects and toxicity, particularly nephrotoxicity, limit its clinical application. It has been demonstrated that the activation of nuclear factor erythroid 2-related factor 2 (Nrf2) signaling pathway could alleviate T9-induced nephrocyte damage. The aim of this study was to investigate the potential protective role of triptriolide (T11) against T9-induced nephrocyte apoptosis in vitro and in vivo. Renal injury models were established in human kidney 2 (HK2) cells and BALB/c mice using T9, and the protective effects of T11 were probed in vitro and in vivo, respectively. T9 induced nephrocyte damage in HK2 cells and BALB/c mice by induction of reactive oxygen species (ROS), lactate dehydrogenase (LDH), malondialdehyde (MDA) and glutathione (GSH) and reduction of superoxide dismutase (SOD), which resulted in the apoptosis of nephrocyte and injury of renal function. While, pretreatment of T11 effectively reversed these changes, resulting in the obvious decrease of oxidative stress and renal function parameters, ameliorated nephrocyte apoptosis, improved cell morphology, and higher increase of Nrf2, NAD(P)H: quinine oxidoreductase 1 (NQO1) and heme oxygenase 1 (HO-1) protein levels in vitro and in vivo. Altogether, T11 protected against T9-induced nephrocyte apoptosis possibly via suppressing oxidative stress.
    Keywords:  Apoptosis; Nrf2 pathway; Oxidative stress; Triptolide; Triptriolide
    DOI:  https://doi.org/10.1016/j.biopha.2019.109232
  7. J Appl Toxicol. 2019 Aug 01.
    Xiong G, Zhao L, Yan M, Wang X, Zhou Z, Chang X.
      The developing brain is uniquely vulnerable to toxic chemical exposures. Studies indicate that neural stem cell (NSC) self-renewal is susceptible to oxidative stress caused by xenobiotics. However, the impact of antioxidants on NSC self-renewal and the potential mechanisms remain elusive. In this study, primary murine neural progenitor cells (mNPCs) from the subventricular zone were used as a research model. In addition, paraquat (PQ) was used to elicit oxidative stress and N-acetylcysteine (NAC) was used as a powerful antioxidant. mNPCs were treated with 80 μm PQ for 24 hours with or without 4 hours of NAC pretreatment. Our results showed that PQ treatment increased intracellular reactive oxygen species production, decreased cell viability and DNA synthesis, and promoted cell apoptosis. Meanwhile, pretreatment with NAC alleviated PQ-induced cytotoxicity in mNPCs. To elucidate the mechanisms further, we found that NAC pretreatment prevented PQ-induced reactive oxygen species production, mitochondrial fragmentation and autophagy in mNPCs. NAC-pretreated cells showed increased anti-apoptotic protein Bcl-2 and decreased pro-apoptotic protein Bax expression. Similarly, NAC pretreatment increased p-mTOR and decreased LC3B-II protein expression. Moreover, NAC decreased mitophagy related mRNA Pink1 and Parkin expression. Taken together, our results suggested that the antioxidant NAC treatment significantly attenuated PQ-induced mNPC self-renewal disruption through decreasing autophagy and salvaging mitochondrial morphology. These findings revealed a potential mechanism for neurological treatment relating to antioxidant and suggested potentially relevant implications for PQ-related neurodegenerative disorders. Thus, our study also provided insight into therapeutic strategies for the neurotoxic effects of oxidative stress-associated toxicants.
    Keywords:  N-acetylcysteine; apoptosis; autophagy; mitochondrial morphology; neural progenitor cells; paraquat
    DOI:  https://doi.org/10.1002/jat.3839
  8. Chemosphere. 2019 Jul 23. pii: S0045-6535(19)31656-X. [Epub ahead of print]237 124435
    Zhang JW, Xu DQ, Feng XZ.
      Glyphosate is a high-efficiency, low-toxicity, broad-spectrum herbicide. The residues of glyphosate-based herbicides are frequent pollutants in the environment. However, the effects of glyphosate on oocyte maturation, as well as its possible mechanisms, remain unclear. The present study revealed that mouse oocytes had reduced rates of germinal vesicle breakdown (GVBD) and first polar body extrusion (PBE) after treatment with 500 μM glyphosate. Reactive oxygen species (ROS) were found in mouse oocytes exposed to glyphosate, as shown by changes in the mRNA expression of related antioxidant enzyme genes (cat, sod2, gpx). After 14 h of exposure to glyphosate, metaphase II (MII) mouse oocytes displayed an abnormal spindle morphology and DNA double-strand breaks (DNA-DSBs). Simultaneously, mitochondria showed an aggregated distribution and decreased membrane potential in mouse oocytes exposed to glyphosate. The protein expression levels of apoptosis factors (Bax, Bcl-2) and the mRNA expression levels of apoptosis-related genes (bax, bcl-2, caspase3) were measured by Western blot and qRT-PCR, respectively. Meanwhile, the expression levels of autophagy-related genes (lc3, atg14, mtor) and proteins (LC3, Atg12) were significantly decreased in the glyphosate treatment group compared with the control group. Collectively, our results indicated that glyphosate exposure could interfere with mouse oocyte maturation by generating oxidative stress and early apoptosis.
    Keywords:  Apoptosis; Autophagy; Glyphosate; Oocyte quality; Oxidative stress
    DOI:  https://doi.org/10.1016/j.chemosphere.2019.124435
  9. Front Pharmacol. 2019 ;10 762
    Liu Y, Shao E, Zhang Z, Yang D, Li G, Cao H, Huang H.
      Indolizine derivatives are a class of compounds with excellent biological activity. In this study, a series of indolizine derivatives, compound 1 (C1), compound 2 (C2), compound 3 (C3), and compound 4 (C4), were synthesized. 3-(4,5-dimethylthiazole)-2,5-diphenyltetraazolium bromide (MTT) assay was used to evaluate their cytotoxicity against HepG2 (p53-wild), A549, and HeLa cell lines. HepG2 cells apoptosis induced by C3 was determined using Hoechst staining and acridine orange/ethidium bromide staining. Cells' apoptotic ratio was measured by Annexin V-FITC/PI double staining. Changes in mitochondrial membrane potential and intracellular reactive oxygen species (ROS) in HepG2 cells after C3 treatment were determined. Immunofluorescence staining and Western blot analysis were carried out to detect p53 levels and analyze the apoptosis-associated proteins, respectively. Moreover, the cytotoxic activity of C3 was examined in two other hepatocellular carcinoma (HCC) cell lines with different p53 status including Huh-7 cells (p53-mutant) and Hep3B cells (p53-null). The results indicated that C3 showed stronger inhibition towards HepG2 cells than other cell lines. Fluorescent staining and flow cytometry analysis confirmed that C3 induced apoptosis of HepG2 cells. C3 could also increase intracellular ROS and cause a decrease in the mitochondrial membrane potential. C3 promoted p53 activation and increased p53 accumulation in nuclei. The expression of p53 and Bax was increased with the down-regulation of Bcl-2, which promoted the release of cytochrome c and caspase-3 activation. Collectively, the study demonstrated that C3 caused HepG2 cell apoptosis via the mitochondria p53 pathway. These results inspired us to further develop indolizine derivatives as potential potent inhibitors against liver cancer.
    Keywords:  HepG2 cells; apoptosis; indolizine derivatives; mitochondrion; p53
    DOI:  https://doi.org/10.3389/fphar.2019.00762
  10. Oxid Med Cell Longev. 2019 ;2019 8506195
    Yan T, Huang J, Nisar MF, Wan C, Huang W.
      Drug-induced liver injury (DILI) is a major cause of acute liver failure (ALF) as a result of accumulated drugs in the human body metabolized into toxic agents and helps generate heavy oxidative stress, inflammation, and apoptosis, which induces necrosis in hepatocytes and ultimately damages the liver. Sirtuin 1 (SIRT1) is said to have multiple vital roles in cell proliferation, aging, and antistress systems of the human body. The levels of SIRT1 and its activation precisely modulate its critical role in the interaction between multiple step procedures of DILI. The nuclear factor kappa-light-chain-enhancer of activated B cell- (NF-κB-) mediated inflammation signaling pathway, reactive oxygen species (ROS), DNA damage, mitochondrial membrane potential collapse, and endoplasmic reticulum (ER) stress also contribute to aggravate DILI. Apoptosis is regarded as the terminal reaction followed by multiple signaling cascades including caspases, p53, and mitochondrial dysfunction which have been said to contribute in DILI. The SIRT1 activator is regarded as a potential candidate for DILI, because the former could inhibit signaling of p53, NF-κB, and ER stress. On the other hand, overexpression of SIRT1 also enhances the activation of antioxidant responses via Kelch-like ECH-associated protein 1- (Keap1-) nuclear factor- (erythroid-derived 2-) like 2 (Nrf2) signaling. The current manuscript will highlight the mechanism of DILI and the interaction of SIRT1 with various cytoplasmic factors leading to DILI along with the summary of potent SIRT1 agonists.
    DOI:  https://doi.org/10.1155/2019/8506195
  11. Biol Trace Elem Res. 2019 Aug 01.
    Kar F, Hacioglu C, Senturk H, Donmez DB, Kanbak G.
      Ischemia/reperfusion (I/R) injury is associated with a strong inflammatory and oxidative stress response to hypoxia and reperfusion that impair organ function. We aimed to investigate the role of oxidative stress, renal inflammation, and apoptosis in the injury of the kidney tissue after ischemic reperfusion, and the protective effect of dose-dependent boric acid administration. For this purpose, 35 Sprague Dawley albino rats were divided into five groups of seven animals in each group: Sham, I/R and I/R + boric acid (BA) (i.p at doses of 50, 100, and 200 mg/kg). All animals underwent nephrectomy (the right kidney was removed) and were expected to recover for 15 days. After recovery, each animal received 45 min of ischemia. BA was injected intraperitoneally 10 min before reperfusion and a 24-h reperfusion procedure was performed. Sham group only underwent surgical stress procedure. In order to investigate the oxidative stress induced by I/R injury and antioxidant effects of different BA doses in the kidney tissue, TAS, TOS, MDA, SOD, CAT, and GSH levels were measured. DNA fragmentation, cytochrome C levels, caspase 3 activity were measured to determine apoptotic index in tissue. IL-6 and TNF-α levels were measured in the evaluation of inflammation. Hematoxylin-eosin and TUNEL staining was performed for histopathological examinations. As a result, increased oxidative stress, inflammation, and apoptosis after I/R were decreased with different doses of BA treatment. The application of high-dose BA was found to be lower in anti-apoptotic, anti-inflammatory, and antioxidant effects than in the low-dose groups.
    Keywords:  Apoptosis; Boric acid; Inflammation; Ischemia/reperfusion; Kidney; Oxidative stress
    DOI:  https://doi.org/10.1007/s12011-019-01824-1
  12. Cell Stress Chaperones. 2019 Jul 31.
    Kar F, Hacioglu C, Kacar S, Sahinturk V, Kanbak G.
      Prostate cancer is the main cause of cancer-related mortality in men around the world and an important health problem. DU-145 human prostate cancer cells provide an opportunity to investigate prostate cancer. Betaine has a number of anticancer effects, such as inactivation of carcinogens, inhibition of cancer cell proliferation, angiogenesis, and metastasis. However, there is no study investigating the effects of betaine on DU-145 cells. The aim of this study was to evaluate the effects of different concentrations of betaine on the oxidative stress, apoptosis, and inflammation on DU-145 cells. Firstly, we proved the cytotoxic activity of betaine (0 to 150 mg/ml) on DU-145 cells by using 3-(4, 5-dimethylthiazol, 2-yl)-2, 5-diphenyl tetrazolium bromide (MTT) and defined the optimal concentration of betaine. Then, by employing the doses found in MTT, the levels of antioxidant (GSH, SOD, CAT, and TAS) and oxidant (MDA and TOS) molecules, pro-inflammatory cytokines (TNF-a and IL-6), apoptotic proteins (CYCS and CASP3), and DNA fragmentation were measured. Morphological changes and apoptosis were evaluated using H&E technique, Bax and Bcl-2 immunohistochemistry. Results suggested that betaine caused oxidative stress, inflammation, inhibition of cell growth, apoptosis, and morphological alterations in DU-145 cells dose-dependently. Furthermore, treatments with increasing betaine concentrations decreased the antioxidant levels in cells. We actually revealed that betaine, known as an antioxidant, may prevent cell proliferation by acting as an oxidant in certain doses. In conclusion, betaine may act as a biological response modifier in prostate cancer treatment in a dose-dependent manner.
    Keywords:  Apoptosis; Betaine; Cell morphology; DU-145 prostate cancer cell; Oxidative stress
    DOI:  https://doi.org/10.1007/s12192-019-01022-x
  13. Anticancer Agents Med Chem. 2019 Jul 31.
    Khan F, Singh VK, Saeed M, Kausar MA, Ansari IA.
      BACKGROUND: Several studies have revealed that abnormal activation of Notch signaling is closely related with the development and progression of prostate cancer. Although there are numerous therapeutic strategies, a more effective modality with least side effects is urgently required for the treatment of prostate cancer. Carvacrol is a monoterpenoid phenol and majorly present in the essential oils of Lamiaceae family plants. Many previous reports have shown various biological activities of carvacrol like antioxidant, anti-inflammatory and anticancer properties. Recently, we have shown potent anticancer property of carvacrol against prostate cancer cell line DU145. In the current study, we report the chemopreventive and therapeutic potential of carvacrol against another prostate cancer cell line PC-3 with its detailed mechanism of action.METHODS: To determine the effect of the carvacrol on prostate cancer cells, the cell viability was estimated by MTT assay and cell death was estimated by LDH release assay. The apoptotic assay was performed by DAPI staining and FITC-Annexin V assay. Reactive Oxygen Species (ROS) was estimated by DCFDA method. Cell cycle analysis was performed by flow cytometry. Gene expression analysis was performed quantitative real time PCR.
    RESULTS: Our results suggested that the carvacrol treatment significantly reduced the cell viability of PC-3 cells in a dose- and time-dependent manner. The antiproliferative action of carvacrol was correlated with apoptosis which was confirmed by nuclear condensation, FITC-Annexin V assay, modulation in expression of Bax, Bcl-2 and caspase activation. The mechanistic insight into carvacrol-induced apoptosis leads to finding of elevated level of Reactive Oxygen Species (ROS) and mitochondrial membrane potential disruption. Cell cycle analysis revealed that carvacrol prevented cell cycle in G0/G1 that was associated with decline in expression of cyclin D1 and Cyclin-Dependent Kinase 4 (CDK4) and augmented expression of CDK inhibitor p21. Having been said the role of hyperactivation of Notch signaling in prostate cancer, we also deciphered that carvacrol could inhibit Notch signaling in PC-3 cells via downregulation of Notch-1, and Jagged-1.
    CONCLUSION: Thus, our previous and current findings have established the strong potential of carvacrol as a chemopreventive agent against androgen-independent human prostate cancer cells.
    Keywords:  Jagged-1; PC-3; Prostate cancer; apoptosis; cell cycle; notch
    DOI:  https://doi.org/10.2174/1871520619666190731152942
  14. Mar Drugs. 2019 Jul 27. pii: E444. [Epub ahead of print]17(8):
    Zhen AX, Hyun YJ, Piao MJ, Fernando PDSM, Kang KA, Ahn MJ, Yi JM, Kang HK, Koh YS, Lee NH, Hyun JW.
      Toxicity of particulate matter (PM) towards the epidermis has been well established in many epidemiological studies. It is manifested in cancer, aging, and skin damage. In this study, we aimed to show the mechanism underlying the protective effects of eckol, a phlorotannin isolated from brown seaweed, on human HaCaT keratinocytes against PM2.5-induced cell damage. First, to elucidate the underlying mechanism of toxicity of PM2.5, we checked the reactive oxygen species (ROS) level, which contributed significantly to cell damage. Experimental data indicate that excessive ROS caused damage to lipids, proteins, and DNA and induced mitochondrial dysfunction. Furthermore, eckol (30 μM) decreased ROS generation, ensuring the stability of molecules, and maintaining a steady mitochondrial state. The western blot analysis showed that PM2.5 promoted apoptosis-related protein levels and activated MAPK signaling pathway, whereas eckol protected cells from apoptosis by inhibiting MAPK signaling pathway. This was further reinforced by detailed investigations using MAPK inhibitors. Thus, our results demonstrated that inhibition of PM2.5-induced cell apoptosis by eckol was through MAPK signaling pathway. In conclusion, eckol could protect skin HaCaT cells from PM2.5-induced apoptosis via inhibiting ROS generation.
    Keywords:  keratinocytes; particulate matter; phlorotannin; reactive oxygen species
    DOI:  https://doi.org/10.3390/md17080444
  15. J Recept Signal Transduct Res. 2019 Apr;39(2): 134-145
    Xie Z, Zhou Y, Duan X, Yang L.
      Context: Epidermal cells play an important role in regulating the regeneration of skin after burns and wounds. Objective: The aim of our study is to explore the role of Tanshinone IIA (Tan IIA) in the apoptosis of epidermal HaCaT cells induced by H2O2, with a focus on mitochondrial homeostasis and inverted formin-2 (INF2). Materials and methods: Cellular viability was determined using the MTT assay, TUNEL staining, western blot analysis and LDH release assay. Adenovirus-loaded INF2 was transfected into HaCaT cells to overexpress INF2 in the presence of Tan IIA treatment. Mitochondrial function was determined using JC-1 staining, mitochondrial ROS staining, immunofluorescence and western blotting. Results: Oxidative stress promoted the death of HaCaT cells and this effect could be reversed by Tan IIA. At the molecular levels, Tan IIA treatment sustained mitochondrial energy metabolism, repressed mitochondrial ROS generation, stabilized mitochondrial potential, and blocked the mitochondrial apoptotic pathway. Furthermore, we demonstrated that Tan IIA modulated mitochondrial homeostasis via affecting INF2-related mitochondrial stress. Overexpression of INF2 could abolish the protective effects of Tan IIA on HaCaT cells viability and mitochondrial function. Besides, we also reported that Tan IIA regulated INF2 expression via the ERK pathway; inhibition of this pathway abrogated the beneficial effects of Tan IIA on HaCaT cells survival and mitochondrial homeostasis. Conclusions: Overall, our results indicated that oxidative stress-mediated HaCaT cells apoptosis could be reversed by Tan IIA treatment via reducing INF2-related mitochondrial stress in a manner dependent on the ERK signaling pathway.
    Keywords:  ERK signaling pathway; HaCaT cells; INF2; Tan IIA; mitochondrial stress
    DOI:  https://doi.org/10.1080/10799893.2019.1638402
  16. Biomed Res Int. 2019 ;2019 5962014
    Fan Z, Wang X, Zhang M, Zhao C, Mei C, Li P.
      Background: Oxidative stress due to reactive oxygen species plays a central role in pathophysiology of neurodegenerative diseases. Inhibition of mitogen-activated protein kinase (MAPK) cascades attenuates the oxidative induced cell stress and behaves as potential neuroprotection agent.Materials and Methods: In this study, we evaluate hydrogen peroxide induced neural cell stress and determine how different MAPK inhibitors restore the cell damage.
    Results: The results indicated that oxidative stress induced by neural cell damage commonly exists, and MAPK inhibitors partially and selectively attenuated the cell damage by reducing ROS production and cell apoptosis. The cultured neurons are more susceptible to hydrogen peroxide than subculture cells.
    Conclusion: We conclude that the essential role of different MAPK inhibitors is to attenuate the hydrogen peroxide induced neuronal cell damage. Those data broaden the implication between individual neural cells and different MAPK inhibitors and give clues for oxidative stress induced neural diseases.
    DOI:  https://doi.org/10.1155/2019/5962014
  17. J Food Biochem. 2019 Feb;43(2): e12708
    Liu S, Sui Q, Zou J, Zhao Y, Chang X.
      This study investigated the effect of a hawthorn polyphenol extract (HPE) on ultraviolet B (UVB)-induced damage in HaCaT cells and mice. High-performance liquid chromatography/electrospray ionization tandem mass spectrometry was used to analyze the phenolic composition of HPE. The protective effects of HPE and its main components were compared in HaCaT cells. An enzyme-linked immunosorbent assay was used to detect DNA damage (8-hydroxydeoxyguanosine levels). Flow cytometry and western blotting were used to measure the extent of apoptosis and the levels of apoptosis-related proteins, respectively. Treatment with HPE or its polyphenol components inhibited the UVB-induced damage by removing an excess of reactive oxygen species (ROS), reducing DNA damage and p53 activation, regulating the protein expression of B-cell lymphoma 2 family members toward antiapoptotic ratios, and reducing caspase activation. Similar effects were observed in a UVB-irradiated mouse skin, as detected using terminal deoxynucleotidyl transferase dUTP nick-end labeling, immunohistochemistry, and western blotting assays. These results suggest that HPE can be used as a natural dietary supplement for the prevention and treatment of UVB radiation-induced skin damage. PRACTICAL APPLICATIONS: Hawthorn (Crataegus pinnatifida) shows antioxidant, anti-inflammatory, and lipid-lowering effects. As natural, healthy, and effective additives, HPEs have been widely used in food and health products. The results of this study reveal the molecular mechanisms underlying HPE effects, showing that HPE reverses the effects of UVB irradiation via removal of an excess of ROS and reduction of DNA damage and p53 expression in vitro and in vivo. Consequently, HPE upregulates the expression of antiapoptotic BCL-2 and downregulates that of proapoptotic BAX, thereby reducing the activation of caspase-3/9 and inhibiting apoptosis. These findings suggest that HPE can be used as the base ingredient for antiphotoaging food products. This study provides both theoretical and experimental background for hawthorn deep processing and utilization.
    Keywords:  HaCaT cell; ROS; UVB damage; immunohistochemistry; mitochondrial apoptosis pathway
    DOI:  https://doi.org/10.1111/jfbc.12708
  18. J Cardiovasc Pharmacol. 2019 Jul 22.
    Yan R, Yan J, Chen X, Yu Y, Sun T.
      OBJECTIVE: Atherosclerosis (AS) contributes to the development of several cardiovascular diseases such as myocardial infarction and stroke. Oxidized low-density lipoprotein (Ox-LDL)-induced endothelial cells injury plays a key role in the pathogenesis of AS. Thus, this study was carried out to examine the effects of a naturally occurring flavonoid compound, xanthoangelol (XAG), on ox-LDL-induced cell injury.MATERIALS AND METHODS: Human umbilical vein endothelial cells (HUVECs) were used as in vitro cell model. The number of viable cells was determined using CCK-8 assay. Cell apoptosis was detected using Hoechest staining. Percentage of apoptotic cells was quantified by flow cytometry. The cellular levels of malondialdehyde (MDA), superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GSH-Px) were determined using enzyme linked immune-sorbent assays (ELISA). Cellular reactive oxygen species (ROS) level was detected by flow cytometry after florescence staining. The mRNA expression levels of nuclear factor E2 related factor-2(Nrf2), Heme Oxygenase-1(HO-1) and NQO-1 were determined using qRT-PCR assay. The protein levels of cleaved caspase-3, cleaved PARP, Bax, Bcl-2, Nrf2, Keap1, HO-1 and NQO-1 were measured via western blot assay. The HUVECs were transfected with Nrf2 siRNA to reduce the expression of Nrf2.
    RESULTS: XAG could effectively protect against ox-LDL-stimulated cell death in HUVECs. These cytoprotective effects were due to its anti-apoptotic and anti-oxidant activities. As supported by the increase of SOD, CAT, and GSH-Px activities, and the decrease of MDA and ROS levels in injured HUVECs induced by ox-LDL. Moreover, the results showed that XAG activated Nrf2/ARE signaling in a dose-dependent manner. Importantly, blockade of Nrf2 signaling using siRNA or specific inhibitor notably abolished the cytoprotective activities of XAG.
    CONCLUSIONS: These data suggests that XAG cytoprotects against ox-LDL-induced cell injury through activating Nrf2/ARE-mediated anti-oxidative stress. Cumulatively, these findings show that EX has the potential to prevent and treat AS.
    DOI:  https://doi.org/10.1097/FJC.0000000000000699
  19. Zhonghua Gan Zang Bing Za Zhi. 2019 Jun 20. 27(6): 445-449
    Ao N, Yang J, Du J.
      Objective: To observe whether liraglutide protects HepG2 cells from lipotoxicity by affecting mitogen-activated protein kinase (MAPKs) pathway. Methods: HepG2 cells were induced with 400μmol/L palmitic acid, and cells were treated with a final concentration of 100 nmol/L liraglutide. In addition, JNK inhibitor (SP600125) and p38 MAPK inhibitor (SB203580) were added in advance, respectively. Apoptosis rate, malondialdehyde (MDA) content, and caspase3 activity were detected. Western blot was used to detect p38 mitogen-activated protein kinase (p38 MAPK), c-jun amino terminal kinase (JNK), cytochrome oxidase P450 2E1 (CYP2E1), glucose regulatory protein 78 (GRP78), activated caspase 3, B cell lymphoma associated Protein X (Bax), B cell lymphoma 2 (Bcl-2), and expression of C/EBP homologous protein (CHOP) protein. LSD or Dunnett's T3 test were used to compare the mean of multiple samples. Results: Palmitic acid increased the phosphorylation of p38 MAPK and JNK in HepG2 cells (P< 0.05). Furthermore, it increased the expression of GRP78, CHOP, CYP2E1, MDA, Bax, caspase3 and apoptosis rate, but inhibited the expression of Bcl-2 (Pvalue < 0.05). SP600125 and SB203580 had inhibited oxidative stress and apoptosis induced by palmitic acid (including CYP2E1, MDA, Bax, Bcl-2, caspase3, CHOP) (P< 0.05). The phosphorylation level of p38 MAPK and JNK was reduced with liraglutide and the expression of apoptosis-related proteins (Bax, Bcl-2, caspase3, CHOP) (P< 0.05) was regulated. There was no significant difference in the effect of liraglutide on apoptotic proteins (Bax, Bcl-2, caspase-3, CHOP) (P> 0.05) after pretreatment with those two inhibitors. Conclusion: Palmitic acid has strong lipotoxicity to HepG2 cells and induces apoptosis. Glucagon-like peptide-1 analogue, liraglutide may improve lipotoxicity of palmitic acid by mediating p38 MAPK and JNK pathways.
    Keywords:  Apoptosis; Endoplasmic reticulum stress; Liraglutide; Oxidative stress
    DOI:  https://doi.org/10.3760/cma.j.issn.1007-3418.2019.06.011
  20. Evid Based Complement Alternat Med. 2019 ;2019 8450158
    Mbaveng AT, Damen F, Simo Mpetga JD, Awouafack MD, Tane P, Kuete V, Efferth T.
      The effectiveness of anticancer chemotherapy is greatly impeded by the resistance of malignant cells to cytotoxic drugs. In this study, the cytotoxicity of the crude extract (DCB) and compounds isolated from the bark of Dichrostachys cinerea, namely, betulinic acid (1), glyceryl-1-hexacosanoate (2), 7-hydroxy-2-(4-hydroxyphenyl)-4H-chromen-4-one (3), and 6-hydroxy-2-(4-hydroxyphenyl)-4H-chromen-4-one (4), was investigated. The study was extended to the assessment of the mode of induction of apoptosis by DCB and compound 1. The resazurin reduction assay was used for cytotoxicity studies. Assessments of cell cycle distribution, apoptosis, mitochondrial membrane potential (MMP), and reactive oxygen species (ROS) were performed by flow cytometry. Constituents of DCB were isolated by column chromatography. Triterpenoid 1 and flavone 4 had cytotoxic effects towards the 9 tested cancer cell lines with IC50 values below 50 μM. The recorded IC50 values varied from 7.65 μM (towards multidrug-resistant CEM-ADR5000 leukemia cells) to 44.17 μM (against HepG2 hepatocarcinoma cells) for 1, 18.90 μM (CCRF-CEM leukemia cells) to 88.86 μM (against HCT116p53+/+ colon adenocarcinoma cells) for 4, and 0.02 μM (against CCRF-CEM cells) to 122.96 μM (against CEM/ADR5000 cells) for doxorubicin. DCB induced apoptosis in CCRF-CEM cells mostly mediated by MMP alteration and enhanced ROS production; compound 1 induced apoptosis through caspases activation and MMP alteration and increased ROS production. Dichrostachys cinerea is an interesting cytotoxic plant and deserves more studies leading to new antineoplastic agents to fight cancer and mostly leukemia.
    DOI:  https://doi.org/10.1155/2019/8450158
  21. Can J Physiol Pharmacol. 2019 Jul 31. 1-5
    Sun Y, Nemec-Bakk AS, Mallik AU, Bagchi AK, Singal PK, Khaper N.
      The objective of this study was to analyze the cardioprotective roles of 3 wild blueberry genotypes and one commercial blueberry genotype by measuring markers of oxidative stress and cell death in H9c2 cardiac cells exposed to doxorubicin. Ripe berries of the 3 wild blueberry genotypes were collected from a 10-year-old clearcut forest near Nipigon, Ontario, Canada (49°1'39″N, 87°52'21″W), whereas the commercial blueberries were purchased from a local grocery store. H9c2 cardiac cells were incubated with 15 μg gallic acid equivalent/mL blueberry extract for 4 h followed by 5 μM doxorubicin for 4 h, and oxidative stress and active caspase 3/7 were analyzed. The surface area as well as total phenolic content was significantly higher in all 3 wild blueberry genotypes compared with the commercial species. Increase in oxidative stress due to doxorubicin exposure was attenuated by pre-treatment with all 3 types of wild blueberries but not by commercial berries. Furthermore, increase in caspase 3/7 activity was also attenuated by all 3 wild genotypes as well. These data demonstrate that wild blueberry extracts can attenuate doxorubicin-induced damage to H9c2 cardiomyocytes through reduction in oxidative stress and apoptosis, whereas the commercial blueberry had little effect.
    Keywords:  H9c2 cardiac cells; antioxidant; antioxydant; apoptose; apoptosis; blueberries; cellules cardiaques H9c2; doxorubicin; doxorubicine; myrtilles; oxidative stress; phenols; phénols; stress oxydatif
    DOI:  https://doi.org/10.1139/cjpp-2019-0031
  22. Molecules. 2019 Jul 26. pii: E2727. [Epub ahead of print]24(15):
    Mo C, Shetti D, Wei K.
      Psoriasis is a recurrent skin disease described as keratinocyte hyperproliferation and aberrant differentiation. Erianin, a bibenzyl compound extracted from Dendrobium chrysotoxum, has displayed antitumor and anti-angiogenesis effects. However, the effects of erianin on a human keratinocyte cell line (HaCaT) are not fully understood. In the present study, we explored the effect of erianin on proliferation and apoptosis in HaCaT cells. Our results indicated that treatment with erianin ranging from 12.5 nM to 50 nM inhibited proliferation and induced apoptosis of HaCaT cells. In addition, erianin-induced apoptosis was accompanied by elevated reactive oxygen species (ROS). The ROS scavenger N-acetyl-cysteine (NAC) attenuated this elevation. Moreover, treatment with erianin induced activation of the c-Jun N-terminal kinase (JNK)/c-Jun signaling pathway and suppressed the protein kinase B (AKT)/mammalian target of rapamycin (mTOR) signaling pathway, while pretreatment with NAC also reversed these effects. Collectively, these data demonstrated that erianin inhibited proliferation and induced apoptosis of HaCaT cells through ROS-mediated JNK/c-Jun and AKT/mTOR signaling pathways. Erianin could be recognized as a potential anti-psoriasis drug.
    Keywords:  AKT/mTOR; JNK/c-Jun; ROS; apoptosis; erianin; keratinocyte; psoriasis
    DOI:  https://doi.org/10.3390/molecules24152727
  23. J Infect Chemother. 2019 Jul 25. pii: S1341-321X(19)30206-5. [Epub ahead of print]
    Azab SS, Kamel I, Ismail NN, El Din Hosni H, El Fatah MA.
      INTRODUCTION: Cisplatin (CIS), which is used as a therapeutic antineoplastic agent may produce gonadotoxicity in a process linked to chemotherapy. Taurine, through its potential antioxidant effect, has a protective role against cisplatin-induced oxidative stress and apoptosis.OBJECTIVE: To investigate whether taurine intake can ameliorate testicular damage induced by cisplatin and to study the possible mechanism that mediates this action, either through its antioxidant action alone or in addition to its anti-apoptotic effects.
    PATIENTS AND METHODS: Fifty healthy adult white male albino rats were randomly distributed into five groups, each involving ten animals. The first group represents the negative control group. The other four groups received three equal doses (3 mg/kg body weight) intraperitoneal injections of cisplatin on alternate days. In the positive control group (group 2), saline only was given. Groups 3, 4 &5 received taurine in distilled water at oral doses of 50, 150, 250 mg/kg, respectively, on alternate days followed by cisplatin (each injection of cisplatin was given 1 day after taurine). On the 28th day after the first dose of normal saline, cisplatin or taurine, blood samples were examined for testosterone levels. All rats were killed and their testes were examined.
    RESULTS: Rats treated with cisplatin alone showed reduced body weight in addition to reduced testicular weight, impaired sperm counts, and oxidative stress (reduced GSH, increased MDA level), decreased plasma testosterone, apoptotic marker (increased Bax, decreased bcl2). However following taurine induction, the figures for GSH and MDA changed significantly (P < 0.005) referring to the effect of taurine as a potent antioxidant.
    CONCLUSIONS: Cisplatin-induced germ cell apoptosis may result in decreasing spermatogenesis. However, taurine could effectively prevent nearly all of these cisplatin-induced testicular abnormalities, thereby proving to be an effective cytoprotectant.
    Keywords:  Apoptosis; Cisplatin; Taurine; Testicular; oxidative stress
    DOI:  https://doi.org/10.1016/j.jiac.2019.07.004
  24. Asian Pac J Cancer Prev. 2019 07 01. pii: 88668. [Epub ahead of print]20(7): 2225-2238
    Abd-Rabou AA, Ahmed HH.
      Non-small cell lung cancer (NSCLC) and hepatocellular carcinoma (HCC) are very common in certain population around the world. Despite the recent advances in their diagnosis and therapy, their prognosis remains poor due to the development resistance to drug. Although doxorubicin (DOX) is considered to be one of the most anti-solid tumor drugs, developed resistance is contributing to unsuccessful outcome. The rationale of the current study is to explore the sensitizing capability of the DOX-treated cancer cells using the anticancer agents; bevacizumab (avastin; AV) and CCR2 inhibitor (CR) in their free- and nano-formulations. Here, the average size, polydispersity index (PDI), zeta potential, and entrpment effeciency (EE%) of the synthesized nanoparticles were measured. We investigated the effect of these platforms on the proliferation, apoptosis, necrosis, nitric oxide (NO), malondialdehyde (MDA), and zinc levels of human HCC (HepG2 and Huh-7) and NSCLC (A549) cancer cell lines. Glucose consumption rates using Huh-7 and A549 cancer cells were tested upon treatments. We demonstrated that AV and CR nano-treatments significantly suppressed A549 cell viability and activated apoptosis by NO level elevation. We concluded that AVCR NP plus DOX significantly induces A549 cytotoxicity-mediated apoptosis more than Huh-7 and HepG2 cells. This drug-drug nano-combination induced Huh-7 cytotoxicity-mediated apoptosis more than HepG2 cells. In conclusion, AVCR NP sensitized DOX-treated A549 and Huh-7 cells through reactive oxygen species (ROS)-stimulated apoptosis. Taken together, our data suggested that the CR plus AV nano-platforms would be a potential personalized medicine-based strategy for treating CCR2-positive NSCLC and HCC patients in the near future.
    Keywords:  Bevacizumab (avastin); CCR2 antagonist; Cytotoxicity; Hepatocellular carcinoma; non-small cell lung cancer
    DOI:  https://doi.org/10.31557/APJCP.2019.20.7.2225
  25. Biol Chem. 2019 Jul 29. pii: /j/bchm.ahead-of-print/hsz-2019-0160/hsz-2019-0160.xml. [Epub ahead of print]
    Ehrenfeld V, Fulda S.
      Imbalances in redox homeostasis have been described to be involved in the development, progression and relapse of leukemia. As the thioredoxin (Trx) system, one of the major cellular antioxidant networks, has been implicated in acute lymphoblastic leukemia (ALL), we investigated the therapeutic potential of Trx inhibition in ALL. Here, we show that the Trx inhibitor PX-12 reduced cell viability and induced cell death in a dose- and time-dependent manner in different ALL cell lines. This antileukemic activity was accompanied by an increase in reactive oxygen species (ROS) levels and enhanced PRDX3 dimerization. Pre-treatment with the thiol-containing ROS scavenger N-acetylcysteine (NAC), but not with non-thiol-containing scavengers α-tocopherol (α-Toc) or Mn(III)tetrakis(4-benzoic acid) porphyrin chloride (MnTBAP), significantly rescued PX-12-induced cell death. Furthermore, PX-12 triggered activation of BAK. Importantly, knockdown of BAK reduced PX-12-stimulated ROS production and cell death. Similarly, silencing of NOXA provided significant protection from PX-12-mediated cell death. The relevance of mitochondria-mediated, caspase-dependent apoptosis was further supported by data showing that PX-12 triggered cleavage of caspase-3 and that addition of the broad-range caspase inhibitor carbobenzoxy-valyl-alanyl-aspartyl-[O-methyl]-fluoromethylketone (zVAD.fmk) potently blocked cell death upon PX-12 treatment. This study provides novel insights into the mechanisms of PX-12-induced cell death in ALL and further highlights the therapeutic potential of redox-active compounds in ALL.
    Keywords:  ALL; PX-12; apoptosis; mitochondria; reactive oxygen species; thioredoxin system
    DOI:  https://doi.org/10.1515/hsz-2019-0160
  26. Life Sci. 2019 Jul 29. pii: S0024-3205(19)30631-9. [Epub ahead of print] 116704
    Taskin E, Guven C, Kaya ST, Sahin L, Kocahan S, Degirmencioglu AZ, Gur FM, Sevgiler Y.
      AIMS: Doxorubicin, an anticancer drug, has a toxic effect on many tissues such as heart, pancreas, liver, kidney, and testis. The aim of current study is to investigate whether melatonin would be protective in doxorubicin-induced beta (β) cell toxicity via HMGB1/TLR2/TLR4/MAPK/NF-қB signaling pathway.MAIN METHODS: Human pancreatic β cell (1.1B4) was used in the present study. Four experimental groups were created as control, melatonin (10 μM), doxorubicin (2 μM) and the combination of melatonin with doxorubicin. Following 24-h treatment, Mitogen-activated protein kinase (MAPKs), Toll like receptors (TLRs) including TLR2 and TLR4, pro-and anti-apoptotic protein expression levels were determined by western blotting. Total antioxidant (TAS), oxidant status (TOS) and oxidative stress index (OSI) of the cells as well as superoxide dismutase (SOD) levels were determined. Active caspase-8 activity was measured and TUNEL staining was performed to study apoptotic pathways. Mitochondrial membrane potential (MMP), some protein expressions and F-actin distribution were analyzed.
    KEY FINDINGS: Doxorubicin caused to depolarize MMP, resulting in enhancing apoptosis by activation of caspase-8 via MAPKs/NF-кB pathway via elevation of TOS and decreasing TAS. Also, doxorubicin destroyed F-actin distribution and elevated TLR2 and some apoptotic proteins, including Bax. However, co-treatment of melatonin with doxorubicin could reverse depolarization of MMP and inhibition of apoptosis through MAPK/NF-кB signaling by decreasing TOS and increasing TAS. The co-treatment reversed the alternations of TLR2, TLR4, MAPKs and apoptotic protein expressions induced by doxorubicin.
    SIGNIFICANCE: Melatonin could be a good candidate against pancreatic β cell toxicity-induced by doxorubicin through TLR2/TLR4/MAPK/NF-кB pathways.
    Keywords:  Beta cell; Doxorubicin; Melatonin; Oxidative stress; Protein kinases; Toll like receptors
    DOI:  https://doi.org/10.1016/j.lfs.2019.116704
  27. Int J Mol Med. 2019 Jul 22.
    Li X, Meng F, Li H, Hua X, Wu L, Yuan X.
      L‑carnitine (LC) is well known for its antioxidative properties. The present study aimed to evaluate the effects of LC on human lens epithelial cells (HLECs) and to analyze its regulatory mechanism in cataractogenesis. HLE B‑3 cells were cultured with hydrogen peroxide (H2O2) and were pretreated with or without LC. The Cell Counting kit‑8 assay was used to determine cell viability. Reactive oxygen species (ROS) assay kit was used to measure the cellular ROS production induced by H2O2 and LC. In addition, reverse transcription‑quantitative PCR and western blot analysis were performed to detect the expression levels of oxidative damage markers and antioxidant enzymes. Notably, ROS overproduction was observed upon exposure to H2O2, whereas LC supplementation markedly decreased ROS levels through activation of the antioxidant enzymes forkhead box O1, peroxiredoxin 4 and catalase. Furthermore, LC suppressed the expression of apoptosis‑associated genes (caspase-3) and inflammation‑associated genes [interleukin (IL)1, IL6, IL8 and cyclooxygenase‑2]. Conversely, LC promoted proliferating cell nuclear antigen, cyclin‑dependent kinase (CDK)2 and CDK4 expression, which may increase proliferation of HLECs that were incubated with H2O2. In addition, epithelial‑mesenchymal transition occurred upon ROS accumulation, whereas the effects of H2O2 on AQP1 and vimentin expression were reversed upon LC supplementation. Notably, this study revealed that LC restored the oxidant/antioxidant balance and protected against cell damage through the mitogen‑activated protein kinase signaling pathway. In conclusion, LC may serve a protective role in curbing oxidative damage and therefore may be considered a potential therapeutic agent for the treatment of cataracts.
    DOI:  https://doi.org/10.3892/ijmm.2019.4283
  28. Nutr Cancer. 2019 Jul 27. 1-12
    Zhu S, Wei L, Lin G, Tong Y, Zhang J, Jiang X, He Q, Lu X, Zhu DD, Chen YQ.
      Kudingcha is implicated in alleviating metabolic disorders in traditional Chinese medicine. However, the role of Kudingcha, one of the Ligustrum robustum species, in metabolic regulations and its antitumor activity in triple-negative breast cancer (TNBC) remains to be determined. Two breast cancer cell lines and immunocompetent and immunodeficient mice were used to evaluate the therapeutic effects of Kudingcha treatment. The production of reactive oxygen species (ROS) and glucose uptake were examined by flow cytometry. Metabolic shift was examined by metabonomics and western blot analysis. In this study, we found that aqueous extract of Kudingcha dose dependently inhibited cell growth and induced apoptosis in vitro and in vivo. Moreover, Kudingcha supplementation significantly reduced cancer metastasis. Kudingcha significantly inhibited glycolysis and glutamine metabolism. In addition, we demonstrated that the antitumor effects of Kudingcha were dependent on ROS production, which was increased by β-oxidation and oxidative phosphorylation. These findings provide a novel potential benefit of Kudingcha from targeting the cancer metabolism.
    DOI:  https://doi.org/10.1080/01635581.2019.1645865
  29. Arch Med Sci. 2019 Jul;15(4): 1001-1009
    Zhang Q, Bao J, Yang J.
      Introduction: Liver cancer is one of the most common malignancies across the globe and one of the major causes of cancer-related mortality. With limited available treatment options, there is an urgent need to look for new available options. Genistein is an important plant flavonoid and has been shown to possess tremendous pharmacological potential. The objective of the present study was therefore to evaluate the anticancer effect of the genistein.Material and methods: The antiproliferative activity and IC50 of genistein were determined by MTT assay. Reactive oxygen species (ROS) and cycle distribution were investigated by flow cytometry. Apoptosis was detected by DAPI and annexin V/IP staining. Cell migration was investigated by wound healing assay. Protein expression was estimated by western blotting.
    Results: MTT assay revealed that genistein reduced the cell viability of HepG2 cancer cells in a dose-dependent manner. Genistein also reduced the colony forming potential of the HepG2 cell concentration dependently. The IC50 of genistein was found to be 25 μM. Genistein caused G2/M cell cycle arrest and G2/M cells increased from 4.2% in the control to 56.4% at 100 μM concentration. Genistein prompted generation of significant (p < 0.01) amounts of ROS, ultimately favouring cell death. Genistein also triggered apoptosis which was associated with upregulation of cytosolic cytochrome c, Bax, cleaved caspase 3 and 9 expression and downregulation of Bcl-2 expression in HepG2 cells.
    Conclusions: We propose that genistein exhibits significant anticancer activity against liver cancer and therefore may prove beneficial in the management of liver cancer.
    Keywords:  HepG2; anticancer; genistein; liver cancer
    DOI:  https://doi.org/10.5114/aoms.2018.78742
  30. Zhongguo Zhong Yao Za Zhi. 2019 Jul;44(13): 2686-2690
    Wang JK, Wang LC, Jiang Y, Tu PF, Zeng KW.
      To investigate the inhibitory effects and mechanism of Cistanche tubulosa ethanol extract( CTEE) against oxygen-glucose deprivation/reperfusion( OGD/R)-induced PC12 cells neuronal injury. In this study,OGD/R-induced PC12 cells were used to explore the neuroprotective effects of CTEE( 12. 5,25,50 mg·L-1) by detecting cell viability with MTT assay,apoptosis with AO/EB and Hoechst 33258,mitochondrial membrane potential changes with JC-1 staining,mitochondrial oxidative stress with MitoSOX staining,as well as the apoptosis-related protein expression( PARP,cleaved PARP,caspase-3,cleaved caspase-3,Bax,Bcl-2) with Western blot. RESULTS:: showed that CTEE effectively protected OGD/R-induced neuronal injury and increased the survival rate of PC12 cells.AO/EB and Hoechst 33258 staining showed that CTEE could effectively inhibit apoptosis. Moreover,JC-1 and MitoSOX staining results showed that CTEE decreased mitochondrial stress and mitochondrial membrane potential imbalance in PC12 cells in a concentration-dependent manner. Meanwhile,CTEE could obviously suppress the activation of key proteins in mitochondrial apoptosis pathway such as caspase-3 and PARP,and significantly inhibit the rise of Bax and down-regulation of Bcl-2. In conclusion,CTEE has obvious protective effects on OGD/R-induced PC12 cells neuronal injury,potentially via inhibiting mitochondrial oxidative stress and apoptosis-related signaling pathway.
    Keywords:  Cistanche tubulosa; ethanol extract; mitochondri al pathway; neuronal injury; oxygen-glucose deprivation/reperfusion(OGD/R)
    DOI:  https://doi.org/10.19540/j.cnki.cjcmm.20181214.001
  31. Cell Stress Chaperones. 2019 Jul 30.
    Nash S, Johnstone J, Rahman MS.
      Global climate change is predicted to intensify thermal stress in marine and coastal organisms, affecting their development, growth, and reproductive functions. In this study, we performed histological observations on ovarian development, immunohistochemical analyses of ovarian heat shock protein-70 (HSP70), nitrotyrosine protein (NTP, an indicator of reactive nitrogen species (RNS)), and dinitrophenyl protein (DNP, an indicator of protein oxidation) expressions, in situ TUNEL assay for cellular apoptosis, biochemical analyses of ovarian caspase-3/7 activity and protein carbonyl (PC, a measure of reactive oxygen species (ROS)) contents, nitrate/nitrite (NOx) levels, and extrapallial fluid (EPF, an important body fluid) pH in the American oyster, Crassostrea virginica. Oysters were exposed to medium (28 °C) and high (32 °C) temperatures under controlled laboratory conditions for 1 week. Oysters exposed to higher temperatures significantly decreased the number and diameter of eggs, and EPF protein concentrations compared with controls (24 °C). In contrast, EPF pH, ovarian HSP70 mRNA levels, and protein expression were increased after heat exposure, consistent with increased ovarian apoptosis. The enhanced apoptosis in ovaries was associated with increased ovarian caspase-3/7 activity, PC contents, NOx levels, and NTP and DNP expressions in heat-exposed oysters. Collectively, these results suggest that higher temperatures drastically increase RNS and ROS levels, increasing incidence of apoptosis and subsequently reducing ovarian functions in oysters.
    Keywords:  Apoptosis; Global warming; Heat shock protein; Ovary; Oxidative stress; Oyster
    DOI:  https://doi.org/10.1007/s12192-019-01023-w
  32. Biomed Res Int. 2019 ;2019 1528278
    Li J, Wan W, Chen T, Tong S, Jiang X, Liu W.
      Oxidative stress and cardiomyocytes apoptosis were closely involved in the pathological process of doxorubicin- (Dox-) induced cardiac injury. MicroRNA-451 (miR-451) was mainly expressed in cardiomyocytes. However, the role of miR-451 in Dox-induced cardiac injury remained unclear. Our study aimed to investigate the effect of miR-451 on Dox-induced cardiotoxicity in mice. We established a Dox-induced cardiotoxicity model in the mice and manipulated miR-451 expression in the heart using a miR-451 inhibitor, which was injected every other day beginning at one day before Dox injection. Oxidative stress and apoptosis in the hearts were evaluated. miR-451 levels were significantly increased in Dox-treated mice or cardiomyocytes. miR-451 inhibition attenuated Dox-induced whole-body wasting and heart atrophy, reduced cardiac injury, restored cardiac function, and improved cardiomyocyte contractile function. Moreover, miR-451 inhibition reduced oxidative stress and cardiomyocytes apoptosis in vivo and in vitro. miR-451 inhibition increased the expression of calcium binding protein 39 (Cab39) and activated adenosine monophosphate activated protein kinase (AMPK) signaling pathway. A specific inhibitor of AMPK abolished the protection provided by miR-451 inhibition against cell injury in vitro. In conclusion, miR-451 inhibition protected against Dox-induced cardiotoxicity via activation of AMPK signaling pathway.
    DOI:  https://doi.org/10.1155/2019/1528278
  33. J Antibiot (Tokyo). 2019 Aug 02.
    Yokosawa T, Yamada M, Noguchi T, Suzuki S, Hirata Y, Matsuzawa A.
      Polymyxin B (PMB), a last-line antibiotic used against antibiotic-resistant superbugs, causes undesirable cytotoxic side effects. However, its mechanisms remain unknown. In this study, we unexpectedly found that caspase-3, a main executor of apoptosis, plays a protective role in PMB-induced cytotoxicity. Caspase-3 knockout (KO) cells exhibited higher susceptibility to PMB-induced cytotoxicity compared with wild-type (WT) cells, accompanied by increased levels of reactive oxygen species (ROS). Interestingly, co-treatment with the antioxidant N-acetylcysteine (NAC) rescued cell viability to a similar extent as WT cells. Furthermore, PMB failed to facilitate the processing of inactive caspase-3 (pro-caspase-3) into active forms, suggesting that pro-caspase-3 nonenzymatically suppresses PMB-driven ROS accumulation and its cytotoxicity. Thus, our findings that demonstrate the potential ability of PMB to stimulate ROS generation, but which is normally masked by pro-caspase-3-dependent mechanisms, may provide novel insights into the mechanisms of PMB-induced side effects.
    DOI:  https://doi.org/10.1038/s41429-019-0216-6
  34. Oxid Med Cell Longev. 2019 ;2019 2606120
    Zalewska A, Ziembicka D, Żendzian-Piotrowska M, Maciejczyk M.
      Oxidative stress plays a crucial role in the salivary gland dysfunction in insulin resistance; however, the cause of increased free radical formation in these conditions is still unknown. Therefore, the aim of the study was to investigate the effect of high-fat diet (HFD) on the mitochondrial respiratory system, prooxidant enzymes, ROS production, and nitrosative/oxidative stress in the submandibular and parotid glands of rats. The experiment was performed on male Wistar rats divided into two groups (n = 10): control and HFD. The 8-week feeding of HFD affects glucose metabolism observed as significant increase in plasma glucose and insulin as well as HOMA-IR as compared to the control rats. The activity of mitochondrial Complex I and Complex II+III was significantly decreased in the parotid and submandibular glands of HFD rats. Mitochondrial cytochrome c oxidase (COX) activity and the hydrogen peroxide level were significantly increased in the parotid and submandibular glands of the HFD group as compared to those of the controls. HFD rats also showed significantly lower reduced glutathione (GSH) and reduced : oxidized glutathione (GSH : GSSG) ratio, as well as a higher GSSG level in the parotid glands of HFD rats. The activity of NADPH oxidase, xanthine oxidase, and levels of oxidative/nitrosative stress (malonaldehyde, nitric oxide, nitrotyrosine, and peroxynitrite) and inflammation/apoptosis (interleukin-1β and caspase-3) biomarkers were statistically elevated in the HFD group in comparison to the controls. HFD impairs mitochondrial function in both types of salivary glands by enhancing ROS production, as well as stimulating inflammation and apoptosis. However, free radical production, protein nitration, and lipid peroxidation were more pronounced in the parotid glands of HFD rats.
    DOI:  https://doi.org/10.1155/2019/2606120
  35. Nucl Med Commun. 2019 Jul 29.
    Akbulut A, Bilge Keseroglu B, Koca G, Nedim Yuceturk C, Cem Ozgur B, Surer H, Ogus E, Yumuşak N, Karakaya J, Korkmaz M.
      PURPOSE: Renal ischemia-reperfusion injury (RIRI) may occur secondary to several reasons leading to renal failure. Coenzyme-Q10 (CoQ10) is a well-known antioxidant. However, the effects CoQ10 against RIRI have not been evaluated. Our aim was to evaluate protective effects of CoQ10 to renal ischemia-reperfusion by biochemical, immunohistochemical and scintigraphic findings.METHODS: Thirty Wistar-albino rats were randomly separated into groups of 10; Group Sham; Group ischemia-reperfusion (IR) had left renal pedicle clamping; Group CoQ10+IR had IR and CoQ10. Twenty-four hours later after reperfusion, scintigraphy was performed and after that, rats were sacrificed. To demonstrate effects of RIRI, serum urea and creatinine levels and tissue levels oxidative stress markers were evaluated. Both kidneys were subjected to histopathological evaluation and to confirm RIRI-induced immunohistochemical aspects of apoptosis, terminal-deoxynucleotidyl-transferase mediated-deoxyuridine-triphosphate-nick-end-labeling assay and caspase-3 were assessed.
    RESULTS: Tissue oxidative stress, histopathologic changes, apoptosis scores and quantitative scintigraphic parameters were significantly higher in Group IR compared with Group Sham. Although tissue oxidative stress levels and histopathologic changes were not significant, quantitative scintigraphic parameters of contralateral kidney of Group IR were significantly increased. Compared with Group IR, Group CoQ10+IR presented decreased tissue oxidative stress levels; decreased scores of histopathology and apoptosis; and decreased quantitative scintigraphic parameters with increased split renal function in ischemic kidney.
    CONCLUSIONS: Our results suggest that other than its antioxidant properties, CoQ10 shows antiperoxidative, antiapoptotic and antiinflammatory potential in protecting renal functioning of ischemic kidney. Furthermore, our results show that renal scintigraphy is a feasible method to detect early changes in renal functioning after RIRI.
    DOI:  https://doi.org/10.1097/MNM.0000000000001070
  36. J Am Heart Assoc. 2019 Aug 06. 8(15): e012047
    Nishi M, Ogata T, Cannistraci CV, Ciucci S, Nakanishi N, Higuchi Y, Sakamoto A, Tsuji Y, Mizushima K, Matoba S.
      Background Ischemia/reperfusion (I/R) injury is a critical issue in the development of treatment strategies for ischemic heart disease. MURC (muscle-restricted coiled-coil protein)/Cavin-4 (caveolae-associated protein 4), which is a component of caveolae, is involved in the pathophysiology of dilated cardiomyopathy and cardiac hypertrophy. However, the role of MURC in cardiac I/R injury remains unknown. Methods and Results The systems network genomic analysis based on PC-corr network inference on microarray data between wild-type and MURC knockout mouse hearts predicted a network of discriminating genes associated with reactive oxygen species. To demonstrate the prediction, we analyzed I/R-injured mouse hearts. MURC deletion decreased infarct size and preserved heart contraction with reactive oxygen species-related molecule EGR1 (early growth response protein 1) and DDIT4 (DNA-damage-inducible transcript 4) suppression in I/R-injured hearts. Because PC-corr network inference integrated with a protein-protein interaction network prediction also showed that MURC is involved in the apoptotic pathway, we confirmed the upregulation of STAT3 (signal transducer and activator of transcription 3) and BCL2 (B-cell lymphoma 2) and the inactivation of caspase 3 in I/R-injured hearts of MURC knockout mice compared with those of wild-type mice. STAT3 inhibitor canceled the cardioprotective effect of MURC deletion in I/R-injured hearts. In cardiomyocytes exposed to hydrogen peroxide, MURC overexpression promoted apoptosis and MURC knockdown inhibited apoptosis. STAT3 inhibitor canceled the antiapoptotic effect of MURC knockdown in cardiomyocytes. Conclusions Our findings, obtained by prediction from systems network genomic analysis followed by experimental validation, suggested that MURC modulates cardiac I/R injury through the regulation of reactive oxygen species-induced cell death and STAT3-meditated antiapoptosis. Functional inhibition of MURC may be effective in reducing cardiac I/R injury.
    Keywords:  apoptosis; caveolae; ischemia reperfusion injury; reactive oxygen species; systems biology
    DOI:  https://doi.org/10.1161/JAHA.119.012047
  37. Zhen Ci Yan Jiu. 2019 Jun 25. 44(6): 412-8
    Sun Y, Tu Y, Guo Y, Jiang HL, Li YH, Wang Y, Zhao BC, Gao XZ, Li X, Feng SX, Li YF.
      OBJECTIVE: To investigate the effect of acupuncture on behavior changes and the expression of reactive oxygen species (ROS), cytochrome C, cysteine-containing aspartate-specific proteases-3 (caspase-3), apoptosis inducing factor (AIF) in the hippocampus of rats exposed to chronic restraint stress (CRS), so as to provide experimental evidence for the prevention and treatment of depression.METHODS: Thirty two male SD rats were equally randomized into control, model, acupuncture and medication (Fluoxetine) groups according to the random number table. The depression model was established by chronic psychological stress stimulation (fasting, water deprivation, chronic restraint combined with solitary raising for 28 days). Manual acupuncture stimulation (2 circles/s) was applied to "Baihui" (GV20), "Yintang" (GV29) and bilateral "Sanyinjiao" (SP6) for 2 min, followed by retaining the needles for 20 min. Rats in the medication group were treated by gavage of Fluoxetine (0.18 mg/mL, 1 mL/100 g body weight). The treatment was conducted once daily for 28 days. The depression-like behavior was evaluated by using open-field test, sucrose preference test, and body weight. The content of ROS of the right hippocampus tissue was detected by using dichloro-dihydro-fluorescein diacetate (DCFH-DA) assay, and the expression levels of cytochrome C, caspase-3 and AIF proteins of hippocampus were determined by using Western blot.
    RESULTS: After 28 days' stress procedure, the depression rats in the model group showed a significant decrease in the numbers of crossing and rearing of open-field test, sucrose preference index and the body weight compared to the control group (P<0.01). The content of ROS and expression levels of cytochrome C, caspase-3 and AIF in the hippocampus of the model group were significantly increased relevant to the control group (all P<0.01). After the intervention and compared with the model group, the decreased levels of the numbers of crossing and rearing, body weight and sucrose preference index, and the up-regulated levels of content of ROS and expression of cytochrome C, caspase-3 and AIF proteins were all reversed in both acupuncture and medication groups (P<0.01,P<0.05), displaying an anti-depressant effect. The effect of acupuncture was significantly superior to that of medication in up-regulating the numbers of crossing and rearing (P<0.01, P<0.05), and in down-regulating the expression of caspase-3 (P<0.01). No significant differences were found between the medication and acupuncture groups in increasing the body weight and sucrose preference index, and in down-regulating the expression of cytochrome C and AIF proteins (P>0.05).
    CONCLUSION: Acupuncture intervention can significantly reduce ROS content and expression level of cytochrome C, caspase-3 and AIF proteins in the hippocampus possibly by inhibiting the chronic psychological stress-hippocampal oxidative stress-mitochondrial apoptotic pathway in psychological stress-induced depression rats, which may contribute to its effect in relieving depression.
    Keywords:  Acupuncture; Cell apoptosis; Chronic psychological stress; Depression; Hippocampus; Oxidative stress
    DOI:  https://doi.org/10.13702/j.1000-0607.190098
  38. J Food Biochem. 2019 Jul;43(7): e12836
    Cao L, Zhi D, Han J, Kumar Sah S, Xie Y.
      Gentamicin (GM) is an antibiotic related to aminoglycoside group that is used in treating Gram-negative bacterial infections. However, treatment with gentamicin is considered to be limited as it induces an oxidative stress-mediated apoptosis in kidney which causes a nephrotoxicity. Metformin is a well-known biguanide that is used for treating diabetes mellitus, especially type 2. Supplement with plant metabolites or natural antioxidants produce a protective activity against many types of diseases in vivo. Curcumin is a main medicinal constituent of Curcuma longa, has reported for number of biological effects, such as antioxidant, anti-inflammatory, and antitumor. The study aims at evaluating the metformin and curcumin alone or in combination on nephrotoxicity induced by GM. The outcome of the study shows that both metformin and curcumin, when used unaided, were effectively decreasing GM-induced nephrotoxicity. The two drugs combination was showed synergistic effect in ameliorating a GM-induced kidney injury, as supported by expressively improved renal dysfunction. Metformin and curcumin showed strong protection against oxidative stress in GM treated animals through decreasing the activities and expression of various antioxidative enzymes. Moreover, combination of two drugs showed an anti-inflammatory response through reducing a level of pro-inflammatory cytokines including tumor necrosis factor-alpha, interleukin 1-beta, and interleukin 6 in GM intoxicated group of animals. Furthermore, GM agitated apoptosis was affectedly diminished by the combinational treatment of metformin and curcumin via down-regulating activity of cleaved Caspase-3 and pro-apoptotic factor Bax, whereas increasing anti-apoptotic factor Bcl-2 signaling pathways. The above results suggested that combinational treatment of metformin and curcumin might be have a synergizing effect and substantial potential against nephrotoxicity induced by GM. PRACTICAL APPLICATIONS: Curcumin and metformin combination exhibited substantial synergistic effect against GM-induced nephrotoxicity through reducing oxidative stress, inflammation, as well as apoptosis in kidney cells. Therefore, the method of combination of curcumin and metformin might be functional to treat or inhibit GM prompted nephrotoxicity in future.
    Keywords:  apoptosis; curcumin; gentamicin; metformin; nephrotoxicity; oxidative stress
    DOI:  https://doi.org/10.1111/jfbc.12836
  39. J Food Biochem. 2019 Jul 31. e12999
    Youn Y, Jeon SH, Jin HY, Che DN, Jang SI, Kim YS.
      Neurodegenerative diseases are major threats to human health. Here, through fluorescence, colorimetric, immunoblotting, spectroscopy, and laser scanning confocal microscopic techniques, we investigated the neuroprotective properties of chlorogenic acid-rich Solanum melongena extracts (SM extract) in rotenone-induced PC-12 cell death. The results showed that rotenone caused apoptosis to PC-12 cells by elevating Bax/Bcl-2 ratio and increasing caspase-3 activity. Rotenone also increased ROS in cells while suppressing SOD and catalase activities. This resulted in the depletion of ATP in cells by blocking mitochondria complex I activity. Pretreatment of the cells with SM extract at concentrations of 100, 250, and 500 μg/ml before incubation for 24 hr with rotenone significantly prevented apoptosis, decreased ROS, and increased ATP production in the cells. SM extract upregulated SOD and catalase activities in the cells. These results unveil evidence that SM extract content neuroprotective properties that can be exploited to prevent and treat neurodegenerative diseases. PRACTICAL APPLICATIONS: Solanum melongena eggplant is a popular ingredient in many traditional recipes and is well known in Asia for its medicinal benefits. Despite numerous scientific reports of the potential health benefits of this plant, reports on its effects in neurodegenerative diseases is still lacking. This pilot study demonstrates that S. melongena eggplant can protect against neurotoxicity in neurodegenerative diseases. The results of this research serves as a base for further research on eggplant that will result in its usage on a larger scale as functional food materials.
    Keywords:   Solanum melongena ; PC-12 cell; apoptosis; chlorogenic acid; neuroprotection; oxidative stress; rotenone
    DOI:  https://doi.org/10.1111/jfbc.12999
  40. J Vet Med Sci. 2019 Aug 01.
    Shah MA, Park DJ, Kang JB, Kim MO, Koh PO.
      Baicalin is a plant-derived flavonoid that has anti-inflammatory and anti-oxidative effects. We investigated an anti-inflammatory effect of baicalin against lipopolysaccharide (LPS)-induced damage in cerebral cortex. Adult mice were divided into control, LPS-treated, and LPS and baicalin co-treated animals. LPS (250 µg/kg/day) and baicalin (10 mg/kg/day) were intraperitoneally injected for 7 days. LPS treatment induced histopathological changes in cerebral cortex, whereas baicalin protected neuronal cells against LPS toxicity. Moreover, baicalin treatment attenuated LPS-induced increases of reactive oxygen species and oxidative stress in cerebral cortices. Ionized calcium binding adaptor molecule-1 (Iba-1) and glial fibrillary acidic protein (GFAP) are known as markers of activated microglia and astrocyte, respectively. Results of Western blot and immunofluorescence staining showed that LPS exposure induces increases of Iba-1 and GFAP expressions, whereas baicalin alleviates LPS-induced increases of these proteins. Baicalin also prevented LPS-induced increase of nuclear factor kappa B (NF-κB). LPS treatment led to increases of pro-inflammatory factors including, interleukin-1β (IL-1β) and tumor necrosis factor-α (TNF-α). Increases of these pro-inflammatory mediators were attenuated in baicalin co-treated animals. These results demonstrated that baicalin regulates neuroglia activation and modulates inflammatory factors in LPS-induced neuronal injury. Thus, our findings suggest that baicalin exerts a neuroinflammatory effect against LPS-induced toxicity through decreasing oxidative stress and inhibiting NF-κB mediated inflammatory factors, such as IL-1β and TNF-α.
    Keywords:  baicalin; lipopolysaccharide; neuroinflammation; oxidative stress; reactive oxygen species
    DOI:  https://doi.org/10.1292/jvms.19-0281
  41. IUBMB Life. 2019 Jul 30.
    Davargaon RS, Sambe AD, Muthangi V V S.
      Redox imbalance due to hyperglycemia is a causative factor for an increased generation of reactive oxygen species (ROS) that leads to mitochondrial dysfunction and the release of cytochrome-c. The aim of the present study is to elucidate the functional role of oxidative stress (OS) in the induction of apoptosis in H9c2 cells in the hyperglycemic state through glucose transporter-4 (GLUT-4) regulation and antioxidant status. H9c2 cells were incubated with 15, 24, and 33 mM glucose for 24, 48, and 72 hr to induce hyperglycemic stress. Hyperglycemic episodes have significantly influenced GLUT-4 mRNA regulation, depleted glutathione (GSH) and its associated enzymes, reduced cellular antioxidant enzymes (AOEs), caused nuclear condensation, and induced apoptosis by activating caspase-9 and 3 and annexin V binding in a concentration and duration-dependent manner. Trolox pretreatment significantly enhanced the GLUT-4 mRNA and antioxidant defense mechanism, suppressed nuclear condensation, and prevented cytochrome-c release, thereby reducing mitochondrial-dependent apoptosis. The present study shows that the toxic effect of high glucose is significantly regulated and that OS induction can be prevented through a water-soluble vitamin E analog "Trolox" treatment.
    Keywords:  GLUT-4; GSH; apoptosis; hyperglycemia; oxidative stress; rat myocardial cells
    DOI:  https://doi.org/10.1002/iub.2133
  42. Cancer Med. 2019 Jul 31.
    Chikamatsu S, Saijo K, Imai H, Narita K, Kawamura Y, Katoh T, Ishioka C.
      Liphagal, isolated from the marine sponge Aka coralliphaga, exhibits phosphatidylinositol 3-kinase alpha (PI3Kα) inhibitory activity and cytotoxic effects in human cancer cells. Siphonodictyal B, the biogenetic precursor of liphagal, also has PI3K inhibitory activity. However, its cytotoxic or antitumor activities have not been evaluated. In this study, we demonstrated that siphonodictyal B inhibits several kinases such as CDK4/6, CDK7, and PIM2 in addition to PI3K in vitro and that siphonodictyal B exhibits more potent cytotoxic effects than liphagal against human colon cancer cell lines. Furthermore, treatment with siphonodictyal B resulted in increased PARP cleavage, a larger sub-G1 fraction, and a larger annexin V-positive cell population, all of which are indicative of apoptosis induction. As a mechanism of apoptosis induction, we found that siphonodictyal B activates the p38 MAPK pathway, leading the upregulation of proapoptotic factors. Moreover, siphonodictyal B increased ROS levels, thus promoting p38 MAPK pathway activation. NAC, an ROS scavenger, almost completely reversed both the cytotoxic and p38 MAPK pathway-activating effects of siphonodictyal B. These results indicate that the p38 MAPK pathway might be involved downstream of ROS signaling as part of the mechanism of siphonodictyal B-induced apoptosis. Finally, siphonodictyal B displayed antitumor effects in a human colon cancer xenograft mouse model and increased p38 phosphorylation in tumor tissue. These results suggest that siphonodictyal B could serve as the basis of a novel anticancer drug.
    Keywords:  liphagal; p38 MAPK pathway; reactive oxygen species; siphonodictyal B; terpenoid
    DOI:  https://doi.org/10.1002/cam4.2409
  43. Biotech Histochem. 2019 Aug 02. 1-14
    Yeşil H, Tuğlu I.
      Heart attack and oxygen deficiency may cause necrosis in the brain and other tissues. We investigated the histopathological effects of nitric oxide (NO) on ischemia/reperfusion in lung and hippocampus using a rat brain bilateral occlusion ischemia model. Male rats were assigned to sham (SH), ischemic preconditioning (PC), global ischemia (GI) and ischemic reperfusion (IR) groups. Before ischemia was induced, blood was drawn to induce hypovolemic hypotension and for blood gas testing. After sacrifice, samples of hippocampus were harvested. Sections were examined using hematoxylin and eosin (H & E) staining and immunostaining using primary antibodies for GFAP, S100β, iNOS, eNOS and the TUNEL method. Following ischemia, we found evidence of gliosis induced oxidative stress and apoptosis in the hippocampus. No significant differences were detected between the SH and PC groups. In the GI and IR groups, apoptosis and necrosis were observed in the hippocampus. Lung sections were stained with H & E and Masson's trichrome (MT) and immunostained for iNOS and eNOS. The TUNEL method was used to detect apoptosis. Interstitial edema, vascular congestion, intra-alveolar hemorrhage, perivascular edema, neutrophil infiltration and disruption of alveoli were observed after global ischemia and ischemic reperfusion. Inflammatory cells were detected in the connective tissue. The IR and GI groups exhibited significantly more apoptotic cells than the SH or PC groups. Free radicals, such as nitric oxide (NO), that appear following ischemia and reperfusion in the brain may also injure the lungs. Increased NO in both lung and brain tissue suggests that apoptosis in these organs can be induced by reactive nitrogen species.
    Keywords:  Apoptosis; brain; histology; immunohistochemistry; ischemia; lung; oxidative stress; rats; reperfusion
    DOI:  https://doi.org/10.1080/10520295.2019.1601768
  44. BMC Ophthalmol. 2019 Aug 02. 19(1): 168
    Chen JL, Hung CT, Keller JJ, Lin HC, Wu YJ.
      BACKGROUND: Age-related macular degeneration (AMD) is the primary cause of blindness and severe vision loss in developed countries and is responsible for 8.7% of blindness globally. Ultraviolet radiation can induce DNA breakdown, produce reactive oxygen species, and has been implicated as a risk factor for AMD. This study investigated the effects of UVA radiation on Human retinal pigment epithelial cell (ARPE-19) growth and protein expression.METHODS: ARPE-19 cells were irradiated with a UVA lamp at different doses (5, 10, 20, 30 and 40 J/cm2) from 10 cm. Cell viability was determined by MTT assay. Visual inspection was first achieved with inverted light microscopy and then the DeadEnd™ Fluorometric TUNEL System was used to observe nuclear DNA fragmentation. Flow cytometry based-Annexin V-FITC/PI double-staining was used to further quantify cellular viability. Mitochondrial membrane potential was assessed with JC-1 staining. 2D electrophoresis maps of exposed cells were compared to nonexposed cells and gel images analyzed with PDQuest 2-D Analysis Software. Spots with greater than a 1.5-fold difference were selected for LC-MS/MS analysis and some confirmed by western blot. We further investigated whether caspase activation, apoptotic-related mitochondrial proteins, and regulators of ER stress sensors were involved in UVA-induced apoptosis.
    RESULTS: We detected 29 differentially expressed proteins (9 up-regulated and 20 down-regulated) in the exposed cells. Some of these proteins such as CALR, GRP78, NPM, Hsp27, PDI, ATP synthase subunit alpha, PRDX1, and GAPDH are associated with anti-proliferation, induction of apoptosis, and oxidative-stress protection. We also detected altered protein expression levels among caspases (caspase 3 and 9) and in the mitochondrial (cytosolic cytochrome C, AIF, Mcl-1, Bcl-2, Bcl-xl, Bax, Bad, and p-Bad) and ER stress-related (p-PERK, p-eIF2α, ATF4 and CHOP) apoptotic pathways.
    CONCLUSIONS: UVA irradiation suppressed the proliferation of ARPE-19 cells in a dose-dependent manner, caused quantitative loses in transmembrane potential (ΔΨm), and induced both early and late apoptosis.
    Keywords:  Apoptosis; Mitochondrial dysfunction; Proteomic; Retinal pigment epithelium cells; UVA
    DOI:  https://doi.org/10.1186/s12886-019-1151-9
  45. Free Radic Biol Med. 2019 Jul 26. pii: S0891-5849(18)32514-0. [Epub ahead of print]143 25-46
    Nyunt T, Britton M, Wanichthanarak K, Budamagunta M, Voss JC, Wilson DW, Rutledge JC, Aung HH.
      Elevation of blood triglycerides, primarily triglyceride-rich lipoproteins (TGRL), is an independent risk factor for cardiovascular disease and vascular dementia (VaD). Accumulating evidence indicates that both atherosclerosis and VaD are linked to vascular inflammation. However, the role of TGRL in vascular inflammation, which increases risk for VaD, remains largely unknown and its underlying mechanisms are still unclear. We strived to determine the effects of postprandial TGRL exposure on brain microvascular endothelial cells, the potential risk factor of vascular inflammation, resulting in VaD. We showed in Aung et al., J Lipid Res., 2016 that postprandial TGRL lipolysis products (TL) activate mitochondrial reactive oxygen species (ROS) and increase the expression of the stress-responsive protein, activating transcription factor 3 (ATF3), which injures human brain microvascular endothelial cells (HBMECs) in vitro. In this study, we deployed high-throughput sequencing (HTS)-based RNA sequencing methods and mito stress and glycolytic rate assays with an Agilent Seahorse XF analyzer and profiled the differential expression of transcripts, constructed signaling pathways, and measured mitochondrial respiration, ATP production, proton leak, and glycolysis of HBMECs treated with TL. Conclusions: TL potentiate ROS by mitochondria which activate mitochondrial oxidative stress, decrease ATP production, increase mitochondrial proton leak and glycolysis rate, and mitochondria DNA damage. Additionally, CPT1A1 siRNA knockdown suppresses oxidative stress and prevents mitochondrial dysfunction and vascular inflammation in TL treated HBMECs. TL activates ATF3-MAPKinase, TNF, and NRF2 signaling pathways. Furthermore, the NRF2 signaling pathway which is upstream of the ATF3-MAPKinase signaling pathway, is also regulated by the mitochondrial oxidative stress. We are the first to report differential inflammatory characteristics of transcript variants 4 (ATF3-T4) and 5 (ATF3-T5) of the stress responsive gene ATF3 in HBMECs induced by postprandial TL. Specifically, our data indicates that ATF3-T4 predominantly regulates the TL-induced brain microvascular inflammation and TNF signaling. Both siRNAs of ATF3-T4 and ATF3-T5 suppress cells apoptosis and lipotoxic brain microvascular endothelial cells. These novel signaling pathways triggered by oxidative stress-responsive transcript variants, ATF3-T4 and ATF3-T5, in the brain microvascular inflammation induced by TGRL lipolysis products may contribute to pathophysiological processes of vascular dementia.
    Keywords:  Activating transcription factor 3; Brain microvascular endothelial cells; Inflammation; Lipolysis; Mitochondrial oxidative stress; RNA-Seq; Triglyceride-rich lipoproteins
    DOI:  https://doi.org/10.1016/j.freeradbiomed.2019.07.024
  46. J Inorg Biochem. 2019 Jul 19. pii: S0162-0134(19)30333-2. [Epub ahead of print]199 110789
    Almeida J, Roma-Rodrigues C, Mahmoud AG, Guedes da Silva MFC, Pombeiro AJL, Martins LMDRS, Baptista PV, Fernandes AR.
      The water-soluble 1D helical coordination polymer [Ag(Tpms)]n (1) [Tpms = tris(pyrazolyl)methane sulfonate, -O3SC(pz)3; pz = pyrazolyl] was synthesized and fully characterized, its single-crystal X-ray diffraction analysis revealing the ligand acting as a bridging chelate N3-donor ligand. The antiproliferative potential of 1 was performed on two human tumour cell lines, A2780 and HCT116, and in normal fibroblasts, with a much higher effect in the former cell line (IC50 of 0.04 μM) as compared to the latter cell line and to normal fibroblasts. Compound 1 does not alter cell cycle progression but interferes with the adherence of A2780 cells triggering cell apoptosis. Apoptosis appears to occur via the extrinsic pathway (no changes in mitochondria membrane potential, reactive oxygen species (ROS) and pro-apoptotic (B-cell lymphoma 2 (BCL-2) associated protein (BAX))/anti-apoptotic (BCL-2) ratio) being this hypothesis also supported by the presence of silver mainly in the supernatants of A2780 cells. Results also indicated that cell death via autophagy was triggered. Proteomic analysis allowed us to confirm that compound 1 is able to induce a stress response in A2780 cells that is related with its antiproliferative activity and the trigger of apoptosis.
    Keywords:  2-D electrophoresis; Antiproliferative activity; Cell death; Ovarian carcinoma; Silver complexes
    DOI:  https://doi.org/10.1016/j.jinorgbio.2019.110789
  47. Epigenomics. 2019 Jul 29.
    Shen L, Zhao K, Li H, Ning B, Wang W, Liu R, Zhang Y, Zhang A.
      Aim: To investigate the effect of UBE2T gene on radiotherapy for osteosarcoma. Materials & methods: Gene Expression Omnibus database, RT-qPCR and immunohistochemical analysis were performed. Cell proliferation and cell cycle experiments were conducted after knockdown of UBE2T. Cell scratch, reactive oxygen species production and apoptosis experiments were conducted after the combination of radiotherapy and UBE2T silencing. Then the xenograft mode was further conducted. Results: UBE2T was highly expressed in human osteosarcoma. Suppression of UBE2T inhibited osteosarcoma cell proliferation and induced cell cycle arrest at the G2/M phase. Downregulation of UBE2T combined with radiation can substantially inhibit clonal formation and migration, and promote apoptosis of osteosarcoma cells in vitro and in vivo. Conclusion: UBE2T downregulation can enhance the radiosensitivity of osteosarcoma in vitro and in vivo.
    Keywords:  ; GEO database; cell apoptosis; cell cycle arrest; cell migration; cell proliferation; human osteosarcoma; radiosensitization
    DOI:  https://doi.org/10.2217/epi-2019-0125
  48. Arch Med Sci. 2019 Jul;15(4): 1104-1112
    Eken MK, Ersoy GS, Kaygusuz EI, Devranoğlu B, Takır M, Çilingir ÖT, Çevik Ö.
      Introduction: Etanercept has been widely used in autoimmune diseases for blocking tumor necrosis factor α (TNF-α), which is an inflammatory cytokine. The anti-apoptotic and anti-inflammatory effects of etanercept against ischemia/reperfusion (I/R) injury have been shown for several tissues in rat studies, but to the best of our knowledge, there are no reports on its protective effects following similar injury in ovarian tissue. The aim of this study was to investigate whether etanercept has beneficial effects on ovarian I/R injury, as well as on ovarian reserve.Material and methods: Twenty-four rats were randomly divided into four groups (n = 6/group): sham (laparotomy only); sham + etanercept; I/R; and I/R + etanercept. Ischemia was induced for 3 h by twisting the ovary, and 24 h after detorsion the ovarian tissues were collected to evaluate histopathologic changes, glutathione (GSH), malondialdehyde (MDA), myeloperoxidase (MPO), and superoxide dismutase (SOD) concentrations for oxidative stress, 8-hydroxy-2'-deoxyguanosine (8-OHdG) for DNA damage, caspase-3 activity for apoptosis and ovarian follicle counts. To measure anti-Mullerian hormone (AMH), serum samples were drawn before and after surgery.
    Results: Tissue GSH and SOD levels were significantly higher, while MDA and MPO levels were significantly lower in the I/R + etanercept group than in the I/R group (p < 0.05, p < 0.01, respectively). Tissue 8-OHdG and caspase-3 activity were significantly lower in the I/R+etanercept group than in the I/R group (p < 0.05, p < 0.01, respectively). Preoperative and postoperative AMH levels were compared and there was a significant reduction in the I/R and I/R + etanercept groups (p < 0.001, p < 0.001). The reduction of AMH in the I/R + etanercept group was significantly lower than in the I/R group. The primordial, preantral and small antral follicle numbers were also significantly higher in the I/R + etanercept group compared to the I/R group (p < 0.001, p < 0.001, p < 0.005, respectively).
    Conclusions: Etanercept attenuated inflammation and related oxidative stress and also helped to preserve ovarian reserve following ovarian I/R damage.
    Keywords:  apoptosis; etanercept; inflammation; ischemia/reperfusion injury; ovary; oxidative injury
    DOI:  https://doi.org/10.5114/aoms.2017.72406
  49. J Surg Res. 2019 Jul 24. pii: S0022-4804(19)30470-6. [Epub ahead of print]244 547-557
    Mao Y, Chen X, Xia Y, Xie X.
      BACKGROUND: Ischemia-reperfusion (IR) injury is a main cause to and the mechanism of necrosis after flap transplantation. Researches were hardly conducted on the role and possible mechanism of keratinocyte growth factor (KGF) in association with IR flap injury.MATERIALS AND METHODS: A CoCl2-stimulated hypoxia cell model was established to investigate the effects of KGF on cell viability, apoptosis, cell cycle, and reactive oxygen species level. The experiments were performed by cell counting kit-8 and flow cytometry as required. Meanwhile, the expressions of cell cycle-related and nuclear factor E2-related factor 2 (Nrf2) signaling-related genes were determined using quantitative real-time PCR and Western blot. The right dorsolateral areas of Institute of Cancer Research mice were marked as flaps, the pedicle of which formed an IR process through clamping and loosening. Tissue morphologies were observed using hematoxylin and eosin staining 24 h after the surgery. The effects of KGF on cell apoptosis and associated genes expressions were studied by terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling, immunohistochemistry, and Western blot.
    RESULTS: HaCAT cells treated with 40 μM CoCl2 could not only reduce cell viability, promote cell apoptosis, arrest G1 phase of cell cycle and increase the activity of reactive oxygen species but also downregulate the expressions of c-myc, c-fos, transforming growth factor-α, Nrf2, heme oxygenase-1, and gamma-glutamyl cysteine synthetase. Additional recombinant human KGF, on one hand, could protect the cells from hypoxia injury. On the other hand, recombinant human KGF could significantly inhibit cell apoptosis, increase KGF activity, and increase the Nrf2, heme oxygenase-1, and gamma-glutamyl cysteine synthetase proteins levels in IR flap tissues.
    CONCLUSIONS: KGF played an important role in protecting mice flaps from IR injury, and the possible mechanism was involved in activating the Nrf2 signaling.
    Keywords:  Flap; Ischemia-reperfusion; Nrf2 signaling; Repair; rhKGF
    DOI:  https://doi.org/10.1016/j.jss.2019.06.078
  50. Int J Nanomedicine. 2019 ;14 4991-5015
    Daems N, Penninckx S, Nelissen I, Van Hoecke K, Cardinaels T, Baatout S, Michiels C, Lucas S, Aerts A.
      Purpose: This study evaluates the cytotoxicity of AuNPs coated with polyallylamine (AuNPs-PAA) and conjugated or not to the epidermal growth factor receptor (EGFR)-targeting antibody Cetuximab (AuNPs-PAA-Ctxb) in normal human kidney (HK-2), liver (THLE-2) and microvascular endothelial (TIME) cells, and compares it with two cancer cell lines that are EGFR-overexpressing (A431) or EGFR-negative (MDA-MB-453). Results: Conjugation of Cetuximab to AuNPs-PAA increased the AuNPs-PAA-Ctxb interactions with cells, but reduced their cytotoxicity. TIME cells exhibited the strongest reduction in viability after exposure to AuNPs-PAA(±Ctxb), followed by THLE-2, MDA-MB-453, HK-2 and A431 cells. This cell type-dependent sensitivity was strongly correlated to the inhibition of thioredoxin reductase (TrxR) and glutathione reductase (GR), and to the depolarization of the mitochondrial membrane potential. Both are suggested to initiate apoptosis, which was indeed detected in a concentration- and time-dependent manner. The role of oxidative stress in AuNPs-PAA(±Ctxb)-induced cytotoxicity was demonstrated by co-incubation of the cells with N-acetyl L-cysteine (NAC), which significantly decreased apoptosis and mitochondrial membrane depolarization. Conclusion: This study helps to identify the cells and tissues that could be sensitive to AuNPs and deepens the understanding of the risks associated with the use of AuNPs in vivo.
    Keywords:  Cetuximab; EGFR; cytotoxicity; oxidative stress
    DOI:  https://doi.org/10.2147/IJN.S203546
  51. Phytomedicine. 2019 Jul 20. pii: S0944-7113(19)30203-X. [Epub ahead of print]63 153037
    Ouyang XL, Qin F, Huang RZ, Liang D, Wang CG, Wang HS, Liao ZX.
      BACKGROUND: Non-Small-Cell Lung Cancer (NSCLC) is the most-frequent cause of cancer death, and novel chemotherapeutic drugs for treating NSCLC are urgently needed. 2α, 3α, 23-trihydroxy-13α, 27-cyclours-11-en-28-oic acid (euscaphic acid G) is a new hexacyclic triterpene acid isolated by our group from Glechoma longituba (Nakai) Kupr. However, the underlying mechanisms responsible for the anticancer effects of hexacyclic triterpene acid have not been elucidated.PURPOSE: In the present work, we evaluated growth inhibitory effect of the new isolated hexacyclic triterpene acid and explored the underlying molecular mechanisms.
    METHODS/STUDY DESIGNS: Herbs were extracted and constituents were purified by chromatographic separation, including silica gel, ODS, MCI, Sephadex LH-20 and preparative HPLC. The compound structures were elucidated by the use of UV, NMR and MS spectral data. The anticancer activity of euscaphic acid G was evaluated by MTT assay. Cell cycle, apoptosis, reactive oxygen species and mitochondrial membrane potential were determined by flow cytometry. To display the possible mechanism of euscaphic acid G on NCI-H460 cells, RT-PCR, immunofluorescence and Western blot analysis were carried out.
    RESULTS: A new hexacyclic triterpene acid, euscaphic acid G, together with fifteen known triterpenoids, was isolated from the aerial parts of G. longituba. Our results showed that euscaphic acid G exerted strong anti-proliferative activity against NCI-H460 cells in a concentration- and time-dependent manner. Flow cytometry demonstrated euscaphic acid G arrested the cell cycle at G1 phase, induced cellular apoptosis, accompanied by ROS generation and mitochondrial membrane potential reduction. Mechanistic studies revealed that euscaphic acid G treatment inhibited IKKα/β phosphorylation and IκBα phosphorylation, which subsequently caused the blockage of NF-κB p65 phosphorylation and nuclear translocation.
    CONCLUSION: In conclusion, these results suggested that euscaphic acid G from G. longituba showed potential anticancer effects against lung cancer cells via inducing cell cycle arrest and apoptosis, at least partly, through NF-κB signaling pathways.
    Keywords:  Anticancer; Glechoma longituba; Hexacyclic triterpene acid; Lung cancer; Nuclear factor-κB
    DOI:  https://doi.org/10.1016/j.phymed.2019.153037
  52. J Clin Med. 2019 Jul 31. pii: E1138. [Epub ahead of print]8(8):
    Do MH, Lee JH, Cho K, Kang MC, Subedi L, Parveen A, Kim SY.
      Lespedeza bicolor (LB) is often used in traditional medicine to remove toxins, replenish energy stores, and regulate various symptoms of diabetes. This study aimed to explore the use of LB as a therapeutic to prevent diabetic nephropathy in methylglyoxal (MGO)-treated models in vitro and in vivo. Western blotting, immunostaining, and biochemical assays were used to obtain several experimental readouts in renal epithelial cells (LLC-PK1) and BALB/c mice. These include: production of reactive oxygen species (ROS), formation of advanced glycation end-products (AGEs), expression of receptor for advanced glycation end-products (RAGE), apoptotic cell death, glucose levels, fatty acid and triglyceride levels, expression of pro-inflammatory cytokines IL-1β and TNF-α, glyoxalase 1 (Glo1), and nuclear factor erythroid 2-related factor 2 (Nrf2). Pretreatment with LB significantly reduced MGO-induced cellular apoptosis, intracellular production of ROS, and formation of AGEs to ameliorate renal dysfunction in vitro and in vivo. Interestingly, administering LB in MGO-treated cells and mice upregulated the expression of Nrf2 and Glo1, and downregulated the expression of IL-1β and TNF-α. Moreover, LB reduced MGO-induced AGE accumulation and RAGE expression in the kidneys, which subsequently reduced AGE-RAGE interactions. Overall, LB ameliorates renal cell apoptosis and corrects renal dysfunction in MGO-treated mice. These findings extend our understanding of the pathogenic mechanism of MGO-induced nephrotoxicity and regulation of the AGE/RAGE axis by Lespedeza bicolor.
    Keywords:  Lespedeza bicolor; advanced glycation end-products; diabetic nephropathy; hyperglycemia; methylglyoxal
    DOI:  https://doi.org/10.3390/jcm8081138
  53. Cancer Cell Int. 2019 ;19 188
    Zhang Y, Han CY, Duan FG, Fan XX, Yao XJ, Parks RJ, Tang YJ, Wang MF, Liu L, Tsang BK, Leung EL.
      Background: Non-small cell lung cancer (NSCLC) is the leading cause of cancer deaths primarily due to chemoresistance. Somatic mutation of TP53 (36%) and epidermal growth factor receptor (EGFR; > 30%) are major contributors to cisplatin (CDDP) resistance. Substantial evidence suggests the elevated levels of reactive oxygen species (ROS) is a key determinant in cancer. The elevated ROS can affect the cellular responses to chemotherapeutic treatments. Although the role of EGFR in PI3K/Akt signaling cascade in NSCLC is extensively studied, the molecular link between EGFR and p53 and the role of ROS in pathogenesis of NSCLC are limitedly addressed. In this study, we investigated the role of p53 in regulation of ROS production and EGFR signaling, and the chemosensitivity of NSCLC.Methods: In multiple NSCLC cell lines with varied p53 and EGFR status, we compared and examined the protein contents involved in EGFR-Akt-P53 signaling loop (EGFR, P-EGFR, Akt, P-Akt, p53, P-p53) by Western blot. Apoptosis was determined based on nuclear morphological assessment using Hoechst 33258 staining. Cellular ROS levels were measured by dichlorofluorescin diacetate (DCFDA) staining followed by flow cytometry analysis.
    Results: We have demonstrated for the first time that activation of p53 sensitizes chemoresistant NSCLC cells to CDDP by down-regulating EGFR signaling pathway and promoting intracellular ROS production. Likewise, blocking EGFR/PI3K/AKT signaling with PI3K inhibitor elicited a similar response. Our findings suggest that CDDP-induced apoptosis in chemosensitive NSCLC cells involves p53 activation, leading to suppressed EGFR signaling and ROS production. In contrast, in chemoresistant NSCLC, activated Akt promotes EGFR signaling by the positive feedback loop and suppresses CDDP-induced ROS production and apoptosis.
    Conclusion: Collectively, our study reveals that the interaction of the p53 and Akt feedback loops determine the fate of NSCLC cells and their CDDP sensitivity.
    Keywords:  Chemoresistance; Cis-diaminedichloroplatinum (Cisplatin) (CDDP); Epidermal growth factor (EGFR); Non-small cell lung cancer (NSCLC); p53 reactive oxygen species (ROS)
    DOI:  https://doi.org/10.1186/s12935-019-0910-2
  54. Mar Drugs. 2019 Aug 01. pii: E451. [Epub ahead of print]17(8):
    Rodríguez-Luna A, Ávila-Román J, Oliveira H, Motilva V, Talero E.
      Excessive exposure to ultraviolet (UV) radiation is the main risk factor to develop skin pathologies or cancer because it encourages oxidative condition and skin inflammation. In this sense, strategies for its prevention are currently being evaluated. Natural products such as carotenoids or polyphenols, which are abundant in the marine environment, have been used in the prevention of oxidative stress due to their demonstrated antioxidant activities. Nevertheless, the anti-inflammatory activity and its implication in photo-prevention have not been extensively studied. Thus, we aimed to evaluate the combination of fucoxanthin (FX) and rosmarinic acid (RA) on cell viability, apoptosis induction, inflammasome regulation, and anti-oxidative response activation in UVB-irradiated HaCaT keratinocytes. We demonstrated for the first time that the combination of FX and RA (5 µM RA plus 5 μM FX, designated as M2) improved antioxidant and anti-inflammatory profiles in comparison to compounds assayed individually, by reducing UVB-induced apoptosis and the consequent ROS production. Furthermore, the M2 combination modulated the inflammatory response through down-regulation of inflammasome components such as NLRP3, ASC, and Caspase-1, and the interleukin (IL)-1β production. In addition, Nrf2 and HO-1 antioxidant genes expression increased in UVB-exposed HaCaT cells pre-treated with M2. These results suggest that this combination of natural products exerts photo-protective effects by down-regulating NRLP3-inflammasome and increasing Nrf2 signalling pathway.
    Keywords:  NRLP3; UVB; anti-inflammatory; anti-oxidative; fucoxanthin; inflammasome; photo-protection; rosmarinic acid
    DOI:  https://doi.org/10.3390/md17080451
  55. Evid Based Complement Alternat Med. 2019 ;2019 5789574
    Yuan W, Chang H, Liu X, Wang S, Liu H, Xuan H.
      Propolis has been widely used as a dietary supplement for its health benefits, including cardiovascular protective effects. The aim of this study was to investigate the cytoprotective effects of Brazilian green propolis (BP) against oxidized low-density lipoprotein (Ox-LDL) induced human umbilical vein endothelial cells (HUVECs) damage. Our results suggested that treatment with BP rescued Ox-LDL-stimulated HUVECs cell viability losses, which might be associated with its inhibitive effects on the cell apoptosis and autophagy. We also noticed that BP restored Ox-LDL-stimulated HUVECs oxidative stress, by induced antioxidant gene expressions, including Heme oxygenase-1 and its upstream mediator, Nrf2, which were mediated by the activation of the phosphorylation of PI3K/Akt/mTOR. Pretreatment with wortmannin, PI3K/AKT inhibitor, abolished BP induced Nrf2 nuclear translocation and HO-1 level. Our results demonstrated that BP protected HUVECs against oxidative damage partly via PI3K/Akt/mTOR-mediated Nrf/HO-1 pathway, which might be applied into preventing Ox-LDL mediated cardiovascular diseases.
    DOI:  https://doi.org/10.1155/2019/5789574
  56. Sci Rep. 2019 Jul 30. 9(1): 11064
    I El-Gogary R, Gaber SAA, Nasr M.
      Baicalin is a multi-purpose flavonoid known for its anticancer properties, but its application is hindered by its low water solubility and bioavailability. Polymeric nanocapsules were proposed in this work as a promising system for enhancing baicalin delivery, and potentiating its anticancer properties. The characterization of nanocapsules was augmented with chemometric analysis, and the selected formulations were tested on two breast cancer cell lines (MCF-7 and MDA-MB-231), with mechanistic anticancer elucidation using MTT assay, confocal microscopy uptake, flow cytometry, mechanism of cell death, reactive oxygen species production, caspase 3/7 activity and death biomarker expression using quantitative real time PCR. Results showed that baicalin nanocapsules displayed favorable pharmaceutical properties; with the formulation variables affecting their properties elucidated using chemometric factorial analysis. Nanocapsules enhanced the anticancer activity of baicalin up to 216 times for MCF-7 cells and 31 times for MDA-MB-231 after 24 hr incubation. Cellular internalization of the fluorescently labeled nanocapsules was confirmed after 4 hr incubation for both cell lines. Apoptosis was the dominant cell death mechanism, with significant up-regulation of P53 in baicalin nanocapsules treated cells. Data here presented drive to further preclinical studies to investigate the delivery of baicalin polymeric nanocapsules and their anti-cancer activity.
    DOI:  https://doi.org/10.1038/s41598-019-47586-7
  57. J Cancer Prev. 2019 Jun;24(2): 65-71
    Kim Y, Jang HH.
      Peroxiredoxins (Prxs) are antioxidant enzymes that protect cells from oxidative stress by reducing intracellular accumulation of reactive oxygen species (ROS). In mammalian cells, the six Prx isoforms are ubiquitously expressed in diverse intracellular locations. They are involved in the regulation of various physiological processes including cell growth, differentiation, apoptosis, immune response and metabolism as well as intracellular ROS homeostasis. Although there are increasing evidences that Prxs are involved in carcinogenesis of many cancers, their role in cancer is controversial. The ROS levels in cancer cells are increased compared to normal cells, thus promoting cancer development. Nevertheless, for various cancer types, an overexpression of Prxs has been found to be associated with poor patient prognosis, and an increasing number of studies have reported that tumorigenesis is either facilitated or inhibited by regulation of cancer-associated signaling pathways. This review summarizes Prx isoforms and their basic functions, the relationship between the expression level and the physiological role of Prxs in cancer cells, and their roles in regulating cancer-associated signaling pathways.
    Keywords:  Cancer; Oxidative stress; Peroxidase activity; Peroxiredoxins; Tumorigenesis
    DOI:  https://doi.org/10.15430/JCP.2019.24.2.65
  58. Transplant Proc. 2019 Jul 24. pii: S0041-1345(19)30274-X. [Epub ahead of print]
    Yan Y, Lv X, Ma J, Hong G, Li S, Shen J, Chen H, Cao K, Chen S, Cheng T, Dong C, Han J, Ma H, Wu M, Wang X, Xing C, Zhu Y, Shen L, Wang Y, Tong F, Wang Z.
      PURPOSE: The objective of this research was to survey the therapeutic action of simvastatin (Sim) on intestinal ischemia/reperfusion injury (II/RI) by modulating Omi/HtrA2 signaling pathways.METHODS: Sprague Dawley rats were pretreated with 40 mg/kg Sim and then subjected to 1 hour of ischemia and 3 hours of reperfusion. The blood and intestinal tissues were collected, pathologic injury was observed, the contents of serum tumor necrosis factor-α and interleukin-6 (IL-6) were estimated, and superoxide dismutase, methane dicarboxylic aldehyde, and cysteinyl aspartate specific proteinase-3 (caspase-3) levels, as well as the expressions of Omi/HtrA2 and caspase-3, were measured in the intestinal tissues.
    RESULTS: Sim preconditioning mitigated the damnification of intestinal tissues by decreasing oxidative stress, inflammatory damage, and apoptosis and downregulating the expression of Omi/HtrA2 compared to the ischemia/reperfusion group, while Sim+Ucf-101 significantly augmented this effect.
    CONCLUSION: These results suggest that Sim may alleviate intestinal ischemia/reperfusion injury by modulating Omi/HtrA2 signaling pathways.
    DOI:  https://doi.org/10.1016/j.transproceed.2019.04.076
  59. Environ Sci Pollut Res Int. 2019 Aug 02.
    Arisha AH, Ahmed MM, Kamel MA, Attia YA, Hussein MMA.
      Silver nanoparticles (AgNPs) have been widely produced for different industrial purposes. Recently, biogenic synthesis of AgNPs has emerged although the extent of effects from exposure, oral exposure in particular, to nanomaterials synthesized in such a manner remains elusive. The main objective of this study was to evaluate the effects of oral administration of a dose of 50 mg/Kg body weight AgNPs biosynthesized in baker's yeast (Saccharomyces cerevisiae) over a period of eight weeks on the reproductive performance and the possibility of a protective effect through co-administration of morin. Forty-eight male Sprague-Dawley rats were used in four experimental groups (control, morin-treated group, AgNP-treated, and AgNP + morin co-treatment). AgNPs produced no significant alteration in daily food intake or body weight. Both the absolute and relative testicular weights were significantly reduced but not the epididymal weight. Also, serum levels of urea, creatinine, uric acid, and liver enzymes were significantly elevated. Furthermore, AgNPs significantly downregulated the hypothalamic-pituitary-gonadal axis. This corresponds to lower motility and viability percent, reduced sperm concentration, and a higher abnormality ratio as well as a prominent alteration in the blood-testis barrier (BTB) and testicular histology and induction of testicular apoptosis and oxidative stress. The supplementation of morin evidently restored most of the reproductive characters to its physiological range. We can conclude that exposure to the biologically synthesized AgNPs for an extended period of time has proven to be a health risk that can be ameliorated via oral administration of some bioactive agents including morin.
    Keywords:  Apoptosis; Blood–testis barrier; Morin; Oxidative stress; Reproduction; Silver nanoparticles
    DOI:  https://doi.org/10.1007/s11356-019-06066-1
  60. Int J Biol Sci. 2019 ;15(8): 1676-1684
    Cao P, Xia Y, He W, Zhang T, Hong L, Zheng P, Shen X, Liang G, Cui R, Zou P.
      Colon cancer is a malignant type of cancer with high prevalence and is one of the primary causes of cancer-related deaths. Oxaliplatin plays a significant role in the treatment of cancer, but the application of oxaliplatin is restricted due to its toxic side effects and drug resistance in clinical practice. Therefore, there is an urgent need for new strategies that can synergize with oxaliplatin for confronting colon cancer. Alantolactone (ALT), a natural sesquiterpene lactone, possesses antitumor properties in a number of cancer cell lines. In the present study, we investigated how ALT acts synergistically with oxaliplatin on human colorectal cancer HCT116 and RKO cells in vitro and in vivo. We observed that ALT strengthened the effect of oxaliplatin-induced growth restrain and apoptosis in HCT116 and RKO cells. It is through a mechanism concerning remarkable accumulation of intracellular reactive oxygen species (ROS) and activation of JNK and p38 MAPK signaling pathways. These changes ultimately induced apoptosis of HCT116 and RKO cells. Pretreatment of cells with the ROS reversal agent NAC significantly blocked the apoptosis induced by the combination treatment, and suppressed expression of JNK and p38 phosphorylation in HCT116 and RKO cells. In the xenograft model, the combination therapy displayed stronger antitumor activity compared with single agents. Immunohistochemistry of subsequent treatment tumors showed a significant decrease in proliferation as compared to either of the treatments alone. These results suggest that the combination treatment with ALT and oxaliplatin may become a potential therapeutic strategy for colon cancer.
    Keywords:  Alantolactone; Colon cancer; MAPK; Oxaliplatin; Reactive oxygen species
    DOI:  https://doi.org/10.7150/ijbs.35265
  61. Toxicol In Vitro. 2019 Jul 26. pii: S0887-2333(19)30299-1. [Epub ahead of print] 104592
    Miao Y, Sun X, Gao G, Jia X, Wu H, Chen Y, Huang L.
      (-)-epigallocatechin-3-gallate (EGCG), the main component of green tea, has long been explored in the treatment and/or prevention of central nervous system (CNS) disorders. However, EGCG has been recently shown to exhibit acute and subacute toxicity. Although a lot of work has been done, the mechanisms of EGCG-induced mitochondrial dysfunction has not been delineated in primary astrocyte. Here, the mitotoxic effect of EGCG on primary astrocytes was investigated by measuring Ca2+ overloading-induced mitochondrial dysfunction. As expected, EGCG dose-dependently inhibited astrocytes growth depending on Ca2+ overloading, especially at 50 μM EGCG group. It is interesting to note that Ca2+ influx from the extracellular space was responsible for an increase in the cytosolic Ca2+ level ([Ca2+]i) by opening voltage-gated calcium channels (VGCCs) and, consequently, mitochondrial Ca2+ ([Ca2+]m) overloaded via the mitochondrial Ca2+ uniporter (MCU). As a result, mitochondrial dysfunction was induced, including the opening of the mitochondrial permeability transition pore (mPTP), mitochondrial membrane depolarization, an increasing in reactive oxygen species (ROS), and cytochrosome c (cyt c) releasing. Therefore, more apoptotic cells were observed in 50 μM EGCG group than that of in 1 μM EGCG group. These findings suggested that a high dose of EGCG was toxic to astrocytes partly by targeting mitochondria via calcium pathway, which would extend our understanding of the toxicity of EGCG and the underlying mechanisms.
    Keywords:  (-)-epigallocatechin-3-O-gallate; Apoptosis; Astrocyte; Calcium; Mitochondria; Reactive oxygen species
    DOI:  https://doi.org/10.1016/j.tiv.2019.104592
  62. Nanomedicine (Lond). 2019 Jul 29.
    Kirwale S, Pooladanda V, Thatikonda S, Murugappan S, Khurana A, Godugu C.
      Aim: Selenium nanoparticles (SeNPs) may have a potential role in treating dermal disorders due to its wide therapeutic properties, but there is a need to evaluate its toxicity in keratinocytes. The present study evaluated the molecular mechanism and mode of cell death induced by SeNPs on dermal keratinocytes. Materials & methods: SeNPs were synthesized, characterized and studied in human keratinocytes cells. Oxidative stress and mitochondrial membrane depolarization were evaluated by various techniques. Additionally, autophagy mediated apoptotic cell death was evaluated. Results: SeNPs induced oxidative stress and apoptotic cell death in keratinocytes by increasing autophagy through the formation of acidic lysosomes and autophagosomes. Conclusion: Overall, SeNPs induce the oxidative stress and autophagy mediated apoptotic cell death in human keratinocytes cells.
    Keywords:  SeNPs; apoptosis; autophagy; cell death; keratinocytes; oxidative stress
    DOI:  https://doi.org/10.2217/nnm-2018-0397
  63. Int J Mol Sci. 2019 Jul 29. pii: E3702. [Epub ahead of print]20(15):
    Zakharova IO, Sokolova TV, Bayunova LV, Zorina II, Rychkova MP, Shpakov AO, Avrova NF.
      Insulin is a promising drug for the treatment of diseases associated with brain damage. However, the mechanism of its neuroprotective action is far from being understood. Our aim was to study the insulin-induced protection of cortical neurons in oxidative stress and its mechanism. Immunoblotting, flow cytometry, colorimetric, and fluorometric techniques were used. The insulin neuroprotection was shown to depend on insulin concentration in the nanomolar range. Insulin decreased the reactive oxygen species formation in neurons. The insulin-induced modulation of various protein kinase activities was studied at eight time-points after neuronal exposure to prooxidant (hydrogen peroxide). In prooxidant-exposed neurons, insulin increased the phosphorylation of GSK-3beta at Ser9 (thus inactivating it), which resulted from Akt activation. Insulin activated ERK1/2 in neurons 5-30 min after cell exposure to prooxidant. Hydrogen peroxide markedly activated AMPK, while it was for the first time shown that insulin inhibited it in neurons at periods of the most pronounced activation by prooxidant. Insulin normalized Bax/Bcl-2 ratio and mitochondrial membrane potential in neurons in oxidative stress. The inhibitors of the PI3K/Akt and MEK1/2/ERK1/2 signaling pathways and the AMPK activator reduced the neuroprotective effect of insulin. Thus, the protective action of insulin on cortical neurons in oxidative stress appear to be realized to a large extent through activation of Akt and ERK1/2, GSK-3beta inactivation, and inhibition of AMPK activity increased by neuronal exposure to prooxidant.
    Keywords:  Bax/Bcl-2 ratio; cortical neurons; insulin; mitochondrial membrane potential; oxidative stress; protection; protein kinases
    DOI:  https://doi.org/10.3390/ijms20153702
  64. Cells. 2019 Jul 30. pii: E795. [Epub ahead of print]8(8):
    Demine S, Renard P, Arnould T.
      Mitochondrial uncoupling can be defined as a dissociation between mitochondrial membrane potential generation and its use for mitochondria-dependent ATP synthesis. Although this process was originally considered a mitochondrial dysfunction, the identification of UCP-1 as an endogenous physiological uncoupling protein suggests that the process could be involved in many other biological processes. In this review, we first compare the mitochondrial uncoupling agents available in term of mechanistic and non-specific effects. Proteins regulating mitochondrial uncoupling, as well as chemical compounds with uncoupling properties are discussed. Second, we summarize the most recent findings linking mitochondrial uncoupling and other cellular or biological processes, such as bulk and specific autophagy, reactive oxygen species production, protein secretion, cell death, physical exercise, metabolic adaptations in adipose tissue, and cell signaling. Finally, we show how mitochondrial uncoupling could be used to treat several human diseases, such as obesity, cardiovascular diseases, or neurological disorders.
    Keywords:  adipocyte browning; apoptosis; autophagy; cell death; cell signaling; mitochondrial uncoupling; oxidative stress; physical exercise; protein secretion; uncoupler
    DOI:  https://doi.org/10.3390/cells8080795
  65. Antioxidants (Basel). 2019 Aug 01. pii: E261. [Epub ahead of print]8(8):
    Kushairi N, Phan CW, Sabaratnam V, David P, Naidu M.
      Oxidative stress and inflammation in neuron-glia system are key factors in the pathogenesis of neurodegenerative diseases. As synthetic drugs may cause side effects, natural products have gained recognition for the prevention or management of diseases. In this study, hot water (HE-HWA) and ethanolic (HE-ETH) extracts of the basidiocarps of Hericium erinaceus mushroom were investigated for their neuroprotective and anti-inflammatory activities against hydrogen peroxide (H2O2)-induced neurotoxicity in HT22 mouse hippocampal neurons and lipopolysaccharide (LPS)-induced BV2 microglial activation respectively. HE-ETH showed potent neuroprotective activity by significantly (p < 0.0001) increasing the viability of H2O2-treated neurons. This was accompanied by significant reduction in reactive oxygen species (ROS) (p < 0.05) and improvement of the antioxidant enzyme catalase (CAT) (p < 0.05) and glutathione (GSH) content (p < 0.01). Besides, HE-ETH significantly improved mitochondrial membrane potential (MMP) (p < 0.05) and ATP production (p < 0.0001) while reducing mitochondrial toxicity (p < 0.001), Bcl-2-associated X (Bax) gene expression (p < 0.05) and nuclear apoptosis (p < 0.0001). However, gene expression of Nuclear factor erythroid 2-related factor 2 (Nrf2), heme oxygenase 1 (HO-1) and NAD(P)H quinone dehydrogenase 1 (NQO1) were unaffected (p > 0.05). HE-ETH also significantly (p < 0.0001) reduced nitric oxide (NO) level in LPS-treated BV2 indicating an anti-inflammatory activity in the microglia. These findings demonstrated HE-ETH maybe a potential neuroprotective and anti-inflammatory agent in neuron-glia environment.
    Keywords:  Hericium erinaceus; anti-inflammation; antioxidants; mushroom; neuroprotection
    DOI:  https://doi.org/10.3390/antiox8080261
  66. Biomed Pharmacother. 2019 Jul 23. pii: S0753-3322(19)32955-5. [Epub ahead of print]118 109249
    Lin Y, Jiang M, Chen W, Zhao T, Wei Y.
      The endoplasmic reticulum (ER) acts as a moving organelle with many important cellular functions. As the ER lacks sufficient nutrients under pathological conditions leading to uncontrolled protein synthesis, aggregation of unfolded/misfolded proteins in the ER lumen causes the unfolded protein response (UPR) to be activated. Chronic ER stress produces endogenous or exogenous damage to cells and activates UPR, which leads to impaired intracellular calcium and redox homeostasis. The UPR is capable of recognizing the accumulation of unfolded proteins in the ER. The protein response enhances the ability of the ER to fold proteins and causes apoptosis when the function of the ER fails to return to normal. In different malignancies, ER stress can effectively induce the occurrence of autophagy in cells because malignant tumor cells need to re-use their organelles to maintain growth. Autophagy simultaneously counteracts ER stress-induced ER expansion and has the effect of enhancing cell viability and non-apoptotic death. Oxidative stress also affects mitochondrial function of important proteins through protein overload. Mitochondrial reactive oxygen species (ROS) are produced by calcium-enhanced ER release. The accumulation of toxic substances in ER and mitochondria in mitochondria destroys basic organelle function. It is known that sustained ER stress can also trigger an inflammatory response through the UPR pathway. Inflammatory response is thought to be associated with tumor development. This review discusses the emerging links between UPR responses and autophagy, oxidative stress, and inflammatory response signals in ER stress, as well as the potential development of targeting this multifaceted signaling pathway in various cancers.
    Keywords:  Autophagy; Cancer; Endoplasmic reticulum stress; Inflammatory; Oxidative stress
    DOI:  https://doi.org/10.1016/j.biopha.2019.109249
  67. Med Sci Monit. 2019 Jul 30. 25 5648-5656
    Li Y, Meng R.
      BACKGROUND Endothelial cells are involved in vascular homeostasis, and endothelial cell dysfunction is involved in the pathogenesis of cardiovascular disease. This study aimed to investigate the effects of microRNA-154 in human umbilical vein endothelial cells (HUVECs) following injury induced by hydrogen peroxide (H₂O₂). MATERIAL AND METHODS Cell viability and apoptosis of HUVECs treated with H₂O₂ were measured. The expression of microRNA-154 was detected by quantitative real-time polymerase chain reaction (qRT-PCR). Cell survival, caspase-3 activity, and the apoptosis rate were evaluated in H₂O₂-treated HUVECs cells after the upregulation and down-regulation of microRNA-154 expression. The interaction between microRNA-154 and Dickkopf WNT signaling pathway inhibitor 2 (DKK2) was predicted by bioinformatics analysis and was verified by luciferase reporter gene assay and Western blot. The effects of DKK2 short-interfering RNA (siRNA) on antioxidant injury in HUVECs cells were determined. RESULTS The survival rate of HUVECs exposed to H₂O₂ was significantly reduced and the apoptosis rate was significantly increased, and H₂O₂ significantly inhibited the expression of microRNA-154 in a dose-dependent manner. Overexpression of microRNA-154 increased cell survival, reduced the activity of caspase-3, and reduced cell apoptosis. Inhibition of microRNA-154 expression decreased cell survival, increased the activity of caspase-3, and promoted cell apoptosis. Luciferase reporter gene assay and Western blot showed that microRNA-154 interacted with the Wnt pathway molecule DKK2 in HUVECS. Also, DDK2 siRNA resulted in a similar protective effect on H₂O₂-treated HUVECs as overexpression of microRNA-154. CONCLUSIONS Oxidative injury in HUVECs was regulated by microRNA-154 targeting the Wnt/ß-catenin signaling pathway.
    DOI:  https://doi.org/10.12659/MSM.915263
  68. J Cardiovasc Transl Res. 2019 Jul 29.
    Fan X, Wang E, He J, Zhang L, Zeng X, Gui Y, Sun Q, Song Y, Yuan H.
      Ligustrazine is one of the alkaloid compounds isolated from the traditional Chinese herb, which shows protective effects on cardiovascular disorders. High homocysteine (Hcy) level can predict cardiovascular-related events including death. In this study, we used Hcy to stimulate the human umbilical vein endothelial cells (HUVECs) and investigated the protective effect of ligustrazine on endothelial dysfunction by assessing the cell apoptosis, oxidative damage, mitochondrial dysfunction, and the potential molecular pathways. Our results clearly showed that ligustrazine increased HUVEC cell viability, decreased the dehydrogenase (LDH) level, and inhibited HUVEC apoptosis, which was associated with the attenuation of attenuated oxidative damage. The mitochondrial-dependent pathway was closely related in the regulation of ligustrazine, reflected by the attenuated mitochondrial membrane potential change and decreased cytochrome c release from the mitochondria to the cytosol. Ligustrazine may protect Hcy-induced apoptosis in HUVECs by attenuating oxidative damage and modulating mitochondrial dysfunction.
    Keywords:  Cell apoptosis; Homocysteine; Human umbilical vein endothelial cells (HUVECs); Ligustrazine; Mitochondria dysfunction
    DOI:  https://doi.org/10.1007/s12265-019-09900-6
  69. Oxid Med Cell Longev. 2019 ;2019 6325424
    de Lima-Seolin BG, Nemec-Bakk A, Forsyth H, Kirk S, da Rosa Araujo AS, Schenkel PC, Belló-Klein A, Khaper N.
      The increased circulation of norepinephrine, found in the diseased heart as a result of sympathetic nervous system overactivation, is responsible for its cardiotoxic effects including pathological hypertrophy, cell death, and oxidative stress. Bucindolol is a third generation adrenergic blocker, which acts on the β1 and β2 receptors, and has additional α1 antagonist activity. Thus, the aim of this study was to investigate the action of bucindolol on oxidative stress, hypertrophy, cell survival, and cell death signaling pathways in H9c2 cardiac cells exposed to norepinephrine. H9c2 cells were incubated with 10 μM norepinephrine for 24 h in the presence or absence of bucindolol (10 μM) treatment for 8 h. Western blot was used to determine the expression of proteins for hypertrophy/survival and death signaling pathways. Flow cytometry was used to assess cell death via caspase-3/7 activity and propidium iodide and reactive oxygen species via measuring the fluorescence of CM-H2DCFDA. Norepinephrine exposure resulted in an increase in oxidative stress as well as cell death. This was accompanied by an increased protein expression of LC3B-II/I. The protein kinase B/mammalian target of the rapamycin (Akt/mTOR) pathway which is involved in cardiac remodeling process was activated in response to norepinephrine and was mitigated by bucindolol. In conclusion, bucindolol was able to modulate cardiac remodeling which is mediated by oxidative stress.
    DOI:  https://doi.org/10.1155/2019/6325424
  70. Int Immunopharmacol. 2019 Jul 25. pii: S1567-5769(19)30468-0. [Epub ahead of print]75 105697
    Zhao F, Dang Y, Zhang R, Jing G, Liang W, Xie L, Li Z.
      Acrylonitrile (ACN) is often found in the productions of synthetic fibers, rubber, and plastics. Exposure to ACN could cause pathological changes of the nervous system, which appeared early and were very serious. Current studies have found that the neurotoxicity is mainly related to oxidative damage and inflammation induced by ACN. Apigenin (AP) is a flavonoid subtype compound that is less toxic, non-mutagenic, and widely distributed in many types of vegetables and fruits. Studies have confirmed that it has nice antioxidant, anti-inflammatory and anti-apoptotic properties in the nervous system and related disease models, such as Alzheimer's disease. In this study, we used AP (117, 234 and 351 mg·kg-1) pretreatment intragastrically to resist the neurotoxicity caused by ACN gavage (46 mg·kg-1) for 28 days, and then detected the oxidative stress, inflammation mediated by the TLR4/NF-κB signaling pathway, and apoptosis to evaluate the protective effect of AP. The results showed that AP could lessen the autonomic activities of rats, and improve the abnormal morphology of neurons induced by ACN. AP could also reduce the oxidative stress, downregulate the TLR4/NF-κB signaling pathway, decrease the levels of interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α), and inhibit the mitochondria-mediated neuron apoptosis. Immunofluorescence result showed that AP could decrease the activation and nuclear transfer of NF-κB induced by ACN. These results suggested that AP could protect the brain against ACN-induced neurotoxicity by inhibiting the TLR4/NF-κB signaling pathway and could exhibit a neuroprotective effect.
    Keywords:  Acrylonitrile (ACN); Apigenin (AP); Inflammation; Neuroprotection; Neurotoxicity
    DOI:  https://doi.org/10.1016/j.intimp.2019.105697
  71. Biosci Biotechnol Biochem. 2019 Jul 31. 1-7
    Lee JW, Lee S, Ho JN, Youn JI, Byun SS, Lee E.
      As standard second-line regimen has not been established for patients who are refractory to or relapse with cisplatin-based chemotherapy, an effective class of novel chemotherapeutic agents is needed for cisplatin-resistant bladder cancer. Recent publications reported that MutT homolog 1 (MTH1) inhibitors suppress tumor growth and induce impressive therapeutic responses in a variety of human cancer cells. Few studies investigated the cytotoxic effects of MTH1 inhibitors in human bladder cancer. Accordingly, we investigated the antitumor effects and the possible molecular mechanisms of MTH1 inhibitors in cisplatin-sensitive (T24) and - resistant (T24R2) human bladder cancer cell lines. These results suggest that TH588 or TH287 may induce cancer cell suppression by off-target effects such as alterations in the expression of apoptosis- and cell cycle-related proteins rather than MTH1 inhibition in cisplatin-sensitive and - resistant bladder cancer cells. Abbreviations: MTH: MutT homolog; ROS: reactive oxygen species; CCK-8: cell counting kit-8; DCFH-DA: dichlorofluorescein diacetate; PARP: poly (ADP-ribose) polymerase.
    Keywords:  Bladder cancer; MutT homolog 1; antitumor effect; cisplatin; drug resistance
    DOI:  https://doi.org/10.1080/09168451.2019.1648207
  72. Int J Immunopathol Pharmacol. 2019 Jan-Dec;33:33 2058738419866021
    Jia F, Mou L, Ge H.
      Ginsenoside Rb1 (Rb1) possesses a cardioprotective effect via mediating microRNAs (miRs), while it is unexplored whether miR-210 is regulated by Rb1 in response to oxidative stress. Human endothelial EA.hy926 cells were stimulated with H2O2 before Rb1 treatment. After transfection, cell viability, apoptosis, migration, and invasion assays were conducted. Western blot was applied to quantify protein. BCL2/adenovirus E1B 19-kDa interacting protein 3 (BNIP3) and miR-210 were analyzed with quantitative reverse transcription polymerase chain reaction. Dual luciferase activity assay was performed. Rb1 elevated viability, migration, and invasion of H2O2-treated cells. H2O2-induced apoptosis was moderated by Rb1. miR-210 was augmented in H2O2-treated cells after Rb1 stimulation. miR-210 inhibitor abolished the positive effects of Rb1. BNIP3 was negatively modulated by miR-210 and implicated in modulating viability, apoptosis, and migration and invasion. In addition, BNIP3 modulated phosphorylation of regulators. Rb1 repressed oxidative injury via elevating miR-210. miR-210 negatively mediated BNIP3, which participated in oxidative damage via regulating mammalian targets of rapamycin (mTOR) and nuclear factor-κB (NF-κB).
    Keywords:  BNIP3; ginsenoside Rb1; miR-210; oxidative stress
    DOI:  https://doi.org/10.1177/2058738419866021
  73. Adv Exp Med Biol. 2019 ;1145 305-319
    Azad MAK, Nation RL, Velkov T, Li J.
      Polymyxin-induced nephrotoxicity is the major dose-limiting factor and can occur in up to 60% of patients after intravenous administration. This chapter reviews the latest literature on the mechanisms of polymyxin-induced nephrotoxicity and its amelioration. After filtration by glomeruli, polymyxins substantially accumulate in renal proximal tubules via receptor-mediated endocytosis mainly by megalin and PEPT2. It is believed that subsequently, a cascade of interconnected events occur, including the activation of death receptor and mitochondrial apoptotic pathways, mitochondrial damage, endoplasmic reticulum stress, oxidative stress and cell cycle arrest. The current literature shows that oxidative stress plays a key role in polymyxin-induced kidney damage. Use of antioxidants have a potential in the attenuation of polymyxin-induced nephrotoxicity, thereby widening the therapeutic window. Mechanistic findings on polymyxin-induced nephrotoxicity are critical for the optimization of their use in the clinic and the discovery of safer polymyxin-like antibiotics.
    Keywords:  Apoptosis; Cell cycle; Nephrotoxicity; Oxidative stress; Polymyxin
    DOI:  https://doi.org/10.1007/978-3-030-16373-0_18
  74. Cell Death Differ. 2019 Jul 31.
    Ma K, Zhang Z, Chang R, Cheng H, Mu C, Zhao T, Chen L, Zhang C, Luo Q, Lin J, Zhu Y, Chen Q.
      Mitochondria are highly dynamic organelles and respond to stress by changing their fission-fusion cycle, undergoing mitophagy, or releasing apoptotic proteins to initiate cell death. The molecular mechanisms that sense different stresses and coordinate distinct effectors still await full characterization. Here, we show that PGAM5, which exists in an equilibrium between dimeric and multimeric states, dephosphorylates BCL-xL to inhibit apoptosis or FUNDC1 to activate mitofission and mitophagy in response to distinct stresses. In vinblastine-treated cells, PGAM5 dephosphorylates BCL-xL at Ser62 to restore BCL-xL sequestration of BAX and BAK and thereby resistance to apoptosis. Selenite-induced oxidative stress increases the multimerization of PGAM5, resulting in its dissociation from BCL-xL, which causes increased BCL-xL phosphorylation and apoptosis. Once freed, the more multimeric and active PGAM5 dephosphorylates FUNDC1 to initiate mitofission and mitophagy. The reciprocal interaction of PGAM5 with FUNDC1 and BCL-xL, controlled by PGAM5 multimerization, serves as a molecular switch between mitofission/mitophagy and apoptosis.
    DOI:  https://doi.org/10.1038/s41418-019-0396-4
  75. J Food Biochem. 2019 Aug 01. e13001
    El-Mansi AA, ElSayyad HI, Elshershaby EM, Al-Ashry NE.
      Our study was conducted to characterize the efficacy of barley and/or date palm fruits (10%) in alleviation of hypercholesterolemic endometrial insults in obese rat model. Sixty-four Wistar albino rats were randomized into eight groups (n = 8); control, hypercholesterolemic- and hypercholesterolemic-treated groups. Animals were subjected to treatment for 4 months. After sacrifice, serum and uterine tissues were collected and processed for biochemical, histological, immunohistochemical, and electron microscopic investigations. In hypercholesterolemic rats, the endometrium displayed hyperplasia with necrotic patches in the surface epithelium and its glandular lining cells. Also, there was a remarkable increase in the endometrial thickness and significant decrease in corresponding glandular numbers. Prompted by these findings, immunohistochemical localization revealed that expression of proliferating cell nuclear antigen was downregulated, while cleaved caspase-3 was upregulated in the endometrial cells in hypercholesterolemic group. Accordingly, there was remarkable depletion of antioxidant enzymatic activities associated with increased lipid peroxidation and apoptotic markers. Contradictory, supplementation of barley and/or dates to hypercholesterolemic groups showed intriguing amelioration for the histological architecture of the endometrium and balancing its oxidative redox. In conclusion, the administration of barley and/or dates confers enhanced synergistic effects in attenuation of hypercholesterolemic induced-endometrial dysfunction. This is clear evidence that endometrial amelioration was directly linked to the implication of highly potential antioxidant capacity of barley and/or dates phytochemicals, β-glucan, polyphenols, and other trace elements, which can be utilized to establish a phyto-therapeutic strategy for activating endometrial cell regeneration. PRACTICAL APPLICATIONS: Barley and dates confer both hypoglycemic and hypocholesterolemic potentials. Therefore, their ingredients would be implicated in the amelioration of uterine functions in obese women. These favorable potentials were directly linked to the restraining of endometrial inflammation and retrieving the oxidative capacity. Furthermore, our findings demonstrated that barley and dates substantially diminished the expression of TNF-α, mitigated DNA damage and prevented leukocytic infiltration in the endometrial tissue; based on their high content of dietary phytochemicals, β-glucan, polyphenols, and other trace elements.
    Keywords:  apoptosis; barley; dates; endometrium; hypercholesterolemia; oxidative stress
    DOI:  https://doi.org/10.1111/jfbc.13001
  76. Urol J. 2019 07 30.
    Gozukara KH, Ozcan O, Ozgur T, Kaya YS, Tutuk O.
      PURPOSE: To evaluate the short-term use of colchicine on preventing ischemia-reperfusion injury after surgery in an experimental animal model.MATERIALS AND METHODS: A total of 40 rats were divided into five groups (n = 8). Sham (Sh), ischemia-reperfusion (I/R), I/R and colchicine-treated for once per-operatively (I/Rc1), I/R and colchicine-treated for 5 days postoperatively (I/Rc5), and I/R and placebo given for 5 days (I/Rp) groups. Testicular torsion was created by rotating the testicle 720o in clockwise direction and held for 3 hours. In group I/Rc1 30 minutes before detorsion, p.o. 1 mg/kg mL infusion of colchicine was given only once. In group I/Rc5, colchicine continued p.o. once daily for five days. Tissue malonyldialdehite (MDA), superoxide dismutase (SOD), glutathione peroxidase (GPx) and catalase (CAT) were measured for evaluating the oxidative stress. Apoptosis levels shown with Caspase-3 staining and mean seminiferous tubular diameter (MSTD), germinal epithelial cell thickness (GECT), and mean testicular biopsy score (MTBS) were used to evaluate the germ cell damage.
    RESULTS: Decreased protein MDA levels therewithal increased SOD, CAT and GPx levels achieved in I/Rc5 group when compared to I/R group and did not differ from the I/Rp group (p<0.05). MSTD, GECT, and JS were better in I/Rc5 than I/Rp which showed the natural course of I/R damage in testis (p<0.005). Caspase 3 positivity, as an apoptosis indicator, were significantly lower (p<0.05) in I/Rc5 group in comparison with I/R, I/Rc1, and I/Rp groups.
    CONCLUSION: The usage of colchicine as a complementary treatment after definitive surgery reduce early-onset ischemia-reperfusion damage and diminishes apoptosis.
    DOI:  https://doi.org/10.22037/uj.v0i0.4918
  77. J Cardiovasc Pharmacol. 2019 Jul 22.
    Wu Z, Zai W, Chen W, Jin X, Liu H.
      Doxorubicin (DOX) is a representative antibiotic of terpenoids and clinically used in the treatment of various malignant tumors. However, its application is limited by the cardiotoxocity. Curdione, an extract from Rhizoma Curcumae, has many promising pharmacological effects including protecting acute liver injury and cerebral ischemia. It is still unknown whether curdione has a protective function for DOX-induced cardiotoxicity. In our study, we investigated the protective effects of curdione against DOX-induced cardiotoxicity. Our results showed that curdione attenuated DOX-induced growth inhibition and release of lactic dehydrogenase (LDH) in a concentration-dependent manner. And curdione ameliorated the histopathological damage, reduced the elevation of serum creatine kinase-MB isoenzyme (CK-MB) and LDH by DOX. Furthermore, curdione inhibited DOX-induced cell apoptosis and modulated the expression of Bcl-2 and Bax proteins, as well as abrogated DOX-induced ROS accumulation and prevented mitochondria dysfunction. Further study indicated that curdione decreased DOX-induced phosphorylation of extracellular signal regulated kinase1/2 (Erk1/2) and c-Jun-N-terminal kinase (JNK) and activated nuclear factor-erythroid 2-related factor 2/heme oxygenase 1 (Nrf2/HO-1) signal pathway. Our results suggested that curdione maybe is a new and feasible strategy to prevent DOX-induced cardiotoxicity through monitoring multiple targets.
    DOI:  https://doi.org/10.1097/FJC.0000000000000692
  78. Evid Based Complement Alternat Med. 2019 ;2019 6503752
    Dong Y, Duan L, Chen HW, Liu YM, Zhang Y, Wang J.
      Coronary heart disease (CHD) is the worldwide leading cause for cardiovascular death. Panax notoginseng saponin (PNS), which is the main bioactive compound of panax notoginseng, has been generally accepted to exert a remarkable effect on CHD for a long time. However, to reveal the underlying treatment target and corresponding mechanism of PNS against CHD is still a substantial challenge. In this work, the targets and mechanism of PNS against CHD were successfully achieved by pharmacology-based prediction and experimental verification. 36 common targets were screened out through integrating the gene expression profile of CHD and the chemical-protein data of PNS. Then, two key nodes were further selected for verification by experiment after analyzing GO function, KEGG pathway, coexpression, and topology analysis. Results showed that PNS has protected the human umbilical vein endothelial cells from H2O2-induced oxidative stress by inhibiting early cell apoptosis via upregulating VEGFA mRNA expression. Therefore, our research has successfully pointed out one treatment target and apoptotic inhibition caused by PNS with method of integrating bioinformatics prediction and experimental verification, which has partially explained the pharmacological mechanism of PNS against CHD.
    DOI:  https://doi.org/10.1155/2019/6503752
  79. Cancer Immunol Res. 2019 Jul 30. pii: canimm.0367.2018. [Epub ahead of print]
    Nath PR, Pal-Nath D, Mandal A, Cam MC, Schwartz AL, Roberts DD.
      Elevated CD47 expression in some cancers is associated with decreased survival and limited clearance by phagocytes expressing the CD47 counter-receptor SIRPα. In contrast, elevated CD47 mRNA expression in human melanomas was associated with improved survival. Gene expression data was analyzed to determine a potential mechanism for this apparent protective function and suggested that high CD47 expression increases recruitment of natural killer (NK) cells into the tumor microenvironment. The CD47 ligand thrombospondin-1 inhibited NK cell proliferation and CD69 expression in vitro. Cd47-/- NK cells correspondingly displayed augmented effector phenotypes, indicating an inhibitory function of CD47 on NK cells. Treating human NK cells with a CD47 antibody that blocks thrombospondin-1 binding abrogated its inhibitory effect on NK cell proliferation. Similarly, treating wildtype mice with a CD47 antibody that blocks thrombospondin-1 binding delayed B16 melanoma growth, associating with increased NK cell recruitment and increased granzyme B and interferon-γ levels in intratumoral NK but not CD8+ T cells. However, B16 melanomas grew faster in Cd47-/- versus wildtype mice. Melanoma-bearing Cd47-/- mice exhibited decreased splenic NK cell numbers with impaired effector protein expression and elevated exhaustion markers. Pro-apoptotic gene expression in Cd47-/- NK cells was associated with stress-mediated increases in mitochondrial proton leak, reactive oxygen species, and apoptosis. Global gene expression profiling in NK cells from tumor-bearing mice identified CD47-dependent transcriptional responses that regulate systemic NK activation and exhaustion. Therefore, CD47 positively and negatively regulates NK cell function, and therapeutic antibodies that block inhibitory CD47 signaling can enhance NK immune surveillance of melanomas.
    DOI:  https://doi.org/10.1158/2326-6066.CIR-18-0367
  80. Toxicol Sci. 2019 Jul 31. pii: kfz160. [Epub ahead of print]
    Bartosz P, Weronika K, Żaneta B, Beata S, Beata B, Beata SŚ, Alicja M, Alicja S, Julita W, Maria W, Bogusława B.
      BACKGROUND AND PURPOSE: Benzophenone-3 is the most commonly used UV filter. It is well absorbed through the skin and gastrointestinal tract. Its best-known side effect is the impact on the function of sex hormones. Little is known about the influence of BP-3 on the brain.The aim of this study was to show whether BP-3 crosses the blood-brain barrier, determine whether it induces nerve cell damage in susceptible brain structures and identify the mechanism of its action in the CNS.EXPERIMENTAL APPROACH: BP-3 was administered dermally during the prenatal period and adulthood to rats. BP-3 effect on short-term and spatial memory was determined by novel object and novel location recognition tests. BP-3 concentrations was assayed in the brain and peripheral tissues. In brain structures, selected markers of brain damage were measured.
    KEY RESULTS: The study showed that BP-3 is absorbed through the rat skin, passes through the blood-brain barrier. BP-3 raised oxidative stress and induced apoptosis in the brain. BP-3 increased the concentration of extracellular glutamate in examined brain structures and changed the expression of glutamate transporters. BP-3 had no effect on short-term memory but impaired spatial memory.
    CONCLUSIONS AND IMPLICATIONS: The present study showed that dermal BP-3 exposure may cause damage to neurons what might be associated with the increase in the level of extracellular glutamate, most likely evoked by changes in the expression of GLT-1 and xCT glutamate transporters. Thus, exposure to BP-3 may be one of the causes that increase the risk of developing neurodegenerative diseases.
    Keywords:  benzophenone-3; caspase-3; extracellular glutamate; lipid peroxidation; memory; rat
    DOI:  https://doi.org/10.1093/toxsci/kfz160
  81. Artif Cells Nanomed Biotechnol. 2019 Dec;47(1): 3129-3137
    Zhang M, Wang X, Yao J, Qiu Z.
      Endothelial cell (EC) dysfunction induces atherosclerotic coronary heart disease (CHD) development. Recent studies demonstrated that lncRNA NEAT1 mediates multiple biological functions of cells. How NEAT1 regulates EC function is still unclear, so this study explored the role and mechanism of NEAT1 in oxidative stress-induced ECs. The levels of NEAT1 and miR-181d-5p were measured in serum samples from ApoE-/- mice and t-BHP-treated human umbilical vein endothelial cells (HUVECs) by qRT-PCR. The potential role of NEAT1 in viability, migration and apoptosis was analyzed by CCK-8, cell metastasis, flow cytometry, dual-luciferase reporter, RNA immunoprecipitation and Western blot assays using HUVECs overexpressing NEAT1. The expression of NEAT1 was increased, but miR-181d-5p expression was decreased in serum samples from both ApoE-/- mice and t-BHP-treated HUVECs. Overexpression of NEAT1 increased viability, migration and CDKN3 expression but decreased apoptotic rates, caspase-3 activity and miR-181d-5p expression in HUVECs. In addition, NEAT1 acted as a promoter of the proangiogenic capacity of HUVECs by targeting miR-181d-5p/CDKN3. Altogether, these findings indicate that NEAT1 may exert a protective effect on HUVECs by regulating the miR-181d-5p/CDKN3A axis.
    Keywords:  CDKN3; NEAT1; human umbilical vein endothelial cells; miR-181d-5p
    DOI:  https://doi.org/10.1080/21691401.2019.1646264
  82. J Orthop Surg Res. 2019 Jul 29. 14(1): 241
    Ni B, Shen H, Wang W, Lu H, Jiang L.
      BACKGROUND: The aim of this study is to explore the effects of TGF-β1 on autophagy and apoptosis induced by exogenous hydrogen peroxide (H2O2) in annulus fibrosus (AF) cells and possible signal pathways involved in this process.METHODS: AF cells were isolated from rat lumbar discs and subjected to different concentrations of exogenous H2O2 (50, 100, 200 μmol/L) for different time periods (0.5, 1, 2, and 4 h). Cell viability was determined by CCK-8 assay, and the levels of autophagy and apoptosis were evaluated by Western blotting and caspase 3, 8, 9 activity assay. By administration with different concentrations of TGF-β1 (5, 10, 20 ng/mL), the effects of TGF-β1 on autophagy and apoptosis induced by H2O2 were observed, and the possible signaling pathways were also investigated by using various apoptosis inhibitors or an autophagy inhibitor Bafilomycin A (Baf A) in AF cells.
    RESULTS: H2O2 significantly impaired cell viability in a dose- and time-dependent manner. H2O2 also induced a sudden and the highest level of autophagy at 1 h, and gradually increased apoptosis through ERK pathway. The mitochondrial pathway was involved in H2O2-induced apoptosis in AF cells. TGF-β1 reduced the expression of p-ERK and downregulated the expressions of Beclin-1, LC3 II/I, and mitochondrial-related apoptotic proteins (Bax/Bcl-2, caspase-9). Meanwhile, TGF-β1 downregulated the level of intracellular H2O2 through upregulating the expression level of glutathione peroxidase-1 (GPx-1).
    CONCLUSIONS: TGF-β1 reduced autophagy and apoptosis induced by exogenous H2O2 through downregulating the expression of ERK in AF cells. TGF-β1 could downregulate the level of ERK and intracellular H2O2 by upregulating GPx-1.
    Keywords:  Annulus fibrosus cells; Apoptosis; Autophagy; ERK; Glutathion peroxidase-1; Hydrogen peroxide; Intervertebral disc degeneration
    DOI:  https://doi.org/10.1186/s13018-019-1260-4
  83. Phytother Res. 2019 Jul 30.
    Li M, Zhu Y, Li J, Chen L, Tao W, Li X, Qiu Y.
      Verbascoside is a water-soluble natural phenylethanoid glycoside and distributes widely in plants. It has been proved with antioxidant, neuroprotective, anti-inflammatory, antibacterial, and immunomodulatory bioactivities. In this experiment, the effect and mechanism of verbascoside on hypoxic memory injury were studied in a low-pressure and low-oxygen chamber. Verbascoside (50, 150, and 300 mg/kg) was intragastrically administered once a day for 7 days. On the fourth day, rats were placed in the chamber to simulate a 7,500 m high-altitude environment The eight-arm maze was used to test the memory ability. The levels of MDA, GSH, and T-SOD in plasma, brain-NH, and hippocampus were detected. The mRNA expression of mTOR, P70S6K, and 4E-BP1 in the hippocampus tissue was determined by PCR. The protein expression of P-mTOR, P-P70S6K, P-4E-BP1, and Cleaved Caspase-3 in the hippocampus tissue was determined by western blot. The results indicated that administration with verbascoside could obviously reduce the working memory error, reference memory error, total errors, and total time; relieve the neuron damage in CA1 region of the hippocampus; and decrease the oxidative stress correlation enzyme activity in plasma, brain, and hippocampus. The amelioration of verbascoside on high altitude-induced memory impairment may be associated with the adjustment of oxidative stress and mTOR signaling pathway.
    Keywords:  apoptosis; eight-arm maze; hypoxia memory impairment; mTOR signaling pathway; oxidative stress; verbascoside
    DOI:  https://doi.org/10.1002/ptr.6443
  84. Haematologica. 2019 Aug 01. pii: haematol.2018.207001. [Epub ahead of print]
    Vaikari VP, Du Y, Wu S, Zhang T, Metzeler K, Batcha AMN, Herold T, Hiddemann W, Akhtari M, Alachkar H.
      In an effort to identify target genes in acute myeloid leukemia, we compared gene expression profiles between normal and acute myeloid leukemia cells from various publicly available datasets. We identified CD99, a gene that is upregulated in patients with acute myeloid leukemia. In 186 patients from The Cancer Genome Atlas - acute myeloid leukemia dataset, CD99 was overexpressed in patients with FLT3-ITD and was downregulated in patients with TP53 mutations. CD99 is a transmembrane protein expressed on leukocytes and plays a role in cell adhesion, trans-endothelial migration and T cells differentiation. CD99 gene encodes two isoforms with distinct expression and functional profiles in both normal and malignant tissues. Here, we report that though the CD99 long isoform initially induces an increase in cell proliferation, it also induces higher levels of reactive oxygen species (ROS), DNA damage, apoptosis and subsequent decrease in cell viability. In several leukemia murine models, the CD99 long isoform delayed disease progression and resulted in lower leukemia engraftment in the bone marrow. Furthermore, the CD99 monoclonal antibody reduced cell viability, colony formation, cell migration as well as induced cell differentiation and apoptosis in leukemia cell lines and primary blasts. Mechanistically, CD99 long isoform resulted in transient induction followed by a dramatic decrease of both ERK and SRC phosphorylation. Altogether, our study provides new insights into the role of CD99 isoforms in acute myeloid leukemia that could potentially be relevant for the preclinical development of CD99 targeted therapy.
    Keywords:  Acute Myeloid Leukemia; CD99; FLT3-ITD; Hematopoietic Stem Cell; therapeutic target
    DOI:  https://doi.org/10.3324/haematol.2018.207001
  85. Biol Reprod. 2019 Jul 30. pii: ioz135. [Epub ahead of print]
    Zhang C, Shen J, Kong S, Zhang M, Zhang Q, Zhou J, Zhen X, Kang N, Jiang Y, Ding L, Sun H, Yan G.
      Oxidative stress induces granulosa cell (GC) apoptosis and subsequent follicular atresia. Since our previous studies indicate that miR-181a expression is increased in GCs undergoing apoptosis, the present study was designed to define the relationship between exposure to oxidative stressors in GCs and changes in miR-181a expression and function. To achieve this, we employed an H2O2-induced in vitro model and a 3-nitropropionic acid (3-NP)-induced in vivo model of ovarian oxidative stress. We demonstrated that in vitro miR-181a overexpression promoted GC apoptosis in a dose-dependent manner; sphingosine-1-phosphate (S1P) significantly reversed both H2O2-induced and miR-181a-induced apoptosis in GCs. Moreover, we identified sphingosine-1-phosphate receptor 1 (S1PR1), a critical receptor of S1P, as a novel target of miR-181a in GCs. miR-181a induced GC apoptosis by repressing S1PR1 expression in vitro. Importantly, increased miR-181a expression and decreased S1PR1 expression were detected in the in vivo ovarian oxidative stress model by Western blot analysis and immunohistochemistry. Furthermore, we found similar expression patterns of miR-181a and S1PR1 in GCs from patients with premature ovarian insufficiency (POI). In conclusion, our results suggest that miR-181a directly suppresses expression of S1PR1, which has critical roles in mediating oxidative stress-induced GC apoptosis both in vitro and in vivo.
    Keywords:  S1PR1; apoptosis; granulosa cell; microRNA-181a
    DOI:  https://doi.org/10.1093/biolre/ioz135
  86. Hum Exp Toxicol. 2019 Aug 01. 960327119865588
    Elkady MA, Shalaby S, Fathi F, El-Mandouh S.
      BACKGROUND: Cyclophosphamide (CP) causes premature ovarian failure (POF) due to ovarian toxicity. The toxicity mechanism is attributed to oxidative stress, inflammation, and apoptosis. We assessed whether quercetin and rosuvastatin could promote ovarian protection against CP ovotoxicity.METHODS: A total of 80 female BALB/c mice were randomly assigned; 10 mice into each of eight groups. Group 1 (control), group 2 (EH), group 3 (CP), group 4 (QH), group 5 (QL), group 6 (RH), group 7 (RL), and group 8 (COM).
    RESULTS: Quercetin and rosuvastatin groups (4:8) showed signs of restored ovarian function in the form of a significant, dose-dependent increase in primordial follicles number, serum anti-Mullerian hormone level, and ovarian tissue glutathione level (p < 0.05) versus group 3, and a significant, dose-dependent decrease in atretic follicles number and ovarian tissue level of malondialdehyde (p < 0.05) versus group 3. Immunohistochemistry analysis demonstrated a lower expression of caspase and nuclear factor-kappa B of groups (4:8) versus group 3, although quercetin and rosuvastatin showed a nonsignificant reduction in tumor volume.
    CONCLUSIONS: We demonstrated the protective effect of quercetin and rosuvastatin against ovarian toxicity and POF induced by CP without compromising its antitumor effect.
    Keywords:  Ehrlich tumor; Premature ovarian failure; cyclophosphamide; quercetin; rosuvastatin
    DOI:  https://doi.org/10.1177/0960327119865588
  87. Biochimie. 2019 Jul 28. pii: S0300-9084(19)30220-2. [Epub ahead of print]
    Zhang Q, Liu C, Li Q, Li J, Wu Y, Liu J.
      MicroRNA-25-5p (miR-25-5p) may be involved in the pathogenesis and processes of vascular diseases. The aim of this study was to investigate the role of miR-25-5p in oxidized low-density lipoprotein (ox-LDL)-treated human brain microvessel endothelial cells (HBMECs) and the underlying mechanisms. RT-qPCR and/or western blot were used to detect the expression levels of miR-25-5p and neuronal growth regulator 1 (NEGR1). The effect of miR-25-5p overexpression and NEGR1 silencing on cell proliferation, migration, apoptosis and reactive oxygen species (ROS) production of HBMECs were measured by using CCK-8 assay, transwell assay and flow cytometry, respectively. The expression levels of apoptosis-related protein (cleaved caspase-3 and pro-caspase-3) were detected using western blot, and the nitric oxide (NO) production was measured by a nitric oxide assay kit. The expression level of miR-25-5p was decreased in HBMECs treated with ox-LDL. Compared with the control group, miR-25-5p overexpression significantly promoted the proliferation and migration of HBMECs treated with ox-LDL (p<0.01). Overexpression of miR-25-5p significantly suppressed cell apoptosis, ROS production and NO reduction of ox-LDL-induced HBMECs (p<0.01). In addition, the target gene of miR-25-5p was predicted to be NEGR1 through Targetscan online analysis. The effect of NEGR1 silencing on cell proliferation, migration, apoptosis, ROS and NO production of ox-LDL-induced HBMECs was similar to that of miR-25-5p overexpression. Furthermore, miR-25-5p overexpression and NEGR1 silencing significantly downregulated the protein expression levels of JAK2 and STAT3. Thus, miR-25-5p neutralizes the effects of ox-LDL on multiple functions of HBMECs through suppressing the expression of NEGR1 via regulating the JAK/STA signaling pathway.
    Keywords:  Human brain microvessel endothelial cells; JAK/STAT signaling pathway; MicroRNA-25-5p; Oxidized low-density lipoprotein; neuronal growth regulator 1
    DOI:  https://doi.org/10.1016/j.biochi.2019.07.020
  88. Cancer Chemother Pharmacol. 2019 Jul 31.
    Mohammadi F, Soltani A, Ghahremanloo A, Javid H, Hashemy SI.
      Thioredoxin (Trx), thioredoxin reductase (TrxR), and NADPH are key members of the Trx system that is involved in redox regulation and antioxidant defense. In recent years, several researchers have provided information about the roles of the Trx system in cancer development and progression. These reports indicated that many tumor cells express high levels of Trx and TrxR, which can be responsible for drug resistance in tumorigenesis. Inhibition of the Trx system may thus contribute to cancer therapy and improving chemotherapeutic agents. There are now a number of effective natural and synthetic inhibitors with chemotherapy applications possessing antitumor activity ranging from oxidative stress induction to apoptosis. In this article, we first described the features and functions of the Trx system and then reviewed briefly its correlations with cancer. Finally, we summarized the present knowledge about the Trx/TrxR inhibitors as anticancer drugs.
    Keywords:  Cancer; Cancer therapy; Oxidative stress; Thioredoxin; Thioredoxin reductase
    DOI:  https://doi.org/10.1007/s00280-019-03912-4
  89. Oxid Med Cell Longev. 2019 ;2019 3206542
    Li R, Liu Y, Shan YG, Gao L, Wang F, Qiu CG.
      Previous studies demonstrated that Bailcalin (BAI) prevented cardiac injuries under different disease models. Whether BAI protected against type 2 diabetes mellitus- (T2DM-) associated cardiomyopathy was investigated in this study. T2DM was established by the combination of streptozotocin injection and high-fat diet in mice. BAI was administered daily for 6 months. After evaluating cardiac functions, mice hearts were removed and processed for morphological, biochemical, and molecular mechanism analyses. Neonatal rat cardiomyocytes (NRCM) were isolated and treated with high glucose and palmitate (HG/Pal) for in vitro investigation. BAI significantly ameliorated T2DM-induced cardiomyocyte hypertrophy, interstitial fibrosis, and lipid accumulation accompanied by markedly improved cardiac functions in diabetic mice. Mechanically, BAI restored decreased phosphorylation of AMPK and enhanced expression and nuclei translocation of Nrf2. In in vitro experiments, BAI also prevented NRCM from HG/Pal-induced apoptosis and oxidative stress injuries by increasing p-AMPK and Nrf2 accumulation. The means by which BAI restored p-AMPK seemed to be related to the antioxidative effects of Nrf2 after silencing AMPK or Nrf2 in NRCM. Furthermore, BAI regulated Nrf2 by inhibiting Nrf2 ubiquitination and consequent degradation mediated by Keap1. This study showed that BAI alleviated diabetes-associated cardiac dysfunction and cardiomyocyte injuries in vivo and in vitro via Keap1/Nrf2/AMPK-mediated antioxidation and lipid-lowering effects. BAI might be a potential adjuvant drug for diabetes cardiomyopathy treatment.
    DOI:  https://doi.org/10.1155/2019/3206542
  90. Toxicology. 2019 Jul 29. pii: S0300-483X(19)30202-1. [Epub ahead of print] 152246
    Liu A, Xu X, Hou R, Badawy S, Tao Y, Chen D, Ihsan A, Wang X, Wu Q, Yuan Z.
      T-2 toxin is a secondary metabolite produced by Fusarium species and commonly contaminates food and animal feed. T-2 toxin can induce hepatotoxicity through apoptosis and oxidative stress; however, the underlying mechanism is not clear. Recent studies indicated that RASSF4, a member of the RASSF family, participates in cell apoptosis and some cancers due to its inactivation via DNA hypermethylation. However, its role in T-2 toxin-induced liver toxicity is poorly understood. Therefore, in this study, female Wistar rats were given a single dose of T-2 toxin at 2 mg/kg b.w. and were sacrificed at 1, 3 and 7 days post-exposure. A normal rat liver cell line (BRL) was exposed to different concentrations of T-2 toxin (10, 20, 40 nM) for 4, 8, 12 h, respectively. Histopathological analysis revealed with apoptosis in some liver cells and clear proliferation under T-2 toxin exposure. Expression analysis by immunohistochemical assays, quantitative real-time PCR (qPCR) and western blot demonstrated that T-2 toxin activated PI3K-Akt/Caspase/NF-κB signaling pathways. Additionally, DNA methylation assays revealed that the expression of RASSF4 was silenced by promoter hypermethylation after exposure to T-2 toxin for 1 and 3 days as compared to the control group. Moreover, joint treatment of 5-Aza-2'-deoxycytidine (DAC) (5 μM) and T-2 toxin (40 nM) increased expression of RASSF4 and PI3K-Akt/caspase/NF-κB signaling pathways-related genes, inducing cell apoptosis. These findings for the first time demonstrated that DNA methylation regulated the RASSF4 expression under T-2 toxin, along with the activation of its downstream pathways, resulting in apoptosis.
    Keywords:  DNA methylation; RASSF4; T-2 toxin; hepatotoxicity
    DOI:  https://doi.org/10.1016/j.tox.2019.152246
  91. Free Radic Res. 2019 Aug 02. 1-141
    Jiang Y, Xu , Yu L, Xu X, Feng C, Li J.
      Enteric glial cells (EGCs), one main cell population of the enteric nervous system (ENS), play a major role in regulating intestinal barrier function.Clostridium difficile toxin B(TcdB) is the major virulence factor produced by Clostridium difficile ( C. difficile ) and estimated to be toxic to EGCs by inducing cell death, cell cycle arrest and inflammatory cytokine production; however, the detailed mechanism for such effect is still unclear. In this study, we further evaluated the toxic effect of TcdB on EGCs and the involvement of NADPH oxidases in such process using the rat-transformed EGCs (CRL-2690). The results showed thatNOX4was activated by TcdB in EGCs and functioned as the major factor causing cytotoxicity and cell apoptosis. Mechanically, NOX4-generated H2O2 was the inducer of oxidative stress, Ca2+ homeostasis disorder and ER stress in EGCs upon TcdB treatment, and NOX4 inhibition protected EGCs against TcdB toxicity via attenuating these dysfunctions.These findings contribute to our understanding of the mechanism by which TcdB affects EGCs and suggest the potential value of NOX4 inhibition for treatment against C. difficile infection.
    Keywords:   toxin B; NOX4; endoplasmic reticulum stress; enteric glial cells; oxidative stress
    DOI:  https://doi.org/10.1080/10715762.2019.1649670
  92. Food Chem Toxicol. 2019 Jul 27. pii: S0278-6915(19)30509-5. [Epub ahead of print] 110719
    Abdelaziz AI, Mantawy EM, Gad AM, Fawzy HM, Azab SS.
      Methotrexate (MTX) is commonly used to treat several types of cancer and autoimmune diseases. However, there is increasing concern over its organs toxicities particularly liver toxicity. Liraglutide, a glucagon like peptide-1 agonist, possesses antioxidant and anti-inflammatory features. This study aimed to explore the potential protective effect of liraglutide pre-treatment in ameliorating MTX-induced hepatotoxicity and to further investigate the underlying mechanisms. Rats received 1.2 mg/kg liraglutide intraperitoneal twice daily for 7 days before MTX. Results revealed that liraglutide significantly decreased activities of liver enzymes and oxidative stress in hepatocytes. Furthermore, NF-kB expression and related inflammatory markers (TNF-α, COX-2 and IL-6) were reduced in the pre-treatment group of liraglutide. These data validate the advantageous effects of liraglutide in MTX hepatotoxic animals. In addition, liraglutide increased the expression of the antioxidant transcription factor nuclear factor-erythroid 2-related factor 2 (Nrf-2), along with the transcription of downstream phosphorylated cAMP response element-binding protein (pCREB) which increases the activity of Nrf-2. Additionally, caspase-3 expression/activity and BAX/Bcl-2 ratio were decreased following liraglutide pre-treatment. In conclusion, it was confirmed that liraglutide enhanced the antioxidant activity of liver cells by activating the Nrf-2 and pCREB signaling, thereby, reducing liver cell inflammation and apoptosis induced by MTX.
    Keywords:  HO-1; Heme oxygenase-1; Liraglutide; Methotrexate; Nrf 2; Nuclear factor erythroid 2-related factor2; P-CREB; Phosphorylated cAMP response element-binding protein
    DOI:  https://doi.org/10.1016/j.fct.2019.110719
  93. Front Pharmacol. 2019 ;10 746
    Li JJ, Yan YY, Sun HM, Liu Y, Su CY, Chen HB, Zhang JY.
      As a quinonemethide triterpenoid extracted from species of the Celastraceae and Hippocrateaceae, pristimerin has been shown potent anti-cancer effects. Specifically, it was found that pristimerin can affect many tumor-related processes, such as apoptosis, autophagy, migration and invasion, vasculogenesis, and drug resistance. Various molecular targets or signaling pathways are also involved, such as cyclins, reactive oxygen species (ROS), microRNA, nuclear factor kappa B (NF-κB), mitogen-activated protein kinase (MAPK), and PI3K/AKT/mammalian target of rapamycin (mTOR) pathways. In this review, we will focus on the research about pristimerin-induced anti-cancer activities to achieve a deeper understanding of the targets and mechanisms, which offer evidences suggesting that pristimerin can be a potent anti-cancer drug.
    Keywords:  anti-cancer; apoptosis; autophagy; mechanism; molecular target; pharmaceutical effect; pristimerin
    DOI:  https://doi.org/10.3389/fphar.2019.00746
  94. Environ Sci Technol. 2019 Jul 31.
    Zhou Q, Yue Z, Li Q, Zhou R, Liu L.
      PbSe nanoparticles (PbSe-NPs) attract ever-growing interests owing to great promise in various fields. However, potential toxic effects of PbSe-NPs on male reproductive systems have not been reported. This study aimed to determine whether early-life exposure to PbSe-NPs could affect male reproductive systems and other related health effects in rats. The male rats were intraperitoneally injected with 10 mg/kg/week PbSe-NPs for 60 days, followed by a series of reproductive-related analyses. We found that the nanoparticles could accumulate in testes in a size-dependent manner. Furthermore, accumulation of PbSe-NPs resulted in oxidative stress and disorder of normal serum sex hormones. Endoplasmic reticulum and mitochondria-mediated cell apoptosis were triggered via oxidative stress, as shown by upregulation of cytoplasmic Cyt-c, Bax, cleaved Caspase-3, GRP78, and Caspase-12. Notably, PbSe-NPs administration led to reduction in the quantity and quality of sperms, which caused a great fertility decrease. In contrast, released Pb2+ from PbSe-NPs did not result in any testes toxicity and fertility declines. These results demonstrate that PbSe-NPs could cause severe reproductive toxicity in a size-dependent manner and these toxic effects should contribute to PbSe-NPs themselves rather than released Pb2+. The application of PbSe-NPs might be a double-edged sword and corresponding measures should be taken before use.
    DOI:  https://doi.org/10.1021/acs.est.9b03581
  95. Sci Total Environ. 2019 Jul 18. pii: S0048-9697(19)33279-6. [Epub ahead of print]692 411-421
    Heredia-García G, Gómez-Oliván LM, Orozco-Hernández JM, Luja-Mondragón M, Islas-Flores H, SanJuan-Reyes N, Galar-Martínez M, García-Medina S, Dublán-García O.
      Sucralose (SUC) is an organochlorine that is used as a common sweetener in different dietary products around the world. Its extended use and production have led to this product is released into the environment in concentrations ranging from ng L-1 to μg L-1 in surface waters, groundwaters, wastewater treatment plants and ocean waters. A previous study carried out by our research team demonstrated that SUC is capable of inducing oxidative stress in Cyprinus carpio at environmentally-relevant concentrations. The aim of this study was to evaluate if SUC was capable of inducing alterations to DNA, apoptosis, and oxidative damage in the blood cells of C. carpio. Carps were exposed to two environmentally-relevant concentrations (0.05 and 155 μg L-1) of SUC, and the following biomarkers were determined: comet assay, micronucleus test (MN), caspase-3 activity, TUNEL assay, hydroperoxide content, lipid peroxidation level, protein carbonyl content and superoxide dismutase and catalase activities. Results obtained showed that SUC is capable of inducing DNA damage. A maximum increase of 35% and 23% were observed for c1 and c2, respectively in the comet assay; increases of 586% and 507.7% for c1 and c2, respectively, were found at 72 h through the MN test. The activity of caspase-3 showed a greater response for c1 and c2 at 96 h, with 271% and 493.5%, respectively. TUNEL assay also showed the highest response at 96 h, with 51.8 for c1 and 72.9 for c2; c1 y c2 were able to induce oxidative stress with the highest expression at 72 h. A correlation between DNA damage biomarkers, apoptosis and plasma levels of SUC in both concentrations were observed. With the data obtained, we can conclude that SUC, at environmentally-relevant concentrations, was capable of generating DNA alterations, apoptosis and oxidative stress in blood cells in common carp.
    Keywords:  Apoptosis; Common carp; DNA alterations; Oxidative stress; Sweeteners
    DOI:  https://doi.org/10.1016/j.scitotenv.2019.07.165
  96. Mitochondrion. 2019 Jul 26. pii: S1567-7249(19)30002-9. [Epub ahead of print]
    Miyauchi A, Kouga T, Jimbo EF, Matsuhashi T, Abe T, Yamagata T, Osaka H.
      Mitochondrial disease is a genetic disorder in which individuals suffer from energy insufficiency. The various clinical phenotypes of mitochondrial disease include Leigh syndrome (LS), myopathy encephalopathy lactic acidosis and stroke-like episodes (MELAS). Thus far, no curative treatment is available, and effective treatment options are eagerly awaited. We examined the cell protective effect of an existing commercially available chemical library on fibroblasts from four patients with LS and MELAS and identified apomorphine as a potential therapeutic drug for mitochondrial disease. We conducted a cell viability assay under oxidative stress induced by L-butionine (S, R)-sulfoximine (BSO), a glutathione synthesis inhibitor. Among the chemicals of library, 4 compounds (apomorphine, olanzapine, phenothiazine and ethopropazine) rescued cells from death induced by oxidative stress much more effectively than idebenone, which was used as a positive control. The EC50 value showed that apomorphine was the most effective compound. Apomorphine also significantly improved all of the assessed oxygen consumption rate values by the extracellular flux analyzer for fibroblasts from LS patients with complex I deficiency. In addition, the elevation of the Growth Differentiation Factor-15 (GDF-15), a biomarker of mitochondrial disease, was significantly reduced by apomorphine. Among 441 apomorphine-responsive genes identified by the microarray, apomorphine induced the expression of genes that inhibit the mammalian target of rapamycin (mTOR) activity and inflammatory responses, suggesting that apomorphine induced cell survival via a new potential pathway. In conclusion, apomorphine rescued fibroblasts from cell death under oxidative stress and improved the mitochondrial respiratory activity and appears to be potentially useful for treating mitochondrial disease.
    Keywords:  Apomorphine; Inflammation; Mammalian target of rapamycin (mTOR); Mitochondrial disease; Mitochondrial respiratory activity; Oxidative stress
    DOI:  https://doi.org/10.1016/j.mito.2019.07.006
  97. Arch Physiol Biochem. 2019 Aug 01. 1-7
    Ahangarpour A, Oroojan AA, Khorsandi L, Kouchak M, Badavi M.
      Context: The hyperglycemia (Hyper) induces oxidative stress in kidney tubular cells. Myricitrin (Myr) has an antioxidant effect along with low bioavailability. Objective: The present research investigated the effects of Myr and its solid lipid nanoparticles (SLN) on isolated proximal tubules exposed to the hyperglycemic condition. Materials and Methods: In this experimental study, the proximal tubules of mice were dissected by the microdissection method and the tubules were prepared for experimental or Real Time-PCR measurement. Results: The malondialdehyde level, transforming growth factor-β, nuclear factor kappa B and Bax genes expression increased in Hyper and decreased in Hyper + Myr and its SLN-treated groups compared to Hyper. Superoxide dismutase, total antioxidant capacity, the viability of proximal tubules and Bcl-2 gene expression decreased in untreated Hyper and increased in all treatment groups compared to Hyper. Conclusion: The hyperglycemia-induced oxidative disorder, inflammation and apoptosis in proximal tubules and that administrating Myr and its SLN improved them.
    Keywords:  Hyperglycemia; mouse; myricitrin; proximal tubule; solid lipid nanoparticle
    DOI:  https://doi.org/10.1080/13813455.2019.1647250
  98. Biochem Pharmacol. 2019 Jul 25. pii: S0006-2952(19)30278-3. [Epub ahead of print]
    Liu Q, Wen J, Zhu J, Zhang T, Deng Y, Jiang J.
      T-2 toxin is a secondary metabolite produced by the Fusarium genus and is highly toxic to both farmed animals and humans. In our previous study, we found that chicken cytochrome P450 1A5 (CYP1A5) can be significantly induced by T-2 toxin in chicken primary hepatocytes and catalyze T-2 toxin into a more toxic product, 3'-OH-T-2. Here, we showed that T-2 toxin also induced the expression of CYP1A5 in LMH cells at both the mRNA and protein levels, and this can be strongly inhibited by both resveratrol and siRNA targeting the aryl hydrocarbon receptor (AhR), indicating the involvement of AhR in T-2 toxin-induced transcriptional activation of CYP1A5. We further showed that T-2 toxin induced the expression of AhR and promoted the translocation of AhR into the nucleus as well as its binding to the proximal xenobiotic-responsive element (XRE) in the 5'-flanking region of CYP1A5, which mediates both the basal expression and the transcriptional activation of CYP1A5. Interestingly, CYP1A5 induction mediated by AhR enhances the cytotoxicity of T-2 toxin by reducing cell viability, activating oxidative stress and inducing DNA damage as well as apoptosis. Our findings provide novel insight into T-2 toxin-induced gene expression and cytotoxicity and may provide a novel target to reduce latent harm to chickens.
    Keywords:  AhR; CYP1A5; T-2 Toxin; cytotoxicity; transcriptional regulation
    DOI:  https://doi.org/10.1016/j.bcp.2019.07.023
  99. Am J Physiol Renal Physiol. 2019 Jul 31.
    Shaver CM, Paul MG, Putz ND, Landstreet SR, Kuck JL, Scarfe L, Skrypnyk NI, Yang H, Harrison FE, de Caestecker MP, Bastarache JA, Ware LB.
      Acute kidney injury is a common complication of severe sepsis and contributes to high mortality. The molecular mechanisms of acute kidney injury during sepsis are not fully understood. Because hemoproteins, including myoglobin and hemoglobin, mediate kidney injury during rhabdomyolysis, we hypothesized that cell-free hemoglobin would exacerbate acute kidney injury during sepsis. Sepsis was induced in mice by intraperitoneal injection of cecal slurry (CS). To mimic elevated levels of cell-free hemoglobin (CFH) observed during human sepsis, mice also received a retro-orbital injection of CFH or control. Four groups of mice were analyzed: sham, CFH alone, CS alone, and CS+CFH. The addition of CFH to CS reduced 48-hour survival compared to CS alone (67% vs 97%, p=0.001) and increased severity of illness. After 24 and 48 hrs, CS+CFH mice had reduced glomerular filtration rate (GFR) from baseline, while sham, CFH, and CS mice maintained baseline GFR. Biomarkers of acute kidney injury, NGAL and Kim-1, were markedly elevated in CS+CFH compared to CS (8-fold for NGAL, 2.4-fold for Kim-1, p<0.002 for each) after 48 hrs. Histologic examination showed a trend towards increased tubular injury in CS+CFH exposed kidneys compared to CS. However, there were similar levels of renal oxidative injury and apoptosis in CS+CFH compared to CS. Kidney levels of pro-inflammatory cytokines were similar between CS and CS+CFH. Human renal tubule cells (HK-2) exposed to CFH demonstrated increased cytotoxicity. Together, these results show that cell-free hemoglobin exacerbates acute kidney injury in a mouse model of experimental sepsis, potentially through increased renal tubular injury.
    Keywords:  acute kidney injury; cell-free hemoglobin; sepsis
    DOI:  https://doi.org/10.1152/ajprenal.00375.2018
  100. Diagn Microbiol Infect Dis. 2019 Jun 28. pii: S0732-8893(19)30644-3. [Epub ahead of print] 114860
    de Menezes RRPPB, Sampaio TL, Lima DB, Sousa PL, de Azevedo IEP, Magalhães EP, Tessarolo LD, Marinho MM, Dos Santos RP, Martins AMC.
      Chagas disease is caused by Trypanosoma cruzi and affects about 7 million people worldwide. Benznidazole and nifurtimox have low efficacy and high toxicity. The present study was designed to identify the trypanocidal effect of (-)-α-Bisabolol (BIS) and investigate its mechanism. Epimastigotes and trypomastigotes were cultured with BIS and the viable cells were counted. BIS antiamastigote effect was evaluated using infected LLC-MK2 cells. MTT assay was performed to evaluate BIS cytotoxicity. Growth recovery was assessed to evaluate BIS effect after short times of exposure. BIS mechanism was investigated through flow cytometry, with 7-AAD and Annexin V-PE. DCFH-DA, rhodamine 123 (Rho123) and acridine orange (AO). Finally, enzymatic and computational assays were performed to identify BIS interaction with T. cruzi GAPDH (tcGAPDH). BIS showed an inhibitory effect on epimastigotes after all tested periods, as well on trypomastigotes. It caused cytotoxicity on LLC-MK2 cells at higher concentrations, with selectivity index (SeI) = 26.5. After treatment, infected cells showed a decrease in infected cells, the number of amastigotes per infected cell and the survival index (SuI). Growth recovery demonstrated that BIS effect causes rapid death of T. cruzi. Flow cytometry showed that BIS biological effect is associated with apoptosis induction, increase in cytoplasmic ROS and mitochondrial transmembrane potential, while reservosome swelling was observed at a late stage. Also, BIS action mechanism may be associated to tcGAPDH inhibition. Altogether, the results demonstrate that BIS causes cell death in Trypanosoma cruzi Y strain forms, with the involvement of apoptosis and oxidative stress and enzymatic inhibition.
    Keywords:  Apoptosis; Bisabolol; Trypanosoma cruzi
    DOI:  https://doi.org/10.1016/j.diagmicrobio.2019.06.012
  101. Biosci Rep. 2019 Jul 30. pii: BSR20190548. [Epub ahead of print]
    Qin Y, Wang G, Peng Z.
      There is no effective treatment for septic acute kidney injury (AKI), which is considered a major public health concern in today's world. Here, we studied the functions of miR-191-5p in septic AKI. MiR-191-5p mimic or mimic control was injected into rats from caudal vein before Cecal ligation and puncture (CLP) surgery. Part of kidney tissues was stained by hematoxylin and eosin (H&E) for histological examination. The levels of serum cytokines were evaluated using enzyme-linked immunosorbent assay (ELISA). For cell transfection, renal cells were isolated from the kidneys of CLP rat model injected with mimic control and miR-191-5p mimic. With Targetscan prediction, serine/threonine-protein kinase OSR1 was identified as a target of miR-191-5p. Oxidative stress responsive 1 (OXSR1) overexpression vector was transfected into renal cells. Cell viability and apoptosis rate were determined by Cell Counting Kit-8 (CCK-8) and flow cytometry, respectively. We additionally measured the phosphorylation levels of p38 and p65. We found that the injection of miR-191-5p mimic could observably inhibit renal injury scores, and inhibit inflammatory cytokine productions and apoptotic protein levels in septic rats. After being transfected with OXSR1, the apoptosis rates and expressions of B-cell lymphoma-2 (Bcl-2), down-regulated Bax and Cleaved caspase-3 (C caspase-3) indicated overexpressed OXSR1 contributed to cell apoptosis. The up-regulated protein levels of p-p38 and p-p65 may suggest the involvement of p38 MAPK/NF-κB signaling pathway in the functions of OXSR1. Our results showed that the protective effects of miR-191-5p on kidney tissues of septic rats may rely on the repression of OXSR1.
    Keywords:  Apoptosis; Cecal ligation and puncture; Inflammatory cytokines; Septic acute kidney injury
    DOI:  https://doi.org/10.1042/BSR20190548
  102. Adv Exp Med Biol. 2019 ;1193 237-253
    Wu NN, Ren J.
      Aging is a complex irreversible biological process associated with increased prevalence of chronic disease and high healthcare burden. Several theories have been proposed for the biology of aging including free radical accumulation, DNA damage, apoptosis, telomere shortening, autophagy failure, and disturbed autonomic response. Aging is also closely associated with progressive deterioration of cardiovascular and neurological functions. Linkage, genome-wide association (GWAS), and next-generation sequencing analysis have confirmed a number of susceptibility loci for aging, in particular, Alzheimer's disease. Recent evidence from our group and others also revealed a tie between genetic mutation of mitochondrial aldehyde dehydrogenase (ALDH2) and life span as well as cardiovascular aging. ALDH2 represents the single most gene with the greatest number of human genetic polymorphism and is deemed an important enzyme for detoxification of reactive aldehydes. Here, we will briefly review the tie between ALDH2 and cardiovascular aging process. While recent work on ALDH2 research has broadened the pathogenic mechanisms of ALDH2 mutation or deficiency, therapeutic potential targeting ALDH2 in the elderly still remains debatable.
    Keywords:  ALDH2; Aging; Autophagy; Mitochondria; Oxidative stress
    DOI:  https://doi.org/10.1007/978-981-13-6260-6_15
  103. IUBMB Life. 2019 Jul 30.
    Ghanaatgar-Kasbi S, Amerizadeh F, Rahmani F, Hassanian SM, Khazaei M, Ferns GA, Avan A.
      Colorectal cancer (CRC) is among the leading causes of cancer-related mortality, despite extensive efforts in the identification of new treatment options. Hence, there is a need for the development of novel agents with therapeutic potential in treatment of CRC. Dorsomorphin has demonstrated antiproliferative activity against different malignancies. Here we have investigated the pharmaceutical potential of dorsomorphin in two-dimensional and three-dimensional cell-culture models of CRC. The antiproliferative, antimigratory, apoptotic activity and effect of this agent on cell cycle was evaluated by 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, wound healing assay, and flow cytometry, respectively, while the expression of genes involved in Wnt/Pi3K pathways was assessed at mRNA and/or proteins by reverse transcription polymerase chain reaction (RT-PCR) or western blot. Dorsomorphin inhibited CRC cell growth by modulating the cyclinD1, surviving and p-Akt. This agent was able to reduce the migratory behaviors of CRC cells, compared to control cells, through perturbation of E-cadherin. Also our data showed that dorsomorphin enhanced the percentage of the cells in sub-G1 and induced apoptosis in both late/early stages, as detected by annexin V. Also the regulatory effect of dorsomorphin on oxidant/antioxidant balance was assessed by cellular reactive oxygen species (ROS) generation. In particular, these data showed that dorsomorphin markedly increased the ROS production in CRC cells. Our finding demonstrated that dorsomorphin antagonizes cell growth and migration, through perturbation of Akt/mTOR/Wnt pathways in CRC, supporting further studies on the therapeutic potential of this novel anticancer agent in treatment of CRC.
    Keywords:  Wnt/PI3K/Akt -pathway; colorectal cancer; dorsomorphin
    DOI:  https://doi.org/10.1002/iub.2136
  104. Biol Pharm Bull. 2019 ;42(8): 1268-1274
    Wang X, Xiu Z, Du Y, Li Y, Yang J, Gao Y, Li F, Yin X, Shi H.
      Increasing evidence shows depression relevant to oxidative stress and inflammation. Anti-inflammatory strategies or antioxidants have led to the development of new antidepressants. Brazilin is a natural product from the Chinese traditional medicine Caesalpinia sappan L., exerting anti-inflammatory, antioxidant, anti-platelet concentration, and anti-cancer effects. While the antidepressant effect of brazilin is largely unknown. In present study, we investigated the effects of brazilin on H2O2-induced oxidative injury in PC12 cells and on depression- and anxiety-like behaviors of chronically mild stressed (CMS)-induced depression mice. It was found that brazilin pre-treatment (both 10 and 20 µM) significantly increased cell viability and decreased cell apoptosis in H2O2-treated PC12 cells. Furthermore, repetitive administration of brazilin to CMS-induced depression mice by intraperitoneal injection (10 mg/kg) made the mice significantly lose their latency of feeding in novelty-suppressed feeding test (NSF), have more the sucrose preference in sucrose preference test (SPT), and more time spent in the central zone without affecting their crossing activity in open field test (OFT). These results suggested that brazilin can play a role in antidepressant and anxiolytic-like behaviors for CMS-induced depression mice probably through inhibiting the oxidative stress. Therefore, brazilin is worth to be further explored for treating depressive and anxiety disorders.
    Keywords:  anxiety; brazilin; chronic mild stress; depression; oxidative stress
    DOI:  https://doi.org/10.1248/bpb.b18-00882