bims-aporos Biomed News
on Apoptosis and reactive oxygen species
Issue of 2019‒05‒19
fifty-seven papers selected by
Gavin McStay
Staffordshire University


  1. Oxid Med Cell Longev. 2019 ;2019 4824035
    Pu J, Zhu S, Zhou D, Zhao L, Yin M, Wang Z, Hong J.
      Background: The sedative anesthetic, propofol, is a cardioprotective agent for hyperglycemia-induced myocardial hypertrophy and dysfunction in rats. However, the specific protective mechanism has not been clarified.Methods and Results: In this experiment, we used H9c2 cells subjected to 22 mM glucose lasting for 72 hours as an in vitro model of cardiomyocyte injury by hyperglycemia and investigated the potential mechanism of propofol against hyperglycemic stress in cells. Propofol (5, 10, or 20 μM) was added to the cell cultures before and during the high glucose culture phases. Cell viability and levels of ROS were measured. The levels of proinflammatory cytokines were tested by ELISA. The levels of SIRT3, SOD2, PHD2, HIF-1α, Bcl-2, P53, and cleaved caspase-3 proteins were detected by western blotting. Our data showed that propofol attenuated high glucose-induced cell apoptosis accompanied by a decrease in the level of reactive oxygen species (ROS) and proinflammatory cytokines. Meanwhile, propofol decreased the apoptosis of H9c2 cells via increasing the expression of Bcl-2, SIRT3, SOD2, and PHD2 proteins and decreasing the expression of cleaved caspase-3, P53, and HIF-1α. Real-time PCR analysis showed that propofol did not significantly change the HIF-1α but increase PHD2 at mRNA level. HIF-1α silence significantly decreased apoptosis and inflammation in H9c2 cell during high glucose stress. Pretreatment of IOX2 (the inhibitor of PHD2) inhibited cell viability until the concentration reached 200 μM during high glucose stress. However, 50 μM TYP (the inhibitor of SIRT3) significantly inhibited cell viability during high glucose stress. Delayed IOX2 treatment for 6 hours significantly inhibited cell viability during high glucose stress.
    Conclusions: Propofol might alleviate cell apoptosis via SIRT3-HIF-1α axis during high glucose stress.
    DOI:  https://doi.org/10.1155/2019/4824035
  2. J Biochem Mol Toxicol. 2019 May 13. e22326
    Yildizbayrak N, Erkan M.
      The aim of this study was to investigate the possible therapeutic effects of curcumin (CUR), against acrylamide (AA)-induced toxic effects on Leydig cells. The AA and CUR-treated cells were evaluated for cell viability, lipid peroxidation, reactive oxygen species (hydroxyl radical and hydrogen peroxide), antioxidant levels (glutathione peroxidase, glutathione-S-transferase, and catalase), apoptosis/necrosis rates and phosphorylation status of mitogen-activated protein kinases (MAPKs). Leydig cells were exposed to four concentrations of AA (1, 10, 100, 1000 µM) in the presence and absence of CUR (2.5 µM) for 24 hours. According to the present result, AA concentration-dependently, increased the oxidative stress parameters and suppressed the antioxidant enzyme levels, meanwhile induced apoptosis and activated the phosphorylation of extracellular signal-regulated kinase, p38, and c-Jun NH 2 -terminal kinase. Moreover, CUR ameliorated the detrimental effects of AA. Thus, AA-induced apoptosis through activation of the MAPK signaling pathway and CUR has a protective effect against AA-induced damage in Leydig cells.
    Keywords:  Leydig cell; acrylamide; apoptosis; curcumin; mitogen-activated protein kinases
    DOI:  https://doi.org/10.1002/jbt.22326
  3. Appl Biochem Biotechnol. 2019 May 16.
    Xu J, Cai X, Teng S, Lu J, Zhou Y, Wang X, Meng Z.
      Based on the various pharmacological activities of tamarixetin, the present study investigated the cytotoxicity property of tamarixetin in human liver cancer cells including PLC/PRF/5 and HepG2 cells, and their xenografted tumor nude mice. In cells, tamarixetin incubation resulted in the suppression on cell viability; enhanced cell apoptosis rate, LDH release, caspase-3 activation, and reactive oxygen species accumulation; and decreased mitochondrial membrane potential in a dose-dependent manner. Tamarixetin inhibited the growth of PLC/PRF/5- and HepG2-xenografted tumors in BALB/c nude mice after 14-day administration without influencing their bodyweights and organ functions including liver and spleen. Tamarixetin enhanced the expression levels of pro-apoptotic proteins including Bax and cleaved caspase-3 and inhibited the expression levels of anti-apoptotic proteins including Bcl-2 and Bcl-xL in liver cancer cells and their xenografted tumor tissues. Furthermore, tamarixetin significantly suppressed the phosphorylation of ERKs and AKT in both PLC/PRF/5 and HepG2 cells, and tumor tissues. All present data suggest that tamarixetin displays pro-apoptotic properties in liver cancer cells related to the mitochondria apoptotic pathway via regulating the ERKs and AKT signaling.
    Keywords:  Apoptosis; ERKs and AKT; Liver cancer; Mitochondria; Tamarixetin
    DOI:  https://doi.org/10.1007/s12010-019-03033-x
  4. Dalton Trans. 2019 May 13.
    Yu T, Zhen M, Li J, Zhou Y, Ma H, Jia W, Wang C.
      Metallofullerene Gd@C82 derivatives have been found to exhibit excellent performance in the treatment of chronic diseases and in tumor therapy due to their excellent anti-oxidation ability. Thus, there is a great need to clarify the specific protection mechanism of Gd@C82 derivatives against oxidative stress at the cellular and molecular level. Herein, we optimize the preparation of amino acid derivatives of Gd@C82 (GF-Ala) and investigated the potential mechanism of anti-oxidative stress induced by H2O2 in L02 human hepatic cells. GF-Ala with excellent biocompatibility and high production yield was obtained by adjusting the size of solid Gd@C82 as a starting material. Pretreatment with GF-Ala significantly improved the cell viability of oxidatively damaged cells. Furthermore, we found that GF-Ala prominently reduced intracellular reactive oxygen species (ROS) production, stabilized mitochondrial membrane potential (MMP) and inhibited cell apoptosis. Moreover, GF-Ala up-regulated the expression of Bcl-2 and down-regulated the expression of Bax, which further prevented the activation of pro-caspase 3 and PARP. Thus, we showed that GF-Ala protected the cells from oxidative stress by blocking the mitochondria-mediated apoptosis pathway.
    DOI:  https://doi.org/10.1039/c9dt00800d
  5. Sci Total Environ. 2019 May 05. pii: S0048-9697(19)32056-X. [Epub ahead of print]679 365-377
    Khan I, Bahuguna A, Krishnan M, Shukla S, Lee H, Min SH, Choi DK, Cho Y, Bajpai VK, Huh YS, Kang SC.
      Human health and environment have been continuously getting exposure to toxic chemicals including nanomaterial; therefore, nontoxicity has recently attracted huge amount of attention. In this study, RU-AgNPs were synthesized by a green synthesis procedure and evaluated for their toxicity in human umbilical vein endothelial cells (HUVECs) as well as on zebrafish embryos via apoptotic pathway. The synthesized RU-AgNPs were average in size (20-25 nm) with a negative surface charge of -13.43 mV. As a result, RU-AgNPs potentiated the formation of reactive oxygen species (ROS) in HUVECs as confirmed by the results of immunoblotting analysis using apoptotic markers, such as Bax, Bcl2, and cytochrome C. Moreover, the induction of apoptosis in HUVECs was also authenticated in a dose-dependent manner after the treatment with RU-AgNPs by the Incucyte analysis. In vivo trials conducted on zebrafish visualized the mortality, malformation, and imbalanced in the heart rate, and cell death of the whole embryo, including severe morphological changes in the yolk sac and the tail of zebrafish. Furthermore, the results of western blot analysis demonstrated the increasing intensity of apoptotic biomarkers such as Bax, Bcl2, and Cyto C, including enhanced production of ROS, validating the cell death in zebrafish larvae. In addition, chemically functionalized silver nanoparticles found to be more cytotoxic than biogenic functionalized silver nanoparticles. Above-mentioned findings clearly demonstrate that Ru-AgNPs cause the toxicity via ROS-induced apoptotic pathway. Therefore, it is necessary to decide RU-AgNPs toxicity levels before being used in any biomedical application.
    Keywords:  Apoptosis; HUVECs; ROS; RU-AgNPs; Zebrafish
    DOI:  https://doi.org/10.1016/j.scitotenv.2019.05.045
  6. Toxins (Basel). 2019 May 11. pii: E265. [Epub ahead of print]11(5):
    Habrowska-Górczyńska DE, Kowalska K, Urbanek KA, Domińska K, Sakowicz A, Piastowska-Ciesielska AW.
      Deoxynivalenol (DON), known as vomitoxin, a type B trichothecene, is produced by Fusarium. DON frequently contaminates cereal grains such as wheat, maize, oats, barley, rye, and rice. At the molecular level, it induces ribosomal stress, inflammation and apoptosis in eukaryotic cells. Our findings indicate that DON modulates the viability of prostate cancer (PCa) cells and that the response to a single high dose of DON is dependent on the androgen-sensitivity of cells. DON appears to increase reactive oxygen species (ROS) production in cells, induces DNA damage, and triggers apoptosis. The effects of DON application in PCa cells are influenced by the mitogen-activated protein kinase (MAPK) and NFΚB- HIF-1α signaling pathways. Our results indicate that p53 is a crucial factor in DON-associated apoptosis in PCa cells. Taken together, our findings show that a single exposure to high concentrations of DON (2-5 µM) modulates the progression of PCa.
    Keywords:  apoptosis; deoxynivalenol; mycotoxin; oxidative stress; prostate cancer
    DOI:  https://doi.org/10.3390/toxins11050265
  7. Cells. 2019 May 10. pii: E444. [Epub ahead of print]8(5):
    Gurunathan S, Jeyaraj M, Kang MH, Kim JH.
      Generally, platinum nanoparticles (PtNPs) are considered non-toxic; however, toxicity depends on the size, dose, and physico-chemical properties of materials. Owing to unique physico-chemical properties, PtNPs have emerged as a material of interest for several biomedical applications, particularly therapeutics. The adverse effect of PtNPs on the human monocytic cell line (THP-1) is not well-established and remains elusive. Exposure to PtNPs may trigger oxidative stress and eventually lead to inflammation. To further understand the toxicological properties of PtNPs, we studied the effect of biologically synthesized ultra-small PtNPs on cytotoxicity, genotoxicity, and proinflammatory responses in the human monocytic cell line (THP-1). Our observations clearly indicated that PtNPs induce cytotoxicity in a dose-dependent manner by reducing cell viability and proliferation. The cytotoxicity of THP-1 cells correlated with an increase in the leakage of lactate dehydrogenase, generation of reactive oxygen species, and production of malondialdehyde, nitric oxide, and carbonylated proteins. The involvement of mitochondria in cytotoxicity and genotoxicity was confirmed by loss of mitochondrial membrane potential, lower ATP level, and upregulation of proapoptotic and downregulation of antiapoptotic genes. Decreases in the levels of antioxidants such as reduced glutathione (GSH), oxidized glutathione (GSH: GSSG), glutathione peroxidase (GPx), superoxide dismutase (SOD), catalase (CAT), and thioredoxin (TRX) were indicative of oxidative stress. Apoptosis was confirmed with the significant upregulation of key apoptosis-regulating genes. Oxidative DNA damage was confirmed by the increase in the levels of 8-oxodG and 8-oxoG and upregulation of DNA damage and repair genes. Finally, the proinflammatory responses to PtNPs was determined by assessing the levels of multiple cytokines such as interleukin-1β (IL-1β), IL-6, IL-8, tumor necrosis factor-α (TNF-α), granulocyte-macrophage colony-stimulating factor (GM-CSF), and monocyte chemoattractant protein 1 (MCP-1). All the cytokines were significantly upregulated in a dose-dependent manner. Collectively, these observations suggest that THP-1 cells were vulnerable to biologically synthesized ultra-small PtNPs.
    Keywords:  DNA damage; apoptosis; cytokines; genotoxicity; oxidative stress; platinum nanoparticles; proinflammatory response
    DOI:  https://doi.org/10.3390/cells8050444
  8. Biochem Biophys Res Commun. 2019 May 14. pii: S0006-291X(19)30854-X. [Epub ahead of print]
    Zheng H, Zhang M, Luo H, Li H.
      Ultraviolet B (UVB) irradiation increases the risk of various skin disorders, resulting in apoptosis, autophagy and oxidative stress and thereby promoting the risk of skin photoaging and carcinogenesis. The use of photochemoprotectors including natural products with antioxidant properties represents an effective strategy for preventing UVB-induced skin injury. Isoorientin (Iso), as a flavonoid compound, could be extracted from several plant species and possesses multiple biological activities. However, its role in regulating UVB-induced skin damage is little to be reported. In the study, we found that Iso treatment could protect human dermal fibroblasts (HDFs) against the effects of UVB irradiation by improving cell viability, suppressing MMP1 and MMP3 expression, inhibiting oxidative stress and inducing autophagy. In addition, Iso reduced UVB-triggered apoptosis, as evidenced by the decreased Caspase-3 activity in vitro. Furthermore, Iso was functioned as reactive oxygen species (ROS) scavenger that markedly hindered c-Jun N-terminal kinases (JNK) signaling activation in UVB-treated HFDs. Importantly, promoting JNK activity restored matrix metalloproteinase (MMP)-1/3 expression in Iso-incubated cells with UVB stimulation. Meanwhile, UVB exposure to the skin of mice and subsequent topical application of Iso delayed the progression of skin damage, resulting in autophagy and blocking the JNK activation and ROS production. In conclusion, these results indicated the photoprotective role of Iso and demonstrated that Iso could also be potentially used as an agent against UVB-stimulated skin damage.
    Keywords:  Autophagy; Isoorientin; JNK; ROS; UVB irradiation
    DOI:  https://doi.org/10.1016/j.bbrc.2019.04.195
  9. Life Sci. 2019 May 08. pii: S0024-3205(19)30347-9. [Epub ahead of print]
    Mortezaee K, Najafi M, Farhood B, Ahmadi A, Potes Y, Shabeeb D, Musa AE.
      Radio- and chemotherapy are the most common cancer treatment modalities. They cause acute and late side effects on normal tissues, which is a burden for delivery of a high dose of radiation or drugs on tumor cells. In addition, tumor cells achieve adaptive responses to subsequent doses of radiation and chemotherapy, leading to tumor resistance and accelerated repopulation. Resistance to radiotherapy and chemotherapy can occur following adaptive responses, which itself is due to the release of large numbers of inter- and intracellular mediators by immune cells as well as other tumor microenvironment (TME) cells. Melatonin is a potent natural antioxidant and anti-inflammatory agent that protects against toxic side effects of radiation and chemotherapy. Furthermore, in some cancer cells, melatonin aids sensitizing cancer cells to therapy. Apoptosis is one of the main mechanisms of cell death following exposure to radiation and chemotherapy. Evidences have shown a direct relation between apoptosis induction in tumor cells with increased tumor delay regression and survival. Melatonin through modulation of several apoptosis mediators such as mitochondria, Bax, Bcl-2, endogenous ROS, and apoptosis receptors facilitate apoptosis. The current review aims to explain mechanisms of apoptosis induction following exposure to radiation and chemotherapy drugs. We also reviewed the modulatory effect of melatonin on apoptosis signaling pathways.
    Keywords:  Apoptosis; COX-2; Chemotherapy; Hypoxia; MAPKs; Melatonin; Mitochondria; NF-κB; Neoplasm; Radiation; Radiotherapy; Tumor Microenvironment (TME)
    DOI:  https://doi.org/10.1016/j.lfs.2019.05.009
  10. J Clin Med. 2019 May 14. pii: E680. [Epub ahead of print]8(5):
    Khan A, Ikram M, Muhammad T, Park J, Kim MO.
      Cadmium (Cd), a nonbiodegradable heavy metal and one of the most neurotoxic environmental and industrial pollutants, promotes disturbances in major organs and tissues following both acute and chronic exposure. In this study, we assessed the neuroprotective potential of caffeine (30 mg/kg) against Cd (5 mg/kg)-induced oxidative stress-mediated neuroinflammation, neuronal apoptosis, and cognitive deficits in male C57BL/6N mice in vivo and in HT-22 and BV-2 cell lines in vitro. Interestingly, our findings indicate that caffeine markedly reduced reactive oxygen species (ROS) and lipid peroxidation (LPO) levels and enhanced the expression of nuclear factor-2 erythroid-2 (Nrf-2) and hemeoxygenase-1 (HO-1), which act as endogenous antioxidant regulators. Also, 8-dihydro-8-oxoguanine (8-OXO-G) expression was considerably reduced in the caffeine-treated group as compared to the Cd-treated group. Similarly, caffeine ameliorated Cd-mediated glial activation by reducing the expression of glial fibrillary acidic protein (GFAP), ionized calcium-binding adapter molecule 1 (Iba-1), and other inflammatory mediators in the cortical and hippocampal regions of the mouse brain. Moreover, caffeine markedly attenuated Cd-induced neuronal loss, synaptic dysfunction, and learning and cognitive deficits. Of note, nuclear factor-2 erythroid-2 (Nrf-2) gene silencing and nuclear factor-κB (NF-κB) inhibition studies revealed that caffeine exerted neuroprotection via regulation of Nrf-2- and NF-κB-dependent mechanisms in the HT-22 and BV-2 cell lines, respectively. On the whole, these findings reveal that caffeine rescues Cd-induced oxidative stress-mediated neuroinflammation, neurodegeneration, and memory impairment. The present study suggests that caffeine might be a potential antioxidant and neuroprotective agent against Cd-induced neurodegeneration.
    Keywords:  Nrf-2/HO-1; cadmium; caffeine; memory impairments; neurodegeneration; p-NF-κB; reactive oxygen species (ROS)
    DOI:  https://doi.org/10.3390/jcm8050680
  11. Environ Pollut. 2019 May 06. pii: S0269-7491(19)30837-1. [Epub ahead of print]251 372-379
    Yang X, Zhang X, Zhang J, Ji Q, Huang W, Zhang X, Li Y.
      T-2 toxin is an unavoidable contaminant in human food, animal feeds, and agricultural products. T-2 toxin has been found to impair male reproductive function. But, few data is available that reveals the reproductive toxicity mechanism. In the study, male Kunming mice were orally administrated with T-2 toxin at the doses of 0, 0.5, 1 or 2 mg/kg body weight for 28 days. The body and reproductive organs weight, the concentration, malformation rate and ultrastructure of sperm in cauda epididymis were detected. Oxidative stress biomarkers and apoptosis were also measured in testes. Histological change of testes was performed by H&E and TUNEL staining. T-2 toxin down-regulated body and reproductive organs (testis, epididymis and seminal vesicle) weight, sperm concentration, increased sperm malformation rate and damaged the ultrastructure of sperm and structure of testes. T-2 toxin treatment increased the reactive oxygen species (ROS) and malondialdehyde content, while, decreased the total anti-oxidation capacity (T-AOC) and the superoxide dismutase activity in testes. T-2 toxin exposure increased the TUNEL-positive germ cells, the activities and mRNA expressions of caspase-3, caspase-8 and caspase-9, the mRNA expression of Bax, and inhibited the Bcl-2 mRNA expression. Furthermore, the expressions of caspase-3, caspase-8 caspase-9 and Bax were positively correlated with ROS level, but negatively correlated with T-AOC in testis. In summary, T-2 toxin caused spermatogenesis disorder associated with the germ cell apoptosis medicated by oxidative stress, impairing the male reproductive function.
    Keywords:  Apoptosis; Oxidative stress; Spermatogenesis; T-2 toxin; Testis
    DOI:  https://doi.org/10.1016/j.envpol.2019.05.023
  12. Toxins (Basel). 2019 May 15. pii: E273. [Epub ahead of print]11(5):
    Radovanović V, Vlainić J, Hanžić N, Ukić P, Oršolić N, Baranović G, Jazvinšćak Jembrek M.
      Elevated amounts of copper are considered to be contributing factor in the progression of neurodegenerative diseases as they promote oxidative stress conditions. The aim of our study was to examine the effects of ethanolic extract of propolis (EEP) against copper-induced neuronal damage. In cultured P19 neuronal cells, EEP exacerbated copper-provoked neuronal cell death by increasing the generation of reactive oxygen species (ROS) and through the activation of caspase-3/7 activity. EEP augmented copper-induced up-regulation of p53 and Bax mRNA expressions. Neurotoxic effects of EEP were accompanied by a strong induction of glyceraldehyde 3-phosphate dehydrogenase (GAPDH) expression and decrease in the expression of c-fos mRNA. SB203580, an inhibitor of p38 mitogen-activated protein kinase (MAPK) prevented detrimental effects of EEP, whereas SP600125, an inhibitor of c-Jun N-terminal kinase (JNK), exacerbated EEP-induced neuronal cell death. Quercetin, a polyphenolic nutraceutical, which is usually present in propolis, was also able to exacerbate copper-induced neuronal death. Our data indicates a pro-oxidative and apoptotic mode of EEP action in the presence of excess copper, wherein ROS/p53/p38 interactions play an important role in death cascades. Our study also pointed out that detailed pharmacological and toxicological studies must be carried out for propolis and other dietary supplements in order to fully recognize the potential adverse effects in specific conditions.
    Keywords:  P19 neurons; copper toxicity; ethanolic extract of propolis; p38 and JNK signaling; p53
    DOI:  https://doi.org/10.3390/toxins11050273
  13. Biochem Pharmacol. 2019 May 09. pii: S0006-2952(19)30177-7. [Epub ahead of print]
    Ye J, Ma J, Liu C, Huang J, Wang L, Zhong X.
      Esophageal squamous cell carcinoma (ESCC) is one of the most common and aggressive cancers worldwide, especially in China, with poor prognosis due to the lack of effective therapeutic strategies. Here, the anticancer effect and pharmacological mechanism of a newly synthesized Fe(II) phenanthroline complex was studied in ESCC. Our data showed that transferrin receptor 1 (TFR1) was specifically overexpressed in ESCC tissues compared to its expression in normal esophageal tissues, a finding further supported by public datasets. The newly synthesized Fe(II) complex was selectively transported into ESCC cells overexpressing TFR1 through TFR1-mediated endocytosis and exhibited anticancer activity in a dose-dependent manner. The mechanistic study elucidated that the Fe(II) complex caused cell cycle arrest at the G0/G1 phase by blocking the CDK4/6-cyclin D1 complex and induced mitochondria-mediated apoptosis. Furthermore, exposure to the Fe(II) complex led to excessive reactive oxygen species (ROS) accumulation by thioredoxin reductase (TrxR) inhibition and DNA double-strand breaks (DSBs), which in turn sequentially activated ATM, CHK1/2 and p53. Moreover, combination treatment with cisplatin and the Fe(II) complex exhibited a synergistic effect in ESCC cells. Taken together, our results initially suggest the potential application of the Fe(II) complex in ESCC chemotherapy, especially for patients with TFR1 overexpression.
    Keywords:  DNA damage; Iron(II) phenanthroline complex; esophageal squamous cell carcinoma; reactive oxygen species; thioredoxin reductase; transferrin receptor 1
    DOI:  https://doi.org/10.1016/j.bcp.2019.05.013
  14. Oxid Med Cell Longev. 2019 ;2019 6051262
    Li T, Xian WJ, Gao Y, Jiang S, Yu QH, Zheng QC, Zhang Y.
      Hypoxia-inducible gene domain family member 1A (Higd1a) has recently been reported to protect cells from hypoxia by helping to maintain normal mitochondrial function. The potential induction of Higd1a under high-fat exposure and whether it could protect cells from oxidative stress attracted our attention. Initially, 0.4 mM oleic acid and 0.2 mM palmitate were added to the growth media of HepG2 and LO2 cells for 72 hours. We discovered increased Higd1a expression, and knocking down Higd1a impaired mitochondrial transmembrane potential and induced cell apoptosis. We then identified that elevated reactive oxygen species (ROS) is responsible for increased Higd1a expression. Furthermore, we found that ROS promoted Higd1a expression by upregulating HIF-1a and PGC-1a expressions, and these two proteins could exert synergistic effects in inducing Higd1a expression. Taken together, these data suggest that Higd1a plays positive roles in protecting cells from oxidative stress, and ROS could induce Higd1a expression by upregulating PGC-1a and HIF-1a expressions.
    DOI:  https://doi.org/10.1155/2019/6051262
  15. Nutrients. 2019 May 11. pii: E1061. [Epub ahead of print]11(5):
    Kim JL, Park SH, Jeong S, Kim BR, Na YJ, Jo MJ, Jeong YA, Yun HK, Kim DY, Kim BG, You S, Oh SC, Lee DH.
      Natural products have shown great promise in sensitizing cells to TNF-related apoptosis-inducing ligand (TRAIL) therapy. Sea cucumber (SC) extracts possess antitumor activity, and hence their potential to sensitize colorectal cancer (CRC) cells to TRAIL therapy was evaluated. This study used Western blotting to evaluate the combination effects of SC and TRAIL in CRC, and determined the molecular mechanism underlying these effects. SC fractions and TRAIL alone did not affect apoptosis; however, combined treatment dramatically induced the apoptosis of CRC cells, but not of normal colon cells. Combined treatment induced the expression of apoptotic proteins (poly (ADP-ribose) polymerase (PARP), caspase 3, and 8), and this effect was markedly inhibited by the ubiquitination of X-linked inhibitor of apoptosis protein (XIAP). SC did not affect the mRNA levels, but it increased proteasomal degradation and ubiquitination of the XIAP protein. Furthermore, SC induced reactive oxygen species (ROS) production, thereby activating c-Jun N-terminal kinase (JNK) and endoplasmic reticulum (ER) stress-related apoptotic pathways in CRC. Altogether, our results demonstrate that the SC F2 fraction may sensitize CRC cells to TRAIL-induced apoptosis through XIAP ubiquitination and ER stress.
    Keywords:  X-linked inhibitor of apoptosis protein (XIAP); c-Jun N-terminal kinase (JNK); endoplasmic reticulum (ER) stress; sea cucumber (SC)
    DOI:  https://doi.org/10.3390/nu11051061
  16. Ecotoxicol Environ Saf. 2019 May 14. pii: S0147-6513(19)30541-X. [Epub ahead of print]180 280-287
    Liu J, Zhang J, Ren L, Wei J, Zhu Y, Duan J, Jing L, Sun Z, Zhou X.
      Fine particulate matters (PM2.5) have been associated with male reproductive toxicity because it can penetrate into the lung's gas-exchange region, and spread to the whole body via circulatory system. Previous studies have shown that PM2.5 could induce DNA damage and apoptosis by reactive oxygen species (ROS). The aim of the present study is to determine the exact mechanism and role of apoptosis induced by PM2.5 in spermatocyte cells. Male Sprague-Dawley (SD) rats were treated with normal saline (control group) or PM2.5 with the doses of 1.8, 5.4 and 16.2 mg/kg bw. via intratracheal instillation every 3 days for 30 days. Mouse spermatocyte-derived cells (GC-2spd cells) were treated with various concentrations (0, 50, 100, 200 μg/mL) of PM2.5 for 24 h. The results showed that exposure to PM2.5 resulted in injury of testicular tissue and impaired mitochondria integrity in GC-2spd cells. Moreover, PM2.5 induced DNA damage and apoptosis in GC-2spad cells via ROS generation, and the ATM/P53/CDK2 and mitochondria apoptosis pathway autophagy signal pathway were activated. N-Acetyl-L-cysteine (NAC), a well-known antioxidant, ameliorated DNA damage, and inhibited apoptosis. These findings demonstrated PM2.5 might induce apoptosis via the mitochondrial apoptosis pathway through causing DNA damage resulting from oxidative stress, and finally caused spermatogenesis disorder.
    Keywords:  ATM/P53/CDK2 pathway; Fine particulate matters; GC-2spd cells; Mitochondrial apoptosis pathway; Oxidative stress
    DOI:  https://doi.org/10.1016/j.ecoenv.2019.05.013
  17. Exp Ther Med. 2019 Jun;17(6): 4409-4416
    Wang J, Guo HM.
      Diabetic nephropathy (DN) has become the major cause of end-stage renal disease increasing the mortality risk of diabetes. Research has demonstrated that the oxidative damage and apoptosis of renal tubular cells is present during DN. Astragaloside IV (AS-IV) has been widely used for the treatment of many diseases, however, the role and mechanism by which AS-IV may ameliorate high glucose-induced apoptosis and oxidative stress of the human proximal tubular cell line HK-2 remains largely unknown. The present study investigated the effect of AS-IV on high glucose-induced apoptosis and oxidative stress in HK-2 cells. Cell viability, apoptosis and protein expression were detected by Trypan blue staining, Cell Counting Kit-8 assay, terminal deoxynucleotidyl transferase 2'-deoxyuridine-5'-triphosphate nick-end labelling, flow cytometry and western blot analyses. In addition, enzymatic activities, including superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), catalase (CAT) and lipid peroxide (LPO), were measured with the corresponding detection kits. DCFH-DA assay and flow cytometry were performed to detect the production of reactive oxygen species (ROS). Western blot analysis and reverse transcription-quantitative polymerase chain reaction were conducted to evaluate protein and mRNA expressions of the nuclear factor erythroid 2 like 2 (Nrf2)/antioxidant response element (ARE) signaling pathway. The results demonstrated that AS-IV significantly enhanced HK-2 cell viability induced by high glucose in a dose-dependent manner. In addition, AS-IV notably inhibited HK-2 cell apoptosis stimulated by high glucose, which may be associated with inhibition of BCL2 associated X protein, Cleaved-caspase-3 and Cleaved-caspase-9, expression and the promotion of Bcl-2. AS-IV significantly increased the activities of antioxidant enzymes SOD, GSH-Px and CAT, and decreased the high-glucose-induced ROS production in HK-2 cells, in a dose-dependent manner. Finally, it was determined that AS-IV regulated the Nrf2/ARE signaling pathway and inhibited the expression of liver-type fatty acid binding protein. In conclusion, these findings may provide evidence that AS-IV has a potential role for the treatment of DN.
    Keywords:  HK-2 cells; apoptosis; astragaloside IV; high glucose; nuclear factor erythroid 2 like 2/antioxidant response element signaling pathway; oxidative stress
    DOI:  https://doi.org/10.3892/etm.2019.7495
  18. Eur Rev Med Pharmacol Sci. 2019 Apr;pii: 17675. [Epub ahead of print]23(8): 3173-3182
    Zhao J, Sun H, Zhang JM, Wang M, Du XJ, Zhang JL.
      OBJECTIVE: Although the potential involvements of INK4 locus reported in glaucoma, the effects of long non-coding RNA (lncRNA) antisense noncoding RNA in the INK4 locus (ANRIL) on trabecular meshwork (TM) cells remain unclear. In this study, we aimed to explore the effects of lncRNA ANRIL on the oxidative injury of human TM cells as well as the underlying mechanisms.MATERIALS AND METHODS: Oxidative injury of human TM cells was induced by H2O2 stimulation. Cell viability, apoptotic cells, expression of proteins related to apoptosis, and reactive oxygen species (ROS) level were testified by CCK-8 assay, flow cytometry assay, Western blot analysis, and DCFH-DA staining, respectively. LncRNA ANRIL was overexpressed, and its effects on H2O2-injured TM cells were analyzed. Afterward, miR-7 expression in lncRNA ANRIL overexpressing-cells was determined by RT-qPCR. Moreover, it was verified whether lncRNA ANRIL affected H2O2-treated TM cells via miR-7, followed by the measurements of the involved signaling pathways.
    RESULTS: H2O2-induced decrease of cell viability and the increases in apoptosis and ROS generation were significantly attenuated by lncRNA ANRIL overexpression. miR-7 expression was down-regulated by lncRNA ANRIL, and miR-7 overexpression abrogated the effects of lncRNA ANRIL on H2O2-treated TM cells. Phosphorylation levels of the key kinases in the mTOR and MEK/ERK pathways were enhanced by lncRNA ANRIL via down-regulating miR-7 in H2O2-treated TM cells.
    CONCLUSIONS: LncRNA ANRIL attenuated oxidative injury of human TM cells and activated the mTOR and MEK/ERK pathways, possibly through down-regulating miR-7.
    DOI:  https://doi.org/10.26355/eurrev_201904_17675
  19. J Cell Biochem. 2019 May 17.
    Eraslan E, Tanyeli A, Polat E, Yetim Z.
      Renal ischemia/reperfusion (I/R) injury resulting in acute renal failure, is a major clinical problem due to its high mortality rate. Renal I/R increases the reactive oxygen species, secretion of inflammatory cytokines, chemokines and other factors. This suggests that initiating the apoptosis process in the presence of oxidative stress may play a role in life-threatening conditions, such as ischemia. Ischemia reperfusion-induced renal damage can result in renal failure and death. Although many treatment procedures have been carried out to reduce or destroy renal I/R damage in experimental models, so far, a routine method of treatment has not yet been found. For this reason, the current study was planned to investigate the possible protective effects of evodiamine on tissue damage caused by ischemia-reperfusion in kidney tissue in rats and an experimental renal I/R model was used for this purpose. Four groups were formed in the study: the control, sham control, ischemia reperfusion (I/R), and evodiamine (10 mg/kg) + I/R groups. The effects of evodiamine against kidney I/R injury were investigated. TAS (total oxidant status), TOS (total oxidant status), interleukin-1β (IL-1β), IL-6, IL-10 and tumor necrosis factor-α levels were determined by enzyme-linked immunosorbent assay. The oxidative stress index was calculated from TAS and TOS levels. In addition, the renal ischemia reperfusion injury was examined histopathologically. The IL-10 and TAS levels in the I/R group decreased when compared with the control and Sham groups, while these levels increased in the evodiamine group. Histopathologic examination revealed that caspase 3 and nuclear factor-κB levels decreased in the evodiamine group compared with the I/R group. The application of evodiamine significantly reduced ischemia reperfusion-induced kidney damage due to its antioxidant, anti-inflammatory and antiapoptotic properties.
    Keywords:  apoptosis; cytokines; evodiamine; kidney ischemia reperfusion; nucler factor-κB; oxidative stress
    DOI:  https://doi.org/10.1002/jcb.28976
  20. Biochem Biophys Res Commun. 2019 May 09. pii: S0006-291X(19)30855-1. [Epub ahead of print]
    Ting-Ting L, Yuan G, Jun L, Dan X, Hong-Bo T.
      Recent studies have implied that activation of AMP-dependent protein kinase (AMPK) could protect myocardial cells from oxygen glucose deprivation-re-oxygenation (OGD/R). The aim of the present study is to test whether GSK621, a novel and direct AMPK activator, could exert myocardial cell protection against OGD/R. We show that in AC16 human myocardial cells and primary murine myocardiocytes GSK621 dose-dependently activated AMPK signaling. GSK621 pretreatment potently inhibited OGD/R-induced viability reduction, cell death and apoptosis in AC16 cells and primary myocardiocytes. Furthermore, GSK621 attenuated OGD/R-induced reactive oxygen species production and oxidative injury in the myocardial cells. AMPKα1 knockdown (via targeted shRNA), knockout (via a CRISPR/Cas9 construct) or dominant negative mutation (T172A) not only blocked GSK621-induced AMPK activation, but also nullified GSK621-mediated myocardial cell protection against OGD/R. Further studies demonstrated that GSK621 activated AMPK downstream Nrf2 signaling. Contrarily, Nrf2 silencing by targeted shRNAs almost abolished GSK621-induced anti-OGD/R myocardial cell protection. We conclude that GSK621 protects myocardial cells from OGD/R through activation of AMPK-dependent signaling.
    DOI:  https://doi.org/10.1016/j.bbrc.2019.04.196
  21. Cell Prolif. 2019 May 16. e12627
    Xing JJ, Hou JG, Ma ZN, Wang Z, Ren S, Wang YP, Liu WC, Chen C, Li W.
      OBJECTIVES: Based on previous reports that ginsenosides have been shown to exert better preventive effects on cisplatin-induced kidney injury, the present work aims to evaluate the protective effects of ginsenoside Rb3 (G-Rb3) on cisplatin-induced renal damage and underlying mechanisms in vivo and in vitro.MATERIALS AND METHODS: The protective effect of G-Rb3 on cisplatin-induced acute renal failure in ICR mouse model and HEK293 cell model was investigated, and the underlying possible mechanisms were also explored. For animal experiment, renal function, kidney histology, inflammation, oxidative stress, relative protein molecules involved in apoptosis and autophagy signalling pathways were assessed. In addition, rapamycin (a specific inhibitor of mTOR), compound C (a specific inhibitor of AMPK) and acetylcysteine (NAC, a specific ROS scavenger) were employed to testify the effects of AMPK/mTOR signal pathway on the protective effects of G-Rb3 in HEK293 cells.
    RESULTS: Pre-treatment with G-Rb3 at doses of 10 and 20 mg/kg for ten days significantly reversed the increases in serum creatinine (CRE), blood urea nitrogen (BUN) and malondialdehyde (MDA), and decrease in glutathione (GSH) content and superoxide dismutase (SOD) activity. Histopathological examination further revealed that G-Rb3 inhibited cisplatin-induced nephrotoxicity. G-Rb3 diminished cisplatin-induced increase in protein expression levels of p62, Atg3, Atg5 and Atg7, and decrease in protein expression level of p-mTOR and the ratio of LC3-I/LC3-II, indicating that G-Rb3 suppressed cisplatin-induced activation of autophagy. Inhibition of autophagy induced inactivation of apoptosis, which suggested that autophagy played an adverse effect on cisplatin-evoked renal damage. Further, we found that G-Rb3 might potentially modulate the expressions of AMPK-related signal pathways.
    CONCLUSIONS: These findings clearly suggested that G-Rb3-mediated alleviation of cisplatin-induced nephrotoxicity was in part due to regulation of AMPK-/mTOR-mediated autophagy and inhibition of apoptosis in vitro and in vivo.
    Keywords:  AMPK/mTOR; Ginsenoside Rb3; HEK293 cells; autophagy; cisplatin; nephrotoxicity
    DOI:  https://doi.org/10.1111/cpr.12627
  22. Phytother Res. 2019 May 15.
    Wang Y, Yu C, Zhang H.
      Sinomenine (SIN) is an isoquinoline derived from Caulis Sinomenii that has been used to treat rheumatoid arthritis and osteoarthritis for several decades in China. This study aims to reveal the effects of SIN on mouse chondrogenic ATDC5 cells growth and inflammation. SIN was used to treat ATDC5 cells injured by lipopolysaccharides (LPS). The following parameters were determined for evaluating the treatment effects of SIN: cell viability, apoptosis, reactive oxygen species generation, and pro-inflammatory cytokines release. Besides, the expression of LPS-sensitive miRNA (miR-192) and the activation of NF-κB and MAPK signaling were studied to explain SIN's function. SIN with concentration of 30 μM significantly attenuated LPS-induced cell damage via increasing cell viability, inhibiting apoptosis and reactive oxygen species generation, and declining IL-6 and TNF-α release. miR-192 was downregulated by SIN treatment. Restoration of miR-192 expression by miRNA transfection could significantly impede SIN's protective action. Besides, the inhibitory effects of SIN on the activation of NF-κB and MAPK signaling were attenuated by miR-192 overexpression. Furthermore, GDF11 was found to be a target gene of miR-192. LPS-mediated injury to chondrogenic ATDC5 cells can be relieved by SIN via downregulating miR-192 and subsequently repressing the activation of NF-κB and MAPK signaling.
    Keywords:  Sinomenine; apoptosis; inflammation; lipopolysaccharides; miR-192; osteoarthritis
    DOI:  https://doi.org/10.1002/ptr.6372
  23. Am J Physiol Lung Cell Mol Physiol. 2019 May 15.
    Suryadevara V, Huang L, Kim SJ, Cheresh S, Shaaya M, Bandela M, Fu P, Feghali-Bostwick CA, Di Paolo G, Kamp DW, Natarajan V.
      Idiopathic pulmonary fibrosis (IPF) is a pernicious lung disease characterized by alveolar epithelial apoptosis, dysregulated repair of epithelial injury, scar formation and respiratory failure. In this study, we have identified phospholipase D (PLD) generated phosphatidic acid (PA) signaling in the development of pulmonary fibrosis (PF). Of the PLD isoenzymes, the protein expression of PLD2, but not PLD1, was up-regulated in lung tissues from IPF patients and bleomycin challenged mice. Both PLD1 (Pld1-/-) and PLD2 (Pld2-/-) deficient mice were protected against bleomycin induced lung inflammation and fibrosis, thereby establishing the role of PLD in fibrogenesis. The role of PLD1 and PLD2 in bleomycin-induced lung epithelial injury was investigated by infecting bronchial airway epithelial cells (Beas2B) with catalytically inactive mutants of PLD (hPLD1-K898R or mPld2-K758R), or down-regulation of expression of PLD1 or PLD2 with siRNA. Bleomycin stimulated mitochondrial (mt) superoxide production, mtDNA damage, and apoptosis in Beas2B cells, which was attenuated by the catalytically inactive mutants of PLD or PLD2 siRNA. These results show a role for PLD1 and PLD2 in bleomycin-induced generation of mt reactive oxygen species generation, mt DNA damage, and apoptosis of lung epithelial cells PF in mice. Thus, PLD may be a novel therapeutic target in ameliorating experimental PF in mice.
    Keywords:  Epithelial cell apoptosis; Mitochondrial DNA damage; Mitochondrial ROS; Phospholipase D2; Pulmonary Fibrosis
    DOI:  https://doi.org/10.1152/ajplung.00320.2018
  24. Iran J Pharm Res. 2019 ;18(1): 263-274
    Zangeneh F, Vazirizadeh A, Mirshamsi MR, Fakhri A, Faizi M, Pourahmad J.
      Despite recent improvements in treatment, ovarian cancer is still the leading cause of death from gynaecological malignancies. Today, marine mollusks are considered as natural source of new biologically and pharmacologically active compounds by scientists and the pharmaceutical industries. The aim of this study is to investigate the selective apoptotic effects of Turbo coronatus crude extract fractions on human epithelial ovarian cancer (EOC) cells and mitochondria. The cells and mitochondria were isolated from cancerous and non-cancerous ovarian tissues and exposed to IC50 concentration of F1 fraction for evaluation of mitochondrial and cellular parameters. Our results showed that F1 fraction of T. coronatus crude extract significantly induced toxic effects only in the cancerous ovarian mitochondria, including increased reactive oxygen species (ROS) formation, mitochondrial membrane depolarization, mitochondrial swelling, and cytochrome c release.Flow-cytometry analysis demonstrated that F1 fraction of T. coronatus progressively induced apoptosis and necrosis only on EOC but not non-cancerous cells. We eventuallyconcluded that F1 fraction of T. coronatus crude extract selectively induces apoptosis in EOC through a ROS- mediated pathway.
    Keywords:  Apoptosis; Epithelial ovarian cancer; F1 fraction; Gel filtration chromatography; Turbo coronatus
  25. Neurochem Res. 2019 May 17.
    Liu B, Liu W, Liu P, Liu X, Song X, Hayashi T, Onodera S, Ikejima T.
      Excessive physical exercise (overtraining; OT) increases oxidative stress and induces damage in multiple organs including the brain, especially the hippocampus that plays an important role in learning and memory. Silibinin, a natural flavonoid derived from milk thistle of Silybum marianum, has been reported to exert neuroprotective effect. In this study, rats were subjected to overtraining exercise, and the protective effects of silibinin were investigated in these models. Morris water maze and novel object recognition tests showed that silibinin significantly attenuated memory defects in overtrained rats. At the same time, the results of Nissl, TUNEL and SA-β-gal staining showed that silibinin reversed neuronal loss caused by apoptosis, and delayed cell senescence of the hippocampus in the overtrained rats, respectively. In addition, silibinin decreased malondialdehyde (MDA) levels which is associated with reactive oxygen species (ROS) generation. Silibinin prevented impairment of learning and memory caused by excessive physical exercise in rats, accompanied by reduced apoptosis and senescence in hippocampus cells.
    Keywords:  Apoptosis; Hippocampus; Overtraining; Senescence; Silibinin
    DOI:  https://doi.org/10.1007/s11064-019-02816-2
  26. Ann Otol Rhinol Laryngol. 2019 Jun;128(6_suppl): 117S-124S
    Park C, Lim H, Moon SK, Park R.
      OBJECTIVES: Auditory neuropathy due to toxicity mechanism of pyridoxine has not yet been fully documented. Therefore, the present study explored a direct mechanism underlying the effects of pyridoxine on auditory neuropathy in organ of Corti (OC) explants ex vivo and cochlear neuroblast cell line, VOT-33 in vitro.METHODS: Primary OC explants containing spiral ganglion neurons and cultured VOT-33 cells were treated with pyridoxine.
    RESULTS: In nerve fiber of primary OC explants, pyridoxine decreased staining for NF200, a neuro-cytoskeletal protein. We also found that pyridoxine-induced VOT-33 apoptosis, as indicated by accumulation of the sub-G0/G1 fraction, caspase-3 activation, and PARP cleavage. In addition, pyridoxine induced reactive oxygen species (ROS) generation and alteration of mitochondrial membrane potential transition (MPT), including Bcl-2 family protein expression and consequently Ca2+ accumulation and changes of endoplasmic reticulum (ER) stress-related protein expression such as phospho-PERK, caspase-12, Grp78, and CHOP.
    CONCLUSION: Pyridoxine preferentially induced severe cell death on nerve fiber in primary OC explants and markedly increased apoptotic cell death via mitochondria-mediated ER stress in VOT-33 cells.
    Keywords:  ER stress; VOT-33; auditory neuropathy; mitochondrial dysfunction; pyridoxine
    DOI:  https://doi.org/10.1177/0003489419836116
  27. Ecotoxicol Environ Saf. 2019 May 10. pii: S0147-6513(19)30548-2. [Epub ahead of print]180 168-178
    Jia Z, Wang H, Feng Z, Zhang S, Wang L, Zhang J, Liu Q, Zhao X, Feng D, Feng X.
      Fluorene-9-bisphenol (BHPF), a substitute for bisphenol A, is a chemical component of plastics for industrial production. There is evidence that BHPF exerts an antioestrogenic effect on mice, induces endometrial atrophy and leads to adverse pregnancy outcomes. However, the effects of BHPF on oocyte maturation and ovary development as well as its possible mechanisms remain unclear. The objective of this study was to investigate the toxicity and mechanism of BHPF exposure in mouse oocytes in vitro and in vivo. Our results showed that BHPF could inhibit the maturation of oocytes in vitro by reducing the protein level of p-MAPK and destroying the meiotic spindle. We found that in vitro, BHPF-treated oocytes showed increased ROS levels, DNA damage, mitochondrial dysfunction, and expression of apoptosis- and autophagy-related genes, such as Bax, cleaved-caspase 3, LC 3 and Atg 12. In addition, in vivo experiments showed that BHPF exposure could induce the expression of oxidative stress genes (Cat, Gpx 3 and Sod 2) and apoptosis genes (Bax, Bcl-2 and Cleaved-caspase 3) and increase the number of atresia follicles in the ovaries. Our data showed that BHPF exposure affected the first polar body extrusion of oocytes, increased oxidative stress, destroyed spindle assembly, caused DNA damage, altered mitochondrial membrane potentials, induced apoptosis and autophagy, and affected ovarian development.
    Keywords:  BHPF; Endocrine-disrupting chemicals; Oocyte maturation; Ovary damage; Oxidative stress
    DOI:  https://doi.org/10.1016/j.ecoenv.2019.05.019
  28. Artif Cells Nanomed Biotechnol. 2019 Dec;47(1): 1917-1923
    Li L, Lin G, Gu H, Yu L, Ni C.
      OBJECTIVE: To investigate the effect of Sulfiredoxin-1 (Srxn1) on astrocyte injury induced by hydrogen peroxide (H2O2).METHODS: Observing the changes of H2O2 on contents of lactate dehydrogenase (LDH), malondialdehyde (MDA), superoxide dismutase (SOD) and apoptosis after transfected Srxn1 siRNA into astrocytes. The protein expression of Notch 1, NICD and Hes1, the content of LDH and MDA, the activity of SOD and apoptosis rate of astrocytes after inhibiting or activation of Notch signalling pathway were detected by Western blot, ELISA and flow cytometry, respectively.
    RESULTS: Knockdown of Srxn1 could promote the secretion of LDH and MDA, decrease the activity of SOD and aggravate apoptosis of astrocytes induced by H2O2. The results of Western blot, ELISA assay and flow cytometry indicated that activation of the Notch signalling pathway attenuated the effect of Srxn1 on H2O2-induced oxidative damage and apoptosis of astrocytes.
    CONCLUSION: Srxn1 may protect astrocytes from oxidative stress injury induced by H2O2 by activation of Notch signalling pathway.
    Keywords:  Notch signalling pathway; Sulfiredoxin-1; apoptosis; astrocytes
    DOI:  https://doi.org/10.1080/21691401.2019.1614016
  29. Nano Lett. 2019 May 17.
    Liang S, Deng X, Chang Y, Sun C, Shao S, Xie Z, Xiao X, Ma P, Zhang H, Cheng Z, Lin J.
      As a noninvasive treatment modality, ultrasound (US)-triggered sonodynamic therapy (SDT) shows broad and promising applications to overcome the drawbacks of traditional photodynamic therapy (PDT) in combating cancer. However, the SDT efficacy is still not satisfactory without oxygen (O2) assistance. In addition, there is also much space to explore the SDT-based synergistic therapeutic modalities. Herein, a novel Pt-CuS Janus composed of hollow semiconductor CuS and noble metallic Pt was rationally designed and successfully synthesized. The hollow CuS shows a large inner cavity for loading sonosensitizer molecules (tetra-(4-aminophenyl) porphyrin, TAPP) to implement SDT. Moreover, the deposition of Pt not only enhances photothermal performance compared with those of CuS nanoparticles (NPs) due to the effect of the local electric field enhancement but also possesses nanozyme activity for catalyzing decomposition of endogenous overexpressed hydrogen peroxide (H2O2) to produce O2 that can overcome tumor hypoxia and augment the SDT-induced highly toxic reactive oxygen species (ROS) production for efficient cancer cell apoptosis. Importantly, the generated heat of Pt-CuS by 808 nm laser irradiation can accelerate the catalytic activity of Pt and elevate the O2 level that further facilitates SDT efficacy. Interestingly, the thermally sensitive copolymer coated around the Janus can act as a smart switch to regulate the catalytic ability of Pt and control TAPP release that has a significant effect on modulating the therapeutic effect. The synergistic catalysis-enhanced SDT efficiency and highly photothermal effect almost realized complete tumor resection without obvious reoccurrence and simultaneously displayed a highly therapeutic biosafety. Furthermore, the high optical absorbance allows the as-synthesized Pt-CuS Janus for photoacoustic (PA) imaging and NIR thermal imaging. This work develops a versatile nanoplatform for a multifunctional theranostic strategy and broadens the biological applications by rationally designing their structure.
    Keywords:  Pt-CuS Janus; Sonodynamic therapy; nanozymes; photothermal therapy; synergistic effect
    DOI:  https://doi.org/10.1021/acs.nanolett.9b01595
  30. Artif Cells Nanomed Biotechnol. 2019 Dec;47(1): 1938-1946
    Ke Y, Al Aboody MS, Alturaiki W, Alsagaby SA, Alfaiz FA, Veeraraghavan VP, Mickymaray S.
      Bionanotechnology has pivotal role in the development of a novel therapy, applications of gold nanoparticles (AuNPs) in the treatment of cancer. In this study, we found that therapeutics, pharmaceutics and diagnostic effectiveness of photosynthesized Catharanthus roseus (CR) AuNPs induces mitochondrial-mediated apoptotic signalling pathways via reactive oxygen species (ROS) induced cytotoxicity in cervical cancer cell line (HeLa) by in vitro model. The present examinations were for the most part centred around the gold chloride and photosynthesis AuNPs from the fluid leaf concentrate of CR and their harmful impacts on HeLa cell lines. The synthesized AuNPs were characterized using numerous biophysical analyses such as UV-vis, DLS, EDX, HR-TEM, SAED, FTIR and AFM. The synthesized AuNPs in the particle size range of 25-35 nm was confirmed by HR-TEM. The element gold and the crystalline nature of AuNPs were finalized using EDX, respectively. Anticancer potential of CR-AuNPs was studied using HeLa cells and the cytotoxic mechanism has been evaluated using MTT, mitochondrial-mediated apoptotic pathway through AO/EtBr staining assay, pro-apoptotic (Bax), anti-apoptotic (Bcl-2 and Bid) protein expression western blotting analysis and caspases activity using ELISA analysis. In in vitro study, the IC50 of HeLa cells was found to be 5 µg/ml confirmed using MTT assay. The present data revealed that drug delivery vehicles developed on CR-AuNPs nanocomplexes might include extensive purpose in human cancer diagnosis and treatment.
    Keywords:  ; Gold nanoparticles; apoptosis; cervical cancer; nanotechnology
    DOI:  https://doi.org/10.1080/21691401.2019.1614017
  31. Skin Pharmacol Physiol. 2019 May 10. 1-9
    Kahremany S, Babaev I, Gvirtz R, Ogen-Stern N, Azoulay-Ginsburg S, Senderowitz H, Cohen G, Gruzman A.
      BACKGROUND/AIMS: The Nrf2 signaling pathway plays a pivotal role in neutralizing excess reactive oxygen species formation and therefore enhancing the endogenous cellular protection mechanism. Thus, activating this pathway may provide therapeutic options against oxidative stress-related disorders. We have recently applied a computer-aided drug design approach to the design and synthesis of novel Nrf2 enhancers. The current study was aimed at investigating the potential beneficial impact of (E)-5-oxo-1-(4-((2,4,6-trihydroxybenzylidene)amino)phenyl)pyrrolidine-3-carboxylic acid (SK-119) in skin oxidative damage models.METHODS: SK-119, tested initially in PC-12 cells, attenuated oxidative stress-induced cytotoxicity concomitantly with Nrf2 activation. The potential impact of this compound was evaluated in skin-based disease models both in vitro (HaCaT cells) and ex vivo (human skin organ culture).
    RESULTS: The data clearly showed the marked anti-inflammatory and photoprotection properties of the compound; SK-119-treated cells or tissues displayed a reduction in cytokine secretion induced by lipopolysaccharides (LPS) in a manner comparable with dexamethasone. In addition, topical application of SK-119 was able to block UVB-induced oxidative stress and attenuated caspase-mediated apoptosis, DNA adduct formation, and the concomitant cellular damage.
    CONCLUSION: These results indicate that SK-119 is an Nrf2 activator that can be used as a prototype molecule for the development of novel treatments of dermatological disorders related to oxidative stress.
    Keywords:  LPS-induced damage; Nrf2 activator; SK-119; UVB
    DOI:  https://doi.org/10.1159/000499432
  32. Oxid Med Cell Longev. 2019 ;2019 4353791
    Yang XX, Liang L, Liu X, Li FJ, Dong JC, Mu JK, Zeng LX, Bi Q, Gu W, Yu J.
      Targeting mitochondria as a hepatic-protective strategy has gained attention, because of their important roles in energy production, adjustment of apoptosis, and generation of reactive oxygen species. To promote the discovery of natural mitochondria-targeted hepatic-protectants, we established a hepatocellular mitochondria-based capturing method by coupling affinity ultrafiltration with liquid chromatography/mass spectrometry (LC/MS), which is suitable for identifying mitochondrial ligands from medicinal herbs (MHs). After evaluating the feasibility of the method, it was applied for capturing mitochondria-targeting constituents from Peucedani Radix extract. A total of 10 active compounds were identified by LC/MS, all of which were newly identified mitochondrial ligands. The mitochondria-remedying activity of 4 of the 10 hits was confirmed by pharmacological tests in vitro. Additionally, the hepatic-protective abilities of 4 hits were verified in both carbon tetrachloride-damaged liver L02 cells and mice. These results indicated that the method could be used for identifying hepatic mitochondria-targeting constituents in MHs, which might be beneficial for hepatic-protective development.
    DOI:  https://doi.org/10.1155/2019/4353791
  33. Cell Death Dis. 2019 May 15. 10(6): 376
    Slobodnyuk K, Radic N, Ivanova S, Llado A, Trempolec N, Zorzano A, Nebreda AR.
      Apoptosis and senescence are two mutually exclusive cell fate programs that can be activated by stress. The factors that instruct cells to enter into senescence or apoptosis are not fully understood, but both programs can be regulated by the stress kinase p38α. Using an inducible system that specifically activates this pathway, we show that sustained p38α activation suffices to trigger massive autophagosome formation and to enhance the basal autophagic flux. This requires the concurrent effect of increased mitochondrial reactive oxygen species production and the phosphorylation of the ULK1 kinase on Ser-555 by p38α. Moreover, we demonstrate that macroautophagy induction by p38α signaling determines that cancer cells preferentially enter senescence instead of undergoing apoptosis. In agreement with these results, we present evidence that the induction of autophagy by p38α protects cancer cells from chemotherapy-induced apoptosis by promoting senescence. Our results identify a new mechanism of p38α-regulated basal autophagy that controls the fate of cancer cells in response to stress.
    DOI:  https://doi.org/10.1038/s41419-019-1607-0
  34. Hum Exp Toxicol. 2019 May 12. 960327119846941
    Moon SH, Lee CM, Nam MJ.
      Cadmium (Cd) is a heavy metal widely used in industry, and the skin is an important target of this metal. Taxifolin (Tax), a natural source of bioflavonoids found in various conifers, exerts multiple biologic effects on skin cells. However, the mechanisms by which Tax protects keratinocytes against Cd are currently unclear. We investigated the cytoprotective effects of Tax against Cd-induced apoptosis in the human HaCaT keratinocyte. The water-soluble tetrazolium salt (WST-1) assay and Annexin V/propidium iodide double-staining assay results showed that Cd-induced cell death was lower in cells treated with Tax (0-100 μM) than in cells treated with Cd alone. Additionally, a reduction of Cd-induced DNA fragmentation by Tax was shown by terminal deoxynucleotidyl transferase (TdT)-mediated dUTP nick-end labeling assay. The levels of reactive oxygen species were also lower in Cd/Tax-treated cells than in Cd-treated cells. We employed a two-dimensional electrophoresis-based proteomic analysis to identify treatment-related alterations in protein expression. Tax downregulated cathepsin B and D and upregulated hsp27, cyclophilin A, and peroxiredowin-1. Western blotting confirmed the downregulation of cathepsin B and D and the upregulation of hsp27. The cytoprotective effects of Tax against Cd-induced apoptosis were also characterized by the changes in the activity of caspase 3, -7, poly ADP-ribose polymerase, the cellular proliferation-related ERK1/2, and AKT. Furthermore, the levels of cell cycle-related proteins, such as SP1 and p21, decreased, whereas p53 level increased. We concluded that Tax reduced Cd cytotoxicity and Cd-induced apoptosis by inhibiting the apoptotic pathway.
    Keywords:  HaCaT; Taxifolin; cadmium; toxicity
    DOI:  https://doi.org/10.1177/0960327119846941
  35. Cell Cycle. 2019 May 16.
    Bao Z, Dai X, Wang P, Tao Y, Chai D.
      An accumulating body of evidence has shown that capsaicin induces apoptosis in various tumor cells as a mechanism of its anti-tumor activity. However, the effects of capsaicin on osteosarcoma have not been studied extensively. In the current study, we explore the molecular mechanism of capsaicin-mediated tumor suppressive function in osteosarcoma. We found that capsaicin induced apoptosis and the activation of transient receptor potential receptor vanilloid 1 (TRPV1) in a dose- and time-dependent manner in human osteosarcoma MG63 cells in vitro. Blocking TRPV1 using capsazepine attenuated the capsaicin-induced cytotoxicity, mitochondrial dysfunction, overproduction of reactive oxygen species (ROS) and decrease in superoxide dismutase (SOD) activity. In addition, the results demonstrated that capsaicin induced the activation of adenosine 5'-monophosphate-activated protein kinase (AMPK), p53 and C-jun N-terminal kinase (JNK). In addition, Compound C (antagonist of AMPK) attenuated the activation of p53, which appeared to be TRPV1 independent. Taken together, the present study suggests that capsaicin effectively causes cell death in human osteosarcoma MG63 cells via the activation of TRPV1-dependent (mitochondrial dysfunction, and overproduction of ROS and JNK) and TRPV1-independent (AMPK-p53) pathways. Thus, capsaicin may be a potential anti-osteosarcoma agent.
    Keywords:  Apoptosis; Capsaicin; Mitochondria; Osteosarcoma; TRPV1
    DOI:  https://doi.org/10.1080/15384101.2019.1618119
  36. Cancers (Basel). 2019 May 11. pii: E656. [Epub ahead of print]11(5):
    Eberle J.
      Melanoma of the skin has become a prime example for demonstrating the success of targeted cancer therapy. Nevertheless, high mortality has remained, mainly related to tumor heterogeneity and inducible therapy resistance. But the development of new therapeutic strategies and combinations has raised hope of finally defeating this deadly disease. TNF-related apoptosis-inducing ligand (TRAIL) represents a promising antitumor strategy. The principal sensitivity of melanoma cells for TRAIL was demonstrated in previous studies; however, inducible resistance appeared as a major problem. To address this issue, combination strategies were tested, and survival pathway inhibitors were shown to sensitize melanoma cells for TRAIL-induced apoptosis. Finally, cell cycle inhibition was identified as a common principle of TRAIL sensitization in melanoma cells. Mitochondrial apoptosis pathways, pro- and antiapoptotic Bcl-2 proteins as well as the rheostat consisted of Smac (Second mitochondria-derived activator of caspase) and XIAP (X-linked inhibitor of apoptosis protein) appeared to be of particular importance. Furthermore, the role of reactive oxygen species (ROS) was recognized in this setting. Inducible TRAIL resistance in melanoma can be explained by (i) high levels of antiapoptotic Bcl-2 proteins, (ii) high levels of XIAP, and (iii) suppressed Bax activity. These hurdles have to be overcome to enable the use of TRAIL in melanoma therapy. Several strategies appear as particularly promising, including new TRAIL receptor agonists, Smac and BH3 mimetics, as well as selective kinase inhibitors.
    Keywords:  Bax; Bcl-2 proteins; Smac; TRAIL; XIAP; kinases; melanoma
    DOI:  https://doi.org/10.3390/cancers11050656
  37. Photodiagnosis Photodyn Ther. 2019 May 09. pii: S1572-1000(18)30367-3. [Epub ahead of print]
    Postigo A, Schiavi PC, Funes M, Sortino M.
      Porophyllum obscurum (Spreng) DC (Asteraceae) hexanic extract (PoHex) from aerial parts has demonstrated antifungal activity under UVA irradiation against Candida spp. isolates from patients with oropharyngeal candidiasis and four thiophenes were isolated as responsible of the activity. In the present work, we studied the photomechanisms whereby PoHex and their thiophenes produce photoinactivation of C. albicans. Reactive Oxygen Species generation by PoHex and thiophenes was evaluated: the production of superoxide anion, employing the NBT reduction assay; hydrogen peroxide, through the formation of a red quinoneimine; and singlet oxygen by using the 1,3-DPBF bleaching method. The action of ROS in fungal cells was investigated by evaluating binding of photosensitizer, leakage, apoptosis and stress sensibility that were performed by following M27-A3 guidelines, in parallel under "light" and "darkness" conditions. Results showed that the photosensitive antifungal activity of PoHex required oxygen and both type I (production of superoxide anion and hydrogen peroxide) and type II (production of singlet oxygen) reactions were involved. In addition, we found that ROS generated by PoHex did not cause release of cytoplasmic components due to membrane damage nor apoptosis of C. albicans. Treatment with PoHex and UVA increased cells sensitivities to osmotic stressors; did not reduce resistance to additional oxidative stress and possibly affected the structure of the cell wall. In addition, 2,2':5'2″terthiophene, the most active PS present in PoHex and the only one that generate single oxygen, at Minimal Fungicide Concentration, did not cause leakage nor apoptosis and did not increase sensitivities to osmotic and oxidative stressors. Results demonstrated that Photodynamic Inactivation employing PoHex under UVA does represent an alternative for topical antifungal therapy for oropharyngeal candidiasis.
    Keywords:  Candida albicans; Porophyllum obscurum; photodynamic therapy; reactive oxygen species
    DOI:  https://doi.org/10.1016/j.pdpdt.2019.05.003
  38. Oxid Med Cell Longev. 2019 ;2019 1729013
    Huang S, Zheng B, Jin X, Yu Q, Zhang X, Sun X, Chen Y, Ren X, Wismeijer D, Ma J, Zhang C, Wu G, Pan Y.
      Pathological stimuli, such as bacterial activity, dental bleaching, and nonpolymerized resin monomers, can cause death of dental pulp cells (DPCs) through oxidative stress- (OS-) induced mitochondrial dysfunction. However, the crucial molecular mechanisms that mediate such a phenomenon remain largely unknown. OS is characterized by the overproduction of reactive oxygen species (ROS), e.g., H2O2, O2 -, and ·OH. Mitochondria are a major source of ROS and the principal attack target of ROS. Cyclophilin D (CypD), as the only crucial protein for mitochondrial permeability transition pore (mPTP) induction, facilitates the opening of mPTP and causes mitochondrial dysfunction, leading to cell death. In the present study, we hypothesized that CypD-mediated mitochondrial molecular pathways were closely involved in the process of OS-induced death of human DPCs (HDPCs). We tested the phenotypic and molecular changes of HDPCs in a well-established OS model-H2O2 treatment. We showed that H2O2 dramatically reduced the viability and increased the death of HDPCs in a time- and dose-dependent manner by performing MTT, flow cytometry, and TUNEL assays and quantifying the expression changes of Bax and Bcl-2 proteins. H2O2 also induced mitochondrial dysfunction, as reflected by the increased mitochondrial ROS, reduced ATP production, and activation of mPTP (decreased mitochondrial membrane potential and enhanced intracellular Ca2+ level). An antioxidant (N-acetyl-L-cysteine) effectively preserved mitochondrial function and significantly attenuated H2O2-induced cytotoxicity and death. Moreover, H2O2 treatment markedly upregulated the CypD protein level in HDPCs. Notably, genetic or pharmacological blockade of CypD significantly attenuated H2O2-induced mitochondrial dysfunction and cell death. These findings provided novel insights into the role of a CypD-dependent mitochondrial pathway in the H2O2-induced death in HDPCs, indicating that CypD may be a potential therapeutic target to prevent OS-mediated injury in dental pulp.
    DOI:  https://doi.org/10.1155/2019/1729013
  39. Acta Biomater. 2019 May 09. pii: S1742-7061(19)30323-X. [Epub ahead of print]
    Sun B, Chen Y, Yu H, Wang C, Zhang X, Zhao H, Chen Q, He Z, Luo C, Sun J.
      The combination of chemotherapy with photodynamic therapy (PDT) holds promising applications in cancer therapy. However, co-encapsulation of chemotherapeutic agents and photosensitizers (PS) into the conventional nanocarriers suffers from inefficient co-loading and aggregation-caused quenching (ACQ) effect of PS trapped in dense carrier materials. Herein, we report a light-activatable photodynamic PEG-coated prodrug nanoplatform for core-shell synergistic chemo-photodynamic therapy. A novel photodynamic polymer is rationally designed and synthesized by conjugating pyropheophorbide a (PPa) to polyethylene glycol 2000 (PEG2k). PPa is used as the hydrophobic and photodynamic moiety of the amphipathic PPa-PEG2k polymer. Then, a core-shell nanoassembly is prepared, with an inner core of a reactive oxygen species (ROS)-responsive oleate prodrug of paclitaxel (PTX) and an outer layer of PPa-PEG2k. PPa-PEG2k serves for both PEGylation and PDT. Instead of being trapped in the inner core, PPa in the outer PPa-PEG2k layer significantly alleviates the ACQ effect. Under laser irradiation, ROS generated by PPa-PEG2k not only is used for PDT but also synergistically promotes PTX release in combination with the endogenous ROS overproduced in tumor cells. The photodynamic PEG-coated nanoassemblies demonstrated synergistic antitumor activity in vivo. Such a unique nanoplatform, with an inner chemotherapeutic core and an outer photodynamic PEG shell, provides a new strategy for synergistic chemo-photodynamic therapy. STATEMENT OF SIGNIFICATION: The combination of chemotherapy with photodynamic therapy (PDT) holds promising prospects in cancer therapy. However, it remains a tremendous challenge to effectively co-deliver chemotherapeutic drugs and photosensitizers into tumors. Herein, we construct a photodynamic PEGylation-coated prodrug-nanoplatform for high-efficiency synergistic cancer therapy, which is composed of a light-activatable PPa-PEG2k shell and a ROS-responsive paclitaxel (PTX) prodrug core. The PPa-PEG2k-generated ROS not only was used for synergistic PTX release but also synergistically facilitated tumor cell apoptosis in combination with PTX-initiated chemo-cytotoxicity. The light-activatable nanoassemblies exhibited multiple drug delivery advantages including high co-loading efficiency, self-enhanced PTX release, extended circulation time, favorable biodistribution, and potent synergistic anticancer activity. Our findings provide a new strategy for the rational design of advanced nano-DDS for high-efficiency combinational chemo-photodynamic therapy.
    Keywords:  Chemo-photodynamic therapy; Core-shell nanoassembly; Light-activatable; Photodynamic PEGylation; Synergistic
    DOI:  https://doi.org/10.1016/j.actbio.2019.05.008
  40. Toxicol In Vitro. 2019 May 08. pii: S0887-2333(18)30690-8. [Epub ahead of print]
    Ji S, Sun R, Xu K, Man Z, Ji J, Pu Y, Yin L, Zhang J, Pu Y.
      Prodigiosin contains a tripyrrole skeleton and shows impressive anticancer potential in multiple cell lines. Numerous studies have been conducted on prodigiosin-induced apoptosis and the related mechanisms. However, few reports have considered the effects of prodigiosin on autophagy and the relationship between apoptosis and autophagy. Here, we examined whether prodigiosin affected apoptosis and autophagy through the extracellular signal-regulated (ERK) signaling pathway in K562 cells, employing cell proliferation, flow cytometry, caspase activity, and western blot analyses. Inhibition of the ERK signaling pathway with PD184352 was conducted to verify the role of this pathway on prodigiosin-mediated processes. Our findings revealed that prodigiosin inhibited the proliferation of K562 cells, increased reactive oxygen species (ROS), induced apoptosis and inhibited autophagy in K562 cells. Additionally, the ROS scavenger, N-Acetyl-L-cysteine (NAC), partially prevented prodigiosin-induced apoptosis but did not reduce prodigiosin-inhibited autophagy in K562 cells. Furthermore, prodigiosin treatment in K562 cells reduced the phosphorylation of c-Jun N-terminal kinases (JNKs) and P38, and activated ERK signaling pathway. When ERK1/2 phosphorylation was blocked by PD184352, prodigiosin-induced apoptosis and the inhibition of autophagy decreased significantly. Taken together, these results demonstrated that the ERK signaling pathway was involved in prodigiosin-induced apoptosis and prodigiosin-inhibited autophagy.
    Keywords:  Apoptosis; Autophagy; ERK signaling pathway; K562 cells; Prodigiosin
    DOI:  https://doi.org/10.1016/j.tiv.2019.05.003
  41. Cell Mol Biol (Noisy-le-grand). 2019 Apr 30. 65(4): 101-106
    Wang M, Xia L, Fu G.
      Endothelial progenitor cells (EPCs) improve neovascularization and endothelium regeneration. Transplantation with EPCs is a therapeutic strategy for the treatment of ischemic diseases. However, the transplanted EPCs are susceptible to adverse environments such as hypoxia, inflammation and oxidative stress. Oxidative stress-induced apoptosis of transplanted EPCs greatly reduces their therapeutic efficacy. Lipopolysaccharide (LPS) is a highly immunogenic antigen. Recent findings suggest that low dose of LPS pretreatment has protective effect against apoptosis. In this study, the role of LPS in apoptosis of EPCs was investigated. Pretreatment with 1µg/ml LPS prevented oxidative stress-induced EPCs apoptosis and ROS generation, which effects were abolished by TAK-242, a specific TLR4 antagonist. Further investigation of the mechanisms demonstrated that the activation was mediated by TLR4, and that PI3K/Akt/ NF-κB p65 signaling pathway may play a critical role in the process.
    Keywords:  Endothelial progenitor cells; Lipopolysaccharide; Oxidative stress; Toll-like receptors 4.
  42. J Cell Biochem. 2019 May 17.
    Fan Y, Zhang Y, Liu Y, Xu W, Yang Y, Hao Y, Tao L.
      Cervical cancer is the fourth most lethal human malignancy and the leading cause of death among females around the world. Many antitumor agents have microbial origins. 5'-epi-SPA-6952A is a new 24-membered macrolide isolated from the cultured broth of Streptomyces diastatochromogenes. Therefore, we studied the activity and molecular mechanism of 5'-epi-SPA-6952A in human cervical carcinoma HeLa cell. The results showed that 5'-epi-SPA-6952A significantly inhibited cell proliferation and migration. In addition, 5'-epi-SPA-6952A obviously increased the production of intracellular reactive oxygen species and DNA damage in HeLa cells. Moreover, nuclear shrinkage of cells, decrease in mitochondrial membrane potential, and upregulation of Bax/Bcl-2 ratio resulted in the release of cytochrome c, and activation of caspase-9/3 was observed in HeLa cells treated with 5'-epi-SPA-6952A, which means it enhanced the intrinsic mitochondrial apoptosis. Besides, DNA-damage associated proteins poly (ADP-ribose) polymerase (PARP) and p53 were also studied, and the expressions of cleaved-PARP and p53 were drastically increased in HeLa cells treated with 5'-epi-SPA-6952A. Furthermore, we confirmed that 5'-epi-SPA-6952A affected the survival of HeLa cells by blocking cell cycle progression in the G1 phase. Taken together, the results shows that 5'-epi-SPA-6952A significantly inhibited HeLa cells proliferation via intrinsic mitochondrial apoptosis, cell cycle arrest, and blocking cell migration.
    Keywords:  DNA damage; anticervical cancer activity; cell cycle arrest; cell migration; mitochondria/caspase-mediated apoptosis; natural product
    DOI:  https://doi.org/10.1002/jcb.28939
  43. Cell Biochem Biophys. 2019 May 14.
    Mast JM, Tse D, Shee K, Lakshmi Kuppusamy M, Kmiec MM, Kálai T, Kuppusamy P.
      Pancreatic cancer has a 5-year survival rate below 10% and the treatment options are limited. Signal transducer and activator of transcription (STAT3) is a constitutively expressed protein in human pancreatic cancers and is associated with their poor prognosis. Targeting of STAT3 signaling using novel therapeutic agents is a potential strategy for pancreatic cancer treatment. Diarylidenylpiperidone (DAP) compounds, such as H-4073 and HO-3867, have been shown to be STAT3 inhibitors in several human ovarian cancers. Particularly, HO-3867 is an N-hydroxypyrroline derivative of DAP that has targeted cytotoxicity toward cancer cells without affecting healthy cells. In the present study, we evaluated the anticancer efficacy of H-4073 and HO-3867 in a human pancreatic cell line (AsPC-1). We found that both the compounds exhibited potential cytotoxicity to AsPC-1 cells by inducing G2/M cell-cycle arrest, apoptosis, and cell death, by mitochondrial damage and inhibition of STAT3 phosphorylation. In summary, H-4073 and HO-3867 are cytotoxic to AsPC-1 cells and seem to act through similar mechanisms, including STAT3 inhibition, cell-cycle arrest, and apoptosis.
    Keywords:  Apoptosis; Diarylidenylpiperidone; Nitroxide; Pancreatic cancer; Reactive oxygen species; STAT3
    DOI:  https://doi.org/10.1007/s12013-019-00873-6
  44. Oxid Med Cell Longev. 2019 ;2019 3501059
    Jiao Y, Niu T, Liu H, Tay FR, Chen JH.
      Dental resin monomers such as 2-hydroxyethyl methacrylate (HEMA) disturb vital cell functions and induce mitochondrial intrinsic apoptosis via generation of oxidative stress. Nuclear factor erythroid 2-related factor 2 (Nrf2) regulates the gene expression of antioxidative enzymes and plays a crucial role in the maintenance of cellular redox equilibrium and mitochondrial homeostasis. The present study investigated the functional significance of Nrf2 in cellular response toward HEMA. It was found that HEMA stimulation promoted nuclear translocation of Nrf2 and increased Nrf2 and heme oxygenase-1 (HO-1) expression, which was further enhanced by Nrf2 activator tert-butylhydroquinone (tBHQ), but suppressed by Nrf2 inhibitor ML385. Pretreatment of primary human dental pulp cells (hDPCs) with tBHQ protected the cells from HEMA-induced oxidative injury (increased reactive oxygen species production and apoptosis) and mitochondrial impairment (morphological alterations, decreased ATP production, suppressed oxidative phosphorylation activity, depolarization of mitochondrial membrane potential, and disrupted electron transport chain). In contrast, pretreatment with ML385 increased cell sensitivity to these injurious processes. This protective effect on mitochondria could be related to peroxisome proliferator-activated receptor γ coactivator 1α (PGC1α)/nuclear respiratory factor 1 (NRF1) pathway. These results contribute to the understanding of the function of Nrf2 and the development of novel therapies to counteract the adverse effects of dental resin monomers.
    DOI:  https://doi.org/10.1155/2019/3501059
  45. Aging (Albany NY). 2019 May 14.
    Song H, Zuo Z, Yang Z, Gao C, Chen K, Fang J, Cui H, Ouyang P, Deng J, Geng Y, Guo H.
      This research was to investigate the difference of hepatic histopathology and apoptosis between the diet-induced obesity (DIO) and normal (lean) mice after Escherichia coli (E. coli) pneumonia. A total of 128 ICR mice were selected to be challenged intranasally with phosphate-buffered saline (PBS) or 4×109CFUs/mL of E. coli, and the liver histopathology and apoptosis were examined pre- and post-infection. Results showed that the liver index, levels of lipid droplets, cytokines, adipocytokines, oxidative stress, apoptotic percentage, and apoptotic related factors in the E. coli-infected mice were generally higher than those in the uninfected mice, whereas the hepatic glycogen and Bcl-2 were the opposite. Interestingly, after E. coli infection, the DIO-E. coli mice exhibited decreased liver index and apoptotic percentages, and reduced levels of TNF-α, IL-6, resistin, MDA, GSH, CAT, Caspase-3, Caspase-9, Bax as well as Bax/Bcl-2 ratio in comparison to the lean-E. coli mice. Our results indicated that E. coli-induced pneumonia caused hepatic histopathological damage, increased hepatic apoptosis, oxidative damages, and higher levels of cytokines and adipocytokines. However, such changes showed less severely in the DIO mice than in the lean mice following E. coli pneumonia.
    Keywords:  Escherichia coli-pneumonia; apoptosis; histopathology; liver; obesity
    DOI:  https://doi.org/10.18632/aging.101956
  46. Nutrients. 2019 May 15. pii: E1082. [Epub ahead of print]11(5):
    Ikram M, Saeed K, Khan A, Muhammad T, Khan MS, Jo MG, Rehman SU, Kim MO.
      The aim of the current study was to explore the underlying neuroprotective mechanisms of curcumin (50 mg/kg, for six weeks) against ethanol (5 mg/kg i.p., for six weeks) induced oxidative stress and inflammation-mediated cognitive dysfunction in mice. According to our findings, ethanol triggered reactive oxygen species (ROS), apoptosis, neuroinflammation, and memory impairment, which were significantly inhibited with the administration of curcumin, as assessed by ROS, lipid peroxidation (LPO), and Nrf2/HO-1 (nuclear factor erythroid 2-related factor 2/Heme-oxygenase-1) expression in the experimental mice brains. Moreover, curcumin regulated the expression of the glial cell markers in ethanol-treated mice brains, as analyzed by the relative expression TLR4 (Toll like Receptor 4), RAGE (Receptor for Advanced Glycations End products), GFAP (Glial fibrillary acidic protein), and Iba-1 (Ionized calcium binding adaptor molecule 1), through Western blot and confocal microscopic analysis. Moreover, our results showed that curcumin downregulated the expression of p-JNK (Phospo c-Jun N-Terminal Kinase), p-NF-kB (nuclear factor kappa-light-chain-enhancer of activated B cells), and its downstream targets, as assessed by Western blot and confocal microscopic analysis. Finally, the expression of synaptic proteins and the behavioral results also supported the hypothesis that curcumin may inhibit memory dysfunction and behavioral alterations associated with ethanol intoxication. Altogether, to the best of our knowledge, we believe that curcumin may serve as a potential, promising, and cheaply available neuroprotective compound against ethanol-associated neurodegenerative diseases.
    Keywords:  apoptosis; neurodegenerative diseases; neuroinflammation; oxidative stress; synaptic dysfunction
    DOI:  https://doi.org/10.3390/nu11051082
  47. Neurochem Int. 2019 May 08. pii: S0197-0186(19)30163-9. [Epub ahead of print]
    Lee SY, Hur SJ.
      The protective effect of two 3 kDa peptide fractions (AK3KF1 and AK3KF2), obtained from beef myofibrillar protein using an inexpensive enzyme (alkaline-AK) on human neuronal cells (SH-SY5Y) against H2O2-induced apoptosis was investigated. These peptides were isolated and further separated by fast protein liquid chromatography (FPLC), and their protective effect against H2O2-mediated cell death was measured by determining cell viability, nitric oxide (NO) production, mitochondrial membrane potential (MMP), apoptosis, morphological changes in cell nuclei, and in vitro antioxidant assays. The results indicated that treatment with peptide fractions increased cell viability and MMP, and decreased NO production, fragmentation of cell nuclei, and apoptosis in H2O2-treated SH-SY5Y cells. This is the first study to report neuroprotective effects of a peptide obtained from beef myofibrillar protein. The peptide sequence was identified as Thr-Gln-Lys-Lys-Val-Ile-Phe-Cys (TQKKVIFC). Thus, these findings suggest that TQKKVIFC can prevent neuronal cell death and could be useful in preventing neurodegenerative diseases.
    Keywords:  Beef; Cell death; Neuroprotective effect; Peptide; SH-SY5Y cells
    DOI:  https://doi.org/10.1016/j.neuint.2019.05.003
  48. J Cell Biochem. 2019 May 14.
    Ma C, Chen J, Li P.
      Rheumatoid arthritis (RA) is an autoimmune disease of the joints characterized by synovial hyperplasia and chronic inflammation. Fibroblasts-like synoviocytes (FLS), major cells in the synovium, together with infiltrated leukocytes, contribute greatly to RA progression. In our study, we hypothesized that geldanamycin (GA), a cancer drug might be able to inhibit RA FLS growth. To test the idea, RA FLS were isolated and cultured for cancer drug test. The results showed that GA can specifically inhibit the growth of RA FLS compared with normal FLS. Essentially, GA was found to promote reactive oxygen species production in RA FLS and induce programmed cell death. The annexinV/propidium iodide and terminal deoxynucleotidyl transferase dUTP nick-end labeling staining confirmed that GA can directly induce apoptosis and subsequently inhibit the growth of RA FLS, which was also confirmed by Western blot assay. In addition, our data demonstrated that inflammation was inhibited by suppressed nuclear factor κB signaling pathway. The therapeutic effect of GA was explored in collagen-induced arthritis mice. In short, GA was a promising drug for the treatment of RA by specifically inhibiting the proliferation and inflammation of RA FLS.
    Keywords:  apoptosis; fibroblast-like synoviocytes; geldanamycin; inflammation; rheumatoid arthritis
    DOI:  https://doi.org/10.1002/jcb.28906
  49. Toxicol Appl Pharmacol. 2019 May 11. pii: S0041-008X(19)30167-X. [Epub ahead of print]
    Yang W, Zhang G, Jiang F, Zeng Y, Zou P, An H, Chen Q, Ling X, Han F, Liu W, Yang H, Liu J, Cao J, Ao L.
      There is increasing evidence that indicates benzo[a]pyrene (B[a]P) and its active metabolite benzo[a]pyrene-7, 8-dihydrodiol-9, 10-epoxide (BPDE) are endocrine disruptors that can cause reproductive toxicity. Nevertheless, the underlying mechanisms are still obscure. The present study investigates the impacts of B[a]P and BPDE on mitochondria, a sensitive target affected by multiple chemicals, in spermatogenic cells. It showed that BPDE treatment induced mitochondrial dysfunction and the inhibition of mitochondrial biogenesis in mouse spermatocyte-derived cells (GC-2). These effects were efficiently mitigated by pretreatment with ZLN005, an activator of PGC-1α, in GC-2 cells. TERT knockdown and re-expression cell models were established to demonstrate that TERT regulated the BPDE-induced mitochondrial damage via PGC-1α signaling in GC-2 cells. Moreover, upregulating or knockdown SIRT1 expression attenuated or aggravated BPDE-induced mitochondrial compromise by activating or inhibiting, respectively, the TERT and PGC-1α molecules in GC-2 cells. Finally, we observed that BPDE markedly elevated oxidative stress in GC-2 cells. Resveratrol and N-acetylcysteine, as reactive oxygen species (ROS) scavengers, attenuated BPDE-mediated mitochondrial damage by increasing SIRT1 activity and expression in GC-2 cells. The in vitro results were corroborated by in vivo experiments in rats treated with B[a]P for 4 weeks. B[a]P administration caused mitochondrial damage and mitochondria-dependent apoptosis in spermatogenic cells, as well as the decreased expression of SIRT1, TERT, and PGC-1α. In summary, the results of the present study demonstrate that B[a]P and BPDE induce mitochondrial damage through ROS production that suppresses SIRT1/TERT/PGC-1a signaling and mediate B[a]P- and BPDE-mediated reproductive toxicity.
    Keywords:  Benzo[a]pyrene; Benzo[a]pyrene-7,8-dihydrodiol-9,10-epoxide; Male reproductive toxicity; Mitochondrial damage; Reactive oxygen species
    DOI:  https://doi.org/10.1016/j.taap.2019.05.004
  50. Chemosphere. 2019 May 03. pii: S0045-6535(19)30914-2. [Epub ahead of print]229 314-323
    Hamdi H, Abid-Essefi S, Eyer J.
      Epoxiconazole (EPX) is a very effective fungicide of the triazole family. Given its wide spectrum of use, the increased application of this pesticide may represent a serious risk on human health. Previous studies have found that EPX is cytotoxic to cells, although the exact mechanism remains elusive. In particular, the effect on the nervous system is poorly elucidated. Here we evaluated the implication of oxidative stress in the neurotoxicity and studied its apoptotic mechanism of action. We demonstrated that the treatment by EPX reduces the viability of cells in a dose dependent manner with an IC50 of 50 μM. It also provokes the reduction of cell proliferation. EPX could trigger arrest in G1/S phase of cell cycle with low doses, however with IC50, it induced an accumulation of F98 cells in G2/M phase. Moreover, EPX induced cytoskeleton disruption as evidenced by immunocytochemical analysis. It provoked also DNA fragmentation in a concentration dependent manner. The EPX induced apoptosis, which was observed by morphological changes and by positive Annexin V FITC-PI staining concurrent with a depolarization of mitochondria. Furthermore, the cell mortality provoked by EPX was significantly reduced by pretreatment with Z-VAD-FMK, a caspase inhibitor. Moreover, N-acetylcysteine (NAC) strongly restores cell viability that has been inhibited by EPX. The results of these findings highlight the implication of ROS generation in the neurotoxicity induced by EPX, indicating that the production of ROS is the main cause of the induction of apoptosis probably via the mitochondrial pathway.
    Keywords:  Apoptosis; Cell cycle arrest; Cytoskeleton; Epoxiconazole; F98 cells; Mitochondria; ROS
    DOI:  https://doi.org/10.1016/j.chemosphere.2019.05.018
  51. Aging (Albany NY). 2019 May 15.
    Zhou G, Hu RK, Xia GC, Yan SH, Ren QL, Zhao J, Wang FH, Huang CC, Yao Q, Tan Y, Zhao NW.
      Many infertile women suffered from poor ovarian response, and increased reactive oxygen species with age might mediate the poor ovarian response to FSH. In this study, we collected follicular fluids and isolated granulosa cells from female patients. Increased levels of peroxynitrite, tyrosine nitrations of FSH receptor (FSHR) and apoptosis were obviously detectable with decreased FSHR protein expressions in granulosa cells of the poor ovarian responders. In KGN (a human ovarian granulosa cell line) cells, exogenous peroxynitrite could sequester FSHR in the cytoplasm, and these dislocated FSHR might suffer from proteasome-mediated degradations. Here, we identified four peroxynitrite-mediated nitrated tyrosine residues of FSHR. Site-directed mutagenesis of FSHR revealed that Y626 was pivotal for intracellular trafficking of FSHR to the cell surface. Akt-induced inactivation of FoxO3a was required for the repression of FSH on granulosa cell apoptosis. However, peroxynitrite impaired FSH-induced Akt-FoxO3a signaling, while FSHR-Y626A mutant took similar effects. In addition, FoxO3a knockdown indeed impaired FSH-mediated cell survival, while FoxO3a-S253A mutant reversed that significantly.
    Keywords:  FSHR; granulosa cells; peroxynitrite; poor ovarian response; tyrosine nitrations
    DOI:  https://doi.org/10.18632/aging.101964
  52. Am J Chin Med. 2019 May 16. 1-23
    Tsai YH, Lin JJ, Ma YS, Peng SF, Huang AC, Huang YP, Fan MJ, Lien JC, Chung JG.
      Fisetin, a naturally occurring flavonoid, is found in common fruits and vegetables and has been shown to induce cytotoxic effects in many human cancer cell lines. No information has shown that fisetin induced cell cycle arrest and apoptosis in mouse leukemia WEHI-3 cells. We found that fisetin decreased total viable cells through G0/G1 phase arrest and induced sub-G1 phase (apoptosis). We have confirmed fisetin induced cell apoptosis by the formation of DNA fragmentation and induction of apoptotic cell death. Results indicated that fisetin induced intracellular Ca 2+ increase but decreased the ROS production and the levels of ΔΨ m in WEHI-3 cells. Fisetin increased the activities of caspase-3, -8 and -9. Cells were pre-treated with inhibitors of caspase-3, -8 and -9 and then treated with fisetin and results showed increased viable cell number when compared to fisetin treated only. Fisetin reduced expressions of cdc25a but increased p-p53, Chk1, p21 and p27 that may lead to G0/G1 phase arrest. Fisetin inhibited anti-apoptotic protein Bcl-2 and Bcl-xL and increased pro-apoptotic protein Bax and Bak. Furthermore, fisetin increased the protein expression of cytochrome c and AIF. Fisetin decreased cell number through G0/G1 phase arrest via the inhibition of cdc25c and induction of apoptosis through caspase-dependent and mitochondria-dependent pathways. Therefore, fisetin may be useful as a potential therapeutic agent for leukemia.
    Keywords:  Apoptosis; Cell Cycle; Fisetin; G/G Phase Arrest; Leukemia; WEHI-3 Cells
    DOI:  https://doi.org/10.1142/S0192415X19500447
  53. Iran J Pharm Res. 2019 ;18(1): 369-382
    Erfani Majd N, Azizian H, Tabandeh MR, Shahriari A.
      Okra (A.esculentus) is an antidiabetic plant whose beneficial effect on ovarian dysfunction in diabetes condition has not been clarified. The present study was designed to examine the effect of Okra powder on serum oxidant/antioxidant status, ovarian structure, and expression of apoptotic/antiapoptotic related genes in ovary of experimentally induced high fat diet diabetic rats. Diabetes was induced by 5 weeks feeding of Wistar rats with high fat diet (HFD) and subsequent i.p injection of STZ (35 mg/kg). Diabetic animals (serum glucose above 250 mg/dL) were treated with Okra powder (200mg/kg) supplemented in diet or metformin (200mg/kg) for 30days. After 30 days of treatment, animals were euthanized and insulin resistance markers (insulin and glucose levels and HOMA-IR), ovarian expression of apoptotic/antiapoptotic genes (Bax, caspase3 and Bcl2) and serum oxidant/antioxidant levels (SOD, GPX and CAT activities and MDA level) were determined. The ovaries were also processed for histological study. Hyperglycemia and reduced body weight of diabetic rats were improved after administration of Okra for 30days. This effect was relatively similar to metformin. Okra resulted in reduction of follicular atresia in concomitant with down regulation of apoptotic related genes (Bax and caspase3) in ovary of diabetic rats. Okra could also diminished oxidative stress in diabetic rats by increasing of serum GPX and CAT activities and reducing the lipid peroxidation level. The results of the present study revealed that Okra powder could be useful intervention for improvement of ovaian dysfunction in diabetic rat by three probable mechanisms; attenuation of glucotoxicity, down regulation of ovarian apoptosis related genes and reduction of oxidative stress.
    Keywords:  Abelmoschus esculentus; Apoptosis; Diabetes; High Fat Diet; Histology; Ovary; Oxidative stress
  54. PLoS One. 2019 ;14(5): e0216604
    K B A, Madhavan A, T R R, Thomas S, Nisha P.
      In this study, the prebiotic potential of soluble dietary fibre extracted from plantain inflorescence (PIF) was investigated. PIF demonstrated prebiotic potential by enhancing the growth of the probiotics under study and thereby hindered colon cancer development. The soluble dietary fibre from Musa paradisiaca inflorescence (PIF) was fermented using Lactobacillus casei and Bifidobacterium bifidum. The fermentation supernatants (LS and BS) were enriched with short chain fatty acids (SCFA) and were able to initiate apoptotic signalling in HT29 colon cancer cells leading to cell death. Both BS and LS exhibited cytotoxic effect; induced DNA damage and enhanced generation of reactive oxygen species in HT29 cells leading to apoptosis. The induction of apoptosis was facilitated by the reduction of membrane potential of mitochondria and ATP synthesis; enhanced delivery of cytochrome c and interference with the expression of pro/antiapoptotic proteins. BS, which exhibited better activity, was further analysed for the identification of differentially regulated proteins by performing two dimensional electrophoresis and MALDI-TOF mass spectrometry. Results emphasized on the fact that, the exposure to BSalteredthe HT29 proteins expression, particularly the upregulation of apoptosis- inducing factor-AIFM1 leading to apoptosis of HT29 cells.
    DOI:  https://doi.org/10.1371/journal.pone.0216604
  55. Photochem Photobiol Sci. 2019 May 17.
    Qi F, Sun Y, Lv M, Qin F, Cao W, Bi L.
      Oral squamous cell carcinoma (OSCC) is a common malignant tumor, accounting for about 7% of all malignant tumors. Palmatine hydrochloride (PaH) is the alkaloid constituent of Fibraurea tinctoria Lour. The present study aims to investigate the antitumor effect of photodynamic therapy (PDT) with PaH (PaH-PDT) on human OSCC cell lines both in vitro and in vivo. The results indicate that PaH-PDT exhibited a potent phototoxic effect in cell proliferation and produced cell apoptosis. PaH-PDT increased the percentage of cells in the G0/G1 phase and decreased the CDK2 and Cyclin E1 protein level. In addition, PaH-PDT markedly increased the generation of intracellular ROS, which can be suppressed using the ROS scavenger N-acetylcysteine (NAC). Furthermore, PaH-PDT increased the expression of p53 protein in vitro and in vivo. In vivo experiments revealed that the PaH-PDT resulted in an effective inhibition of tumor growth and prolonged the survival time of tumor-bearing mice. Moreover, no obvious signs of side effects or a drop in body weight was observed. These results suggested that PaH was a promising sensitizer that can be combined with light to produce significant anti-tumor effects in oral squamous cell carcinoma via enhanced ROS production and up-regulated expression of p53.
    DOI:  https://doi.org/10.1039/c9pp00040b
  56. J Inorg Biochem. 2019 May 04. pii: S0162-0134(19)30102-3. [Epub ahead of print]197 110703
    Liu Z, Li J, Ge X, Zhang S, Xu Z, Gao W.
      A range of phosphorescent Ir(III) complexes containing four diverse P^P-chelating ligands of the type [Ir(ppy)2(L)][PF6], (ppy = 2‑phenylpyridine) where L is 1,2‑bis(diphenylphosphino)benzene (L1), 1,2‑bis(diphenylphosphino)ethane (L2), 1,2‑bis(diphenylphosphino)propane(L3) and 1,8‑bis(diphenylphosphino)naphthalene (L4) were synthesized respectively. The iridium complexes possessed excellent antiproliferative properties, which was a substantial improvement over cisplatin, especially complex Ir1. Generally, the order of in vitro antiproliferative activity of the complexes is Ir1 > Ir2 = Ir3 > Ir4 > CDDP (Cisplatin). Two X-ray crystal structures were determined. The best complex, Ir1, was chosen to further study the mechanism of action. The self-luminescence of complex Ir1 was also successfully used to elucidate the subcellular localization. Complex Ir1 was specifically targeted to lysosomes in A549 cancer cells. This targeting caused lysosomal damage and the induction of ROS (reactive oxygen species) production in cancer cells. Flow cytometry studies confirmed that this complex induced apoptosis, especially late apoptosis. Our results suggested that changes in the mitochondrial membrane potential were responsible for apoptosis. The chemistry and biological studies showed that this class of metal complexes is worthy of further exploration to design novel anticancer drugs.
    DOI:  https://doi.org/10.1016/j.jinorgbio.2019.110703
  57. iScience. 2019 Apr 26. pii: S2589-0042(19)30124-5. [Epub ahead of print]15 196-210
    Porter EG, Dhiman A, Chowdhury B, Carter BC, Lin H, Stewart JC, Kazemian M, Wendt MK, Dykhuizen EC.
      Polybromo1 (PBRM1) is a chromatin remodeler subunit highly mutated in cancer, particularly clear cell renal carcinoma. PBRM1 is a member of the SWI/SNF subcomplex, PBAF (PBRM1-Brg1/Brm-associated factors), and is characterized by six tandem bromodomains. Here we establish a role for PBRM1 in epithelial cell maintenance through the expression of genes involved in cell adhesion, metabolism, stress response, and apoptosis. In support of a general role for PBRM1 in stress response and apoptosis, we observe that loss of PBRM1 results in an increase in reactive oxygen species generation and a decrease in cellular viability under stress conditions. We find that loss of PBRM1 promotes cell growth under favorable conditions but is required for cell survival under conditions of cellular stress.
    Keywords:  Cancer; Molecular Mechanism of Behavior; Molecular Mechanism of Gene Regulation
    DOI:  https://doi.org/10.1016/j.isci.2019.04.027