bims-aporos Biomed News
on Apoptosis and reactive oxygen species
Issue of 2019‒04‒21
seventy-one papers selected by
Gavin McStay
Staffordshire University


  1. Oxid Med Cell Longev. 2019 ;2019 7415212
    Zhao YS, An JR, Yang S, Guan P, Yu FY, Li W, Li JR, Guo Y, Sun ZM, Ji ES.
      Obstructive sleep apnea (OSA) can cause intermittent changes in blood oxygen saturation, resulting in the generation of many reactive oxygen species (ROS). To discover new antioxidants and clarify the endoplasmic reticulum (ER) stress involved in cardiac injury in OSA, we established a chronic intermittent hypoxia (CIH) rat model with a fraction of inspired O2 (FiO2) ranging from 21% to 9%, 20 times/h for 8 h/day, and the rats were treated with H2-O2 mixture (67% hydrogen and 33% oxygen) for 2 h/day for 35 days. Our results showed that H2-O2 mixture remarkably improved cardiac dysfunction and myocardial fibrosis. We found that H2-O2 mixture inhalation declined ER stress-induced apoptosis via three major response pathways: PERK-eIF2α-ATF4, IRE 1-XBP1, and ATF 6. Furthermore, we revealed that H2-O2 mixture blocked c-Jun N-terminal kinase- (JNK-) MAPK activation, increased the ratio of Bcl-2/Bax, and inhibited caspase 3 cleavage to protect against CIH-induced cardiac apoptosis. In addition, H2-O2 mixture considerably decreased ROS levels via upregulating superoxide dismutase (SOD) and glutathione (GSH) as well as downregulating NADPH oxidase (NOX 2) expression in the hearts of CIH rats. All the results demonstrated that H2-O2 mixture significantly reduced ER stress and apoptosis and that H2 might be an efficient antioxidant against the oxidative stress injury induced by CIH.
    DOI:  https://doi.org/10.1155/2019/7415212
  2. Braz J Med Biol Res. 2019 ;pii: S0100-879X2019000400608. [Epub ahead of print]52(4): e7626
    Zhang YE, Huang GQ, Wu B, Lin XD, Yang WZ, Ke ZY, Liu J.
      Reactive oxygen species (ROS) are highly reactive chemical species that may cause irreversible tissue damage, and play a critical role in cardiovascular diseases. Hydrogen sulfide (H2S) is a gasotransmitter that acts as a ROS scavenger with cardio-protective effects. In this study, we investigated the cytoprotective effect of H2S against H2O2-induced apoptosis in cardiomyocytes. H9c2 rat cardiomyoblasts were treated with H2S (100 μM) 24 h before challenging with H2O2 (100 μM). Apoptosis was then assessed by annexin V and PI, and mitochondrial membrane potential was measured using a fluorescent probe, JC-1. Our results revealed that H2S improved cell viability, reduced the apoptotic rate, and preserved mitochondrial membrane potential. An increased Bcl-2 to Bax ratio was also seen in myocytes treated with H2S after H2O2-induced stress. Our findings indicated a therapeutic potential for H2S in preventing myocyte death following ischemia/reperfusion.
    DOI:  https://doi.org/10.1590/1414-431X20187626
  3. Oxid Med Cell Longev. 2019 ;2019 4528616
    Li X, Fang F, Gao Y, Tang G, Xu W, Wang Y, Kong R, Tuyihong A, Wang Z.
      During radiotherapy, reactive oxygen species- (ROS-) induced apoptosis is one of the main mechanism of radiation. Based on KillerRed which can induce ROS burst in different cell substructures, here we hypothesized that KillerRed targeting mitochondria (mtKR) could induce ROS to enhance apoptosis by radiation. In this study, empty vector, mtKR, and mtmCherry plasmids were successfully constructed, and mitochondrial localization were detected in COS-7 and HeLa cells. After HeLa cells were transfected and irradiated by visible light and X-rays, ROS levels, mitochondrial membrane potential (Δψ m), ATPase activities, adenosine triphosphate (ATP) content, apoptosis, and the expressions of mRNA and protein were measured, respectively. Data demonstrated that the ROS levels significantly increased after light exposure, and adding extra radiation, voltage-dependent anion channel 1 (VDAC1) protein increased in the mitochondria, while Na+-K+ and Ca2+-Mg2+ ATPase activities, ATP content, and Δψ m significantly reduced. Additionally, the cell apoptotic rates dramatically increased, which referred to the increase of cytochrome c (Cyt c), caspase-9, and caspase-3 mRNA expressions, and Cyt c protein was released from the mitochondria into the cytoplasm; caspase-9 and -3 were activated. These results indicated that mtKR can increase the production of ROS, enhance mitochondrial dysfunction, and strengthen apoptosis by radiation via Cyt c/caspase-3 pathway.
    DOI:  https://doi.org/10.1155/2019/4528616
  4. Eur J Pharmacol. 2019 Apr 11. pii: S0014-2999(19)30248-1. [Epub ahead of print]
    Viswanathan A, Sebastianelli G, Brown K, Raunio J, Sipilä V, Yli-Harja O, Candeias NR, Kandhavelu M.
      In the present study, a series of L-valine derived boroxazolidones, previously synthesized and reported to have residual activity in a human epithelial cell line, have been evaluated in vitro for their anti-glioblastoma activity. A boroxazolidone derivative containing 2,4-difluorophenyl moieties (6) was found to have higher cytotoxicity than the standard drug, Temozolomide (TMZ). Compound 6 was found to exhibit dose-dependent growth inhibitory effects with an IC50 of 49 μM and 53 μM for LN229 and SNB19 cells, respectively. Additionally, 6 was assessed for its role in apoptosis, caspase 3/7 activation and oxidative stress in SNB19 and LN229 cells. SNB19 cells treated with 6 showed 45.3% apoptosis in the population, while TMZ had 24.7%. In LN229 cells, the percentage of apoptotic cells treated with compound 6 and TMZ were the same. Both 6 and TMZ induced apoptosis through the activation of caspase 3/7 in SNB19 and LN229 cells. Interestingly, 6 exhibited a higher effectivity in promoting reactive oxygen species production in LN229, while it was 6-fold less in SNB19. Boroxazolidone-treated GBM cell lines increased reactive oxygen species production, suggesting that such species may be interlinked with the observed programmed cell death. Additionally, the treatment of both GBM cell lines with 6 led to G2/M phase arrest. The magnitude of anti-GBM effect of 6 is significantly higher than the known chemotherapeutic agent TMZ. This work further demonstrates the anticancer properties of L-valine derived boroxazolidones, adding another potential derivative to the collection of promising chemotherapeutic agents for GBM treatment.
    Keywords:  Apoptosis; Boroxazolidone; Cell cycle arrest; Cytotoxicity; Glioblastoma
    DOI:  https://doi.org/10.1016/j.ejphar.2019.04.020
  5. Int J Mol Sci. 2019 Apr 15. pii: E1865. [Epub ahead of print]20(8):
    Kim JM, Park SK, Kang JY, Park SB, Yoo SK, Han HJ, Cho KH, Kim JC, Heo HJ.
      The aim of this study was to investigate the availability of seeds, one of the byproducts of green tea, and evaluate the physiological activity of seed oil. The ameliorating effect of green tea seed oil (GTO) was evaluated on H₂O₂-induced PC12 cells and amyloid beta (Aβ)1-42-induced ICR mice. GTO showed improvement of cell viability and reduced reactive oxygen species (ROS) production in H₂O₂-induced PC12 cells by conducting the 2',3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) and 2',7'-dichlorofluorescein diacetate (DCF-DA) analysis. Also, administration of GTO (50 and 100 mg/kg body weight) presented protective effects on behavioral and memory dysfunction by conducting Y-maze, passive avoidance, and Morris water maze tests in Aβ-induced ICR mice. GTO protected the antioxidant system by reducing malondialdehyde (MDA) levels, and by increasing superoxide dismutase (SOD) and reducing glutathione (GSH) contents. It significantly regulated the cholinergic system of acetylcholine (ACh) contents, acetylcholinesterase (AChE) activities, and AChE expression. Also, mitochondrial function was improved through the reduced production of ROS and damage of mitochondrial membrane potential (MMP) by regulating the Aβ-related c-Jun N-terminal kinase (JNK)/protein kinase B (Akt) and Akt/apoptosis pathways. This study suggested that GTO may have an ameliorating effect on cognitive dysfunction and neurotoxicity through various physiological activities.
    Keywords:  green tea seed oil, amyloid β, neuroprotective effect, Aβ-related Akt pathway
    DOI:  https://doi.org/10.3390/ijms20081865
  6. Oxid Med Cell Longev. 2019 ;2019 4619865
    Öztürk Y, Günaydın C, Yalçın F, Nazıroğlu M, Braidy N.
      Numerous studies have reported a strong association between increased production of reactive oxygen species (ROS) and the pathobiology of several diseases, and cancer in particular. Therefore, manipulation of cellular oxidative stress levels represents an important therapeutic target. Recently, resveratrol (RESV), a naturally occurring phytochemical, has been shown to sensitize several cell lines to the anticancer effects of other chemotherapeutic agents, including paclitaxel (PAX). However, the molecular mechanisms of action of RESV through oxidative sensitive TRPM2 channel activation remain unclear. The aim of this study was to evaluate the effect of combination therapy of RESV and PAX on activation of TRPM2 in DBTRG glioblastoma cells. DBTRG cells were divided into four treatment groups: control, RESV (50 μM), PAX (50 μM), and PAX + RESV for 24 hours. Our data shows that markers for apoptosis, mitochondrial membrane depolarization and mitochondrial function, intracellular steady-state ROS levels, caspase 3 activity, TRPM2 current density, and Ca2+ florescence intensity were significantly increased in DBTRG cells following treatment with PAX and RESV, respectively, although cell viability was also decreased by these treatments. These biochemical markers were further increased to favor the anticancer effects of PAX in DBTRG cells in combination with RESV. The PAX and RESV-mediated increase in current density and Ca2+ florescence intensity was decreased with a TRPM2 blocker. This suggests that for this combination therapy to have a substantial effect on apoptosis and cell viability, the TRPM2 channel must be stimulated.
    DOI:  https://doi.org/10.1155/2019/4619865
  7. Adv Clin Exp Med. 2019 Apr 15.
    Tanrıkulu-Küçük S, Başaran-Küçükgergin C, Söğüt İ, Tunçdemir M, Doğru-Abbasoğlu S, Seyithanoğlu M, Koçak H, Öner-İyidoğan Y.
      BACKGROUND: Apoptosis plays a major role in fatty liver disease. High-fat diets are related to the onset of fatty liver disease and hepatic oxidant-antioxidant imbalance. Curcumin and capsaicin are somewhat beneficial in reducing hepatic triglycerides; this is most likely because they are known to downregulate reactive oxygen species (ROS) and apoptosis.OBJECTIVES: The aim of this study was to investigate the effects of curcumin and capsaicin on apoptosis through the oxidative effect in an animal model of fatty liver disease.
    MATERIAL AND METHODS: Male Sprague-Dawley rats were fed a normal control diet, a high-fat diet (HFD; 60% of total calories from fat), a HFD+curcumin (1.5 g curcumin/kg HFD), a HFD+capsaicin (0.15 g capsaicin/kg HFD), or a HFD+curcumin+capsaicin (1.5 g curcumin and 0.15 g capsaicin/kg HFD). Liver lysate levels of BAX, Bcl-2 and caspase-3 were determined via immunoblotting. Caspase-3 activity was measured with a colorimetric caspase-3 measurement kit. Total antioxidant status (TAS) and total oxidant status (TOS) were assayed using commercial kits. The generation of hepatic ROS was measured with fluorimetry. Fragmentation of DNA was detected using the TUNEL method.
    RESULTS: High-fat diet caused increased expression of BAX and caspase-3, as well as increased TOS and caspase-3 activity, but decreased expression of Bcl-2. HFD+curcumin+capsaicin caused decreased BAX, caspase-3, TOS, and ROS levels as compared to HFD, but increased TAS and Bcl-2. A HFD +curcumin+capsaicin also decreased the number of TUNEL-positive cells.
    CONCLUSIONS: These results suggest that supplementation with curcumin and capsaicin balances the hepatic oxidant-antioxidant status and may have a protective role in the apoptotic process in an HFD-induced fatty liver model.
    Keywords:  apoptosis; capsaicin; curcumin; fatty liver; high-fat diet
    DOI:  https://doi.org/10.17219/acem/94145
  8. Hum Exp Toxicol. 2019 Apr 17. 960327119843578
    Katubi KM, Alzahrani FM, Ali D, Alarifi S.
      Nanotechnology has achieved more commercial attention over recent years, and its application has increased concerns about its discharge in the environment. In this study, we have chosen human hepatic carcinoma (HuH-7) cells because liver tissue has played an important role in human metabolism. Therefore, the objective of this study was to determine DNA damaging and apoptotic potential of cadmium telluride quantum dots (CdTe QDs; average particle size (APS) 10 nm, 1-25 µg/ml) on HuH-7 cells and the basic molecular mechanism of its cellular toxicity. Cytotoxicity of different concentrations of CdTe QDs on HuH-7 cells was determined by 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS) and lactate dehydrogenase (LDH) tests. Moreover, reactive oxygen species (ROS) generation, mitochondrial membrane potential, DNA damage, and Hoechst 33342 fluorescent staining morphological analysis of necrotic/apoptotic cells were detected; cellular impairment in mitochondria and DNA was confirmed by JC-1 and comet assay, respectively. A dose- and time-dependent cytotoxicity effect of CdTe QDs exposure was observed HuH-7 cells; the significant ( p < 0.05) cytotoxicity was found at 25 μg/ml of CdTe QDs exposure. The percentage of cytotoxicity of CdTe QDs (25 μg/ml) in HuH-7 cells reached 62% in 48 h. CdTe QDs elicited intracellular ROS generation and mitochondrial depolarization, and DNA integrity cells collectively advocated the apoptotic cell death at higher concentration. DNA damage was observed in cells due to CdTe QDs exposure, which was mediated by oxidative stress. This study exploring the effects of CdTe QDs in HuH-7 cells has provided valuable insights into the mechanism of toxicity induced by CdTe QDs.
    Keywords:  CdTe QDs; DNA damage; HuH-7 cells; ROS; apoptosis; comet assay
    DOI:  https://doi.org/10.1177/0960327119843578
  9. Mol Reprod Dev. 2019 Apr 15.
    Sun J, Yin B, Tang S, Zhang X, Xu J, Bao E.
      Heat stress is a major stressor that can lead to male reproductive dysfunction. Sertoli cells play a crucial role in spermatogenesis by providing germ cells with structural and nutritional support, and contributing to blood-testis barrier formation. Vitamin C (Vc) is an antioxidant capable of neutralizing reactive oxygen species and preventing lipid peroxidation widely used because it is inexpensive and highly accessible. In the present study, we investigated the protective effect of Vc on TM4 cells following heat stress. Pretreatment with Vc could effectively inhibit apoptosis (p < 0.01), lipid peroxidation, and lactate dehydrogenase (LDH) activity. However, a significant increase in the malondialdehyde (MDA) level and LDH activity (p < 0.01) was observed in TM4 cells without Vc-pretreatment, in conjunction with vacuole degeneration and karyopyknosis. In addition, both the messenger RNA and protein levels of CryAB, Hsp27, Hsp70, and Hsp110 substantially increased in the 3 and 12 hr recovery groups (p < 0.01). Vc also prevented microtubule aggregation following heat stress. These results suggest that pretreatment with Vc-protected TM4 cells against heat stress by reducing the level of oxidative stress and inducing heat shock protein expression.
    Keywords:  TM4 cells; heat shock proteins; heat stress; oxidative damage; vitamin C
    DOI:  https://doi.org/10.1002/mrd.23146
  10. Oxid Med Cell Longev. 2019 ;2019 1292891
    Yang Y, Pang B, Liu Z, Li J, Zhang Z, Zhang R, Hou X, Guo H, Chi L, Pang Q, Xin T.
      Several in vitro studies have revealed the neurotoxicity of 1-trichloromethyl-1,2,3,4-tetrahydro-beta-carboline (TaClo). However, the underlying mechanism has not been completely elucidated, particularly in vivo. This study was designed to study the neurotoxicity of TaClo in vivo by stereotactically injecting TaClo into the striatum of Wistar rats. After the TaClo injections, rats were subjected to an open field test, and their distance travelled and tracks showed decreasing trends over time. The results of liquid chromatography-mass spectrometry analysis showed that the motor dysfunction of the TaClo-treated rats was accompanied by reduced dopamine levels in the striatum. Based on the diffusion tensor imaging data, the apparent diffusion coefficient of the nigrostriatal pathway was significantly increased, and subsequent histological staining revealed the demyelination of nigrostriatal fibres after the TaClo treatment. TaClo induced a loss of tyrosine hydroxylase-positive cells in the substantia nigra compacta. Regarding the underlying mechanism, TaClo caused oxidative stress in the nigrostriatal system by increasing the production of reactive oxygen species and reducing the mitochondria membrane potential. Meanwhile, the elevated expression of Iba-1, TNF-α, IL-6, Cox-2, and iNOS indicated microglial activation and a strong innate immune response in the nigrostriatal system. In addition, activated caspase-3 levels were increased. Thus, both mitochondrial impairments and the innate immune response are involved in TaClo-induced neurotoxicity.
    DOI:  https://doi.org/10.1155/2019/1292891
  11. J Dermatol Sci. 2019 Apr 02. pii: S0923-1811(19)30072-6. [Epub ahead of print]
    Jiang W, Li S, Chen X, Zhang W, Chang Y, He Y, Zhang S, Su X, Gao T, Li C, Jian Z.
      BACKGROUND: Oxidative stress plays important roles in the pathogenesis of vitiligo. The removal of hydrogen peroxided (H2O2) has been established to be beneficial to vitiligo patients. Berberine (BBR), a natural isoquinoline alkaloid, has antioxidant activity, however, whether BBR can defend human melanocytes against oxidative injury remains to be elucidated.OBJECTIVE: In the present study, we investigated the potential protective effect of BBR against oxidative stress on an immortalized normal human melanocyte cell line PIG1.
    METHODS: Generally, PIG1 cells were pretreated with various concentrations of BBR for 1 h followed by exposure to 1.0 mM H2O2 for 24 h. Cell apoptosis, intracellular reactive oxygen species (ROS) levels were assessed through flow cytometry. Cell apoptosis, melanogenesis and the activation of Nrf2-ARE and Mitf signaling pathway were assayed.
    RESULTS: Our results showed that cell viability rose and intracellular ROS generation, cell apoptosis of melanocytes decreased significantly in response to H2O2 through pretreatment with BBR. Furthermore, We found that BBR can dramatically induce Nrf2 nuclear translocation, increase total Nrf2 levels and enhance ARE activity. Besides, Nrf2-siRNA transfection can abrogate the protection of BBR in melanocytes against oxidative injury. At last, we verified that BBR could facilitate melanogenesis function via modulation of Mitf and its target proteins.
    CONCLUSION: The results above suggest that BBR can protect melanocytes against oxidative stress via its anti-oxidative activity. Also, we found H2O2-induced activation of NFκB was inhibited by BBR. Therefore, it is worthy of investigation BBR as a potential drug for treatment of vitiligo.
    Keywords:  Berberine; Melanocyte; Mitf; Nrf2; Oxidative stress
    DOI:  https://doi.org/10.1016/j.jdermsci.2019.03.007
  12. Biomed Pharmacother. 2019 Apr 15. pii: S0753-3322(18)38675-X. [Epub ahead of print]114 108866
    Meng N, Mu X, Lv X, Wang L, Li N, Gong Y.
      Fascaplysin is a natural product isolated from marine sponges that exhibits broad anticancer activity. Previous studies revealed that fascaplysin-induced apoptosis and angiogenesis inhibition in vascular endothelial cells contributed to its anticancer activity. Accumulating evidence indicates that autophagy plays a significant role in mediating the function of vascular endothelial cells (VECs) and the response to cancer therapy. However, the effect of fascaplysin on VEC autophagy and the role of autophagy in fascaplysin-induced vascular endothelial cell apoptosis and angiogenesis inhibition are not clear. The present study found that fascaplysin induced autophagy in vascular endothelial cells. Suppression of autophagy using a pharmacological inhibitor (3-methyladenine) or RNA interference of an essential autophagy gene (ATG5) enhanced the cell death and anti-angiogenesis activity of fascaplysin. We further found that fascaplysin significantly increased p8 protein and reactive oxygen species (ROS) levels and decreased mitochondrial membrane potential but had no effect on the mTOR pathway in VECs. Notably, the ROS scavenger N-acetylcysteine inhibited fascaplysin-induced autophagy and increased p8 protein level. Knockdown of p8 by using RNA interference inhibited the autophagy but increased the level of ROS in VECs. Taken together, these data indicated that fascaplysin activated autophagy as a cytoprotective response via ROS and p8 in VECs. Our findings provided important insight into the response of VECs to fascaplysin and may be useful for improving the anticancer efficacy of fascaplysin.
    Keywords:  Angiogenesis; Apoptosis; Autophagy; Fascaplysin; Vascular endothelial cells
    DOI:  https://doi.org/10.1016/j.biopha.2019.108866
  13. Environ Pollut. 2019 Apr 04. pii: S0269-7491(19)30153-8. [Epub ahead of print]250 58-67
    Yuan S, Han Y, Ma M, Rao K, Wang Z, Yang R, Liu Y, Zhou X.
      Aryl phosphorus-containing flame retardants (aryl-PFRs) have been frequently detected with increasingly used worldwide as one of alternatives for brominated flame retardants. However, information on their adverse effects on human health and ecosystem is insufficient, with limited study on their molecular mode of action in vitro. In this study, the cytotoxicity, DNA damage, mitochondrial impairment and the involved molecular mechanisms of certain frequently detectable aryl-PFRs, including 2-ethylhexyldiphenyl phosphate (EHDPP), methyl diphenyl phosphate (MDPP), bisphenol-A bis (diphenyl phosphate) (BDP), isodecyl diphenyl phosphate (IDPP), cresyl diphenyl phosphate (CDP) and the structurally similar and widely used organophosphorus pesticide chlorpyrifos (CPF), were evaluated in A549 cells using high-content screening (HCS) system. Aryl-PFRs showed different lethal concentration 50 (LC50) values ranging from 97.94 to 546.85 μM in A549 cells using CCK-8 assay. EHDPP, IDPP, CDP, MDPP and CPF demonstrated an ability to induce DNA damage, evidenced by increased DNA content and S phase-reducing cell cycle arrest effect using fluorophore dye cocktail assay. Additionally, the selected aryl-PFRs induced mitochondrial impairment by the increasing mitochondrial mass and decreasing mitochondrial membrane potential. Moreover, BDP, MDPP, and CDP, which contain short alkyl chains showed their potential oxidative stress with intracellular ROS and mitochondrial superoxide overproduction from an initially relatively low concentration. Additionally, based on the promotion of firefly luminescence in p53-transfected A549 cells, p53 activation was found to be involved in aryl-PFRs-induced DNA damage. Further real-time PCR results showed that all selected aryl-PFRs triggered p53/p21/gadd45β-, and p53/p21/mdm2-mediated cell cycle pathways, and the p53/bax mediated apoptosis pathway to induce DNA damage and cytotoxic effects. These results suggest that aryl-PFRs (e.g., BDP, MDPP, CDP) cause oxidative stress-mediated DNA damage and mitochondrial impairment, and p53-dependent pathway was involved in the aryl-PFRs-induced DNA damage and cell cycle arrest. In conclusion, this study improves the understanding of PFRs-induced adverse outcomes and the involved molecular mechanism.
    Keywords:  Gene expression; Mitochondrial damage; Phosphorus-containing flame retardants; Reactive oxygen species; p53 transfection
    DOI:  https://doi.org/10.1016/j.envpol.2019.03.109
  14. Exp Ther Med. 2019 May;17(5): 4213-4222
    Hsiao CH, Yao CJ, Lai GM, Lee LM, Whang-Peng J, Shih PH.
      Bladder cancer is one of the most common types of malignant tumor worldwide. Current treatments, including chemo-/radiotherapy, only have limited efficacy on bladder cancer progression. Honokiol is an active component of Magnolia officinalis with multiple biological effects that may provide promising health benefits. In the present study, the anti-cancer properties of honokiol against bladder cancer cells were investigated by flow cytometric analysis. The results revealed that honokiol exhibited significant anti-proliferative effects on bladder cancer cell lines, particularly on BFTC-905 human transitional cell carcinoma cells. Furthermore, honokiol at low doses (≤25 µM) induced cell cycle arrest in G0/G1 phase, while it induced significant apoptotic cell death at high doses (≥50 µM; P<0.05). Furthermore, a significant accumulation of reactive oxygen species was identified in honokiol-treated cells. In addition, honokiol induced hyperpolarization of the mitochondrial membrane, which may lead to mitochondrial dysfunction. Finally, caspase-3/7 activation was identified in high-dose honokiol-treated bladder cancer cells. These results suggest that honokiol induces apoptosis via the mitochondrial pathway and honokiol-containing traditional herbal remedies may have a potential clinical application in the treatment of bladder cancer.
    Keywords:  apoptosis; bladder cancer; flow cytometry; honokiol; magnolia officinalis; mitochondrial membrane hyperpolarization; oxidative stress
    DOI:  https://doi.org/10.3892/etm.2019.7419
  15. J Neural Transm (Vienna). 2019 Apr 19.
    Jiang DQ, Ma YJ, Wang Y, Lu HX, Mao SH, Zhao SH.
      Microglia activation-mediated neuroinflammation plays an important role in the progression of Parkinson's disease (PD). However, effects of microglia activation on dopaminergic neuronal cell (DAC) fate are still poorly understood. The objective of this study was to explore the neurotoxic effects of microglia activation-mediated oxidative injury in DACs and its possible mechanisms. In the present study, microglia-DACs co-culture systems (murine BV-2 and MN9D cells, or primary microglia and mesencephalic neurons) were used to display the crosstalk between both cell types. The cytotoxicity of lipopolysaccharide-induced microglia activation led to the accumulation of intracellular reactive oxygen species, increased cell apoptosis rate, reduced number of DACs, concomitant to cell cycle arrest at G1 phase. Molecular mechanisms of apoptosis caused by microglia activation-induced oxidative injury included the increased opening of mitochondrial permeability transition pore and enhanced membrane potential depolarization in MN9D cells, down-regulation of Bcl-2 and up-regulation of Bax, caspase-3 expression in DACs. In addition, microglia activation made a significant reduction of SIRT3 and superoxide dismutase 2 gene expression in DACs. Taken together, these data imply that microglia activation promotes cell apoptosis through mitochondrial pathway and decreases SIRT3 expression in DACs, which may provide some support for PD progression promoted by neuroinflammation.
    Keywords:  Apoptosis; Dopaminergic cell; Microglia activation; Oxidative injury; SIRT3
    DOI:  https://doi.org/10.1007/s00702-019-02005-z
  16. Atherosclerosis. 2019 Apr 10. pii: S0021-9150(19)30357-0. [Epub ahead of print]285 23-30
    Yang S, Zhang W, Xuan LL, Han FF, Lv YL, Wan ZR, Liu H, Ren LL, Gong LL, Liu LH.
      BACKGROUND AND AIMS: Akebia Saponin D (ASD) is a major bioactive triterpenoid saponin compound isolated from the Chinese herb Dipsacus asper wall (DSW). DSW has been long used as an anti-Alzheimer disease and anti-osteoporosis agent in clinics. However, anti-atherosclerotic effects of ASD have not been fully investigated. The objective of this study is to further investigate the anti-atherosclerotic activities and mechanisms of ASD in vivo and in vitro.METHODS: In in vitro experiments, ASD (50, 100, and 200 μM) was used to explore the effects of preventing H2O2-induced endothelial cell apoptosis and the possible mechanism involved. In in vivo experiments, ApoE-/- mice were fed a high fat diet (HFD) and treated with atorvastatin (10 mg/kg/d), ASD (50, 150, 450 mg/kg/d), or the combination therapy (atorvastatin 10 mg/kg/d and ASD 150 mg/kg/d) for 14 weeks.
    RESULTS: We found that ASD reduced the generation of reactive oxygen species, inhibited mitochondrial membrane potential (MMP) impairment, diminished the expression of Bax and Caspase-3, increased Bcl-2 expression, and inhibited apoptosis in endothelial cells. ASD significantly increased the expression of anti-oxidant enzymes (GSH, SOD, and CAT) in both liver and vascular tissue, reduced blood lipid levels (TG, TC, and LDL-C), and decreased lipid deposition in the liver and atherosclerotic lesion size in ApoE-/- mice.
    CONCLUSIONS: Our study revealed that ASD inhibited atherosclerosis development in ApoE-/- mice by inhibiting oxidative stress-induced endothelial cell apoptosis signaling pathway, and suggested that ASD might be a potential therapeutic drug in the prevention of atherosclerosis.
    Keywords:  Akebia saponin D; Antioxidant; Apoptosis; Atherosclerosis
    DOI:  https://doi.org/10.1016/j.atherosclerosis.2019.04.202
  17. Front Pharmacol. 2019 ;10 330
    Doan P, Musa A, Candeias NR, Emmert-Streib F, Yli-Harja O, Kandhavelu M.
      Glioblastoma (GBM) is the most common type of malignant brain tumor in adults. We show here that small molecule 2-[(3,4-dihydroquinolin-1(2H)-yl)(p-tolyl)methyl]phenol (THTMP), a potential anticancer agent, increases the human glioblastoma cell death. Its mechanism of action and the interaction of selective signaling pathways remain elusive. Three structurally related phenolic compounds were tested in multiple glioma cell lines in which the potential activity of the compound, THTMP, was further validated and characterized. Upon prolonged exposer to THTMP, all glioma cell lines undergo p53 and cyclin-dependent kinase mediated cell death with the IC50 concentration of 26.5 and 75.4 μM in LN229 and Snb19, respectively. We found that THTMP strongly inhibited cell growth in a dose and in time dependent manner. THTMP treatment led to G1/S cell cycle arrest and apoptosis induction of glioma cell lines. Furthermore, we identified 3,714 genes with significant changes at the transcriptional level in response to THTMP. Further, a transcriptional analysis (RNA-seq) revealed that THTMP targeted the p53 signaling pathway specific genes causing DNA damage and cell cycle arrest at G1/S phase explained by the decrease of cyclin-dependent kinase 1, cyclin A2, cyclin E1 and E2 in glioma cells. Consistently, THTMP induced the apoptosis by regulating the expression of Bcl-2 family genes and reactive oxygen species while it also changed the expression of several anti-apoptotic genes. These observations suggest that THTMP exerts proliferation activity inhibition and pro-apoptosis effects in glioma through affecting cell cycle arrest and intrinsic apoptosis signaling. Importantly, THTMP has more potential at inhibiting GBM cell proliferation compared to TMZ, the current chemotherapy treatment administered to GBM patients; thus, we propose that THTMP may be an alternative therapeutic option for glioblastoma.
    Keywords:  anticancer; apoptosis induction; cell cycle; cytotoxicity; gene expression; phenol
    DOI:  https://doi.org/10.3389/fphar.2019.00330
  18. Life Sci. 2019 Apr 12. pii: S0024-3205(19)30270-X. [Epub ahead of print]
    Hassan E, Kahilo K, Kamal T, El-Neweshy M, Hassan M.
      AIMS: The present study aimed to investigate the potential therapeutic effect of diallyl sulfide (DAS), a natural component of garlic (Allium sativum), in the improvement of lead (Pb)-induced testicular toxicity and its underlying mechanisms.MATERIALS AND METHODS: Thirty-two male albino rats were randomly divided into control, PbAc (20 mg lead acetate/kg bwt, orally), DAS (200 mg/kg bwt, orally), and PbAc + DAS groups for 49 successive days. The investigation based on the following criteria: Paired testes and epididymides weights, epididymal sperm analysis, level of serum sex hormones (Testosterone and17β-estradiol (E2)), aromatase (CYP19) expression, Malondialdehyde (MDA), Nitric oxide (NO), Superoxide dismutase (SOD) enzyme, reduced glutathione (GSH), testicular histopathology, spermatogenesis score and apoptosis detection (Caspase-3 immunoexpression).
    KEY FINDINGS: Pb caused significant decline in epididymal sperm count and motility, testes and epididymes weights, spermatogenesis score and serum testosterone and1E2, as well as a significant decrease in SOD and GSH level, and a significant elevation of MDA and NO compared to a control group. In addition, Pb induced significant downregulation of CYP19 gene expression, increase of Caspase-3 immunoreactivity, various testicular degenerative and necrotic changes. Whereas, co-treatment of rats with DAS improved sperm analysis, and testicular histology and antioxidative status. Furthermore, DAS co-administration regulated testicular CYP19 and Caspase-3 expressions.
    SIGNIFICANCE: Collectively, DAS seemed to be a promising agent for protection against Pb-induced testicular toxicity through antioxidative properties, beside regulation of testicular apoptosis and aromatase expression.
    Keywords:  Aromatase P450 gene; Caspase-3; Diallyl sulfide; Lead; Male infertility; Oxidative stress
    DOI:  https://doi.org/10.1016/j.lfs.2019.04.020
  19. Iran J Kidney Dis. 2019 Mar;13(2): 74-86
    Modanloo M, Shokrzadeh M.
      Mitochondrial dysfunction, apoptosis and oxidative stress, are the interrelated events underlining the pathology of  numerous diseases including cardiovascular, neurologic, and metabolic disorders. Due to playing a critical role in glucose and fatty acids' metabolism, L-carnitine probably has the potential to adjust these unfavorable events. The present review has evolved based on existing literature that investigated the mechanisms of L-carnitine and its derivatives based mitochondrial dysfunction, oxidative stress, and apoptosis related modulation. The released studies have been searched with the databases including Google Scholar, Scopus, and PubMed out of which overall 76 full-length articles have been chosen and recruited in this review. L-carnitine exerts protective effects against these cellular events in several manners including the maintenance of mitochondrial functions and decreasing the production of  reactive oxygen species at different points. In clinical setting, these effects could be applied to treat a variety of associated diseases.
  20. Int J Neurosci. 2019 Apr 17. 1-17
    Cao C, Zhang Y, Zhang Z, Chen Q.
      BACKGROUND: Ketamine (KET) is known to be used in pediatric anaesthetic, whereas many evidences revealed that KET generated neurotoxicity. In our study, we found KET decreased the level of LncRNA taurine-upregulated gene 1 (TUG1). Hence, the aim of our study is to investigate whether small interfering TUG1 (siTUG1) has effect on KET-induced rat hippocampal neurons.MATERIAL AND METHODS: siTUG1 and KET alone or in combination was used to treat with neurons for 12 h. MTT assays was used to detect hippocampal neurons viability. Cell apoptosis and reactive oxygen species (ROS) level were analyzed by flow cytometry assay. TUG1 level was determined via reverse transcription-quantitative polymerase chain reaction (RT-qPCR) assay. The levels of Bax, Bcl-2, cleaved-caspase-3, p38 and p-p38 were measured by western blot (WB) assay.
    RESULTS: Neurons treated by siTUG1 and KET had a higher optical density (OD) value,a lower apoptosis rate and lower ROS level. Neurons treated by the combination of siTUG1 and KET had lower levels of TUG1, cleaved-caspase-3 and p-p38 than those under KET treatment. Besides, siTUG1 + KET group (siTUG1 and KET treated neurons) had a higher Bcl-2 level than KET group.
    CONCLUSION: We found that siTUG1 decreased KET-induced hippocampal neurons apoptosis and ROS level, and increased hippocampal neurons viability. siTUG1 decreased neurons aopotosis via signaling pathways of Bax/Bcl-2 and Caspases, and increased neurons viability by signaling pathway of p38 MAPK. Our results indicate that KET-induced neurotoxicity might be reduced by inhibiting TUG1.
    Keywords:  Apoptosis; Ketamine; Rat hippocampal neurons; TUG1; Viability
    DOI:  https://doi.org/10.1080/00207454.2019.1594805
  21. Cell Stress Chaperones. 2019 Apr 15.
    Zhao MX, Wen JL, Wang L, Wang XP, Chen TS.
      Artesunate (ARS) induced significant reactive oxygen species (ROS) generation in HepG2, HeLa, and A549 lines. However, ARS induced ROS-dependent apoptosis in HeLa and A549 cell lines but ROS-independent apoptosis in HepG2 cells. A total of 200 μM hydrogen peroxide (H2O2) significantly induced cytotoxicity in HeLa cells, while H2O2 up to 300 μM did not induce cytotoxicity in HepG2 cells, further demonstrating the strong resistance of HepG2 cells to ROS. HeLa cells had much higher basic total glutathione (T-GSH) level than HepG2 cells, while the ratio of basic reduced glutathione (GSH)/oxidized glutathione (GSSG) in HepG2 cells was nearly twice than that in HeLa and A549 cells. Inhibition of glutathione markedly enhanced H2O2- or ARS-induced cytotoxicity in HeLa and A549 cell lines but modestly enhanced the cytotoxicity of H2O2 and even did not affect the cytotoxicity of ARS in HepG2 cells. Moreover, addition of GSH remarkably prevented H2O2- or ARS-induced cytotoxicity in HeLa and A549 cell lines, further indicating the involvement of GSH in scavenging ROS in the two cell lines. HepG2 cells exhibited higher catalase activity than HeLa cells, and inhibiting catalase activity by using 3-aminotriazole (3-AT, a specific inhibition of catalase) or catalase siRNA remarkably reduced the resistance of HepG2 cells to ROS, demonstrating the key roles of catalase for the strong resistance of HepG2 cells to ROS. Collectively, catalase activity instead of glutathione level dominates the resistance of cells to ROS.
    Keywords:  Catalase; Glutathione; Hydrogen peroxide; Reactive oxygen species; Resistance
    DOI:  https://doi.org/10.1007/s12192-019-00993-1
  22. Life Sci. 2019 Apr 16. pii: S0024-3205(19)30286-3. [Epub ahead of print]
    Handali S, Moghimipour E, Kouchak M, Ramezani Z, Amini M, Angali KA, Saremy S, Dorkoosh FA, Rezaei M.
      We previously showed that folate liposomes of 5FU made from Dipalmitoylphosphatidylcholine (DPPC) induced cell death in HT-29 and HeLa cells more potently than bulk 5FU. Also, a primary 5FU liposomal formulation with phosphatidyl choline (PC) exhibited higher cytotoxicity in murine colon cancer cells. In the present study, optimization of 5FU PC liposome, mechanism of cell death induction in human cancer cell lines and its safety along with other assays have been employed for targeted PC liposomes of 5FU. Liposomes were prepared using thin layer method and optimization of preparation was assessed using central composite design (CCD) of response surface methodology (RSM). Folic acid (FA) was employed as the targeting ligand. Morphology of 5FU loaded liposomes and changes in their thermal behavior were assessed by transmission electron microscopy (TEM) and differential scanning calorimetry (DSC), respectively. In vitro cytotoxicity was explored using MTT assay in HT-29, Caco-2, HeLa and MCF-7 cell lines. Cytotoxicity mechanism of the targeted delivery system was searched through the evaluation of reactive oxygen species (ROS) overproduction, mitochondrial membrane potential (∆Ψm), the release of cytochrome c, the activity of caspase 3/7 and apoptosis and necrosis rate. Liposomes were spherical in shape and 5FU was successfully encapsulated into liposomes rather in an amorphous state. Our interesting results showed that in HT-29 cells targeted liposomes triggered the mitochondrial apoptotic pathway by decreasing the mitochondrial membrane potential, releasing of cytochrome c and promoting the substantial activity of caspase 3/7. In HeLa cells, however, targeted liposomes particularly activated necrosis pathway through the overproduction of ROS. Folate-liposomal 5FU showed significantly higher antitumor efficiency compared to free drug. The results of this study offer new prospects for cancer therapy with reducing systemic drug exposure and associated toxicities.
    Keywords:  5-Fluorouracil; Folic acid; Liposome; Response surface methodology; Targeted drug delivery
    DOI:  https://doi.org/10.1016/j.lfs.2019.04.030
  23. Curr Mol Med. 2019 Apr 15.
    Yang J, Zhu D, Wen L, Xiang X, Hu J.
      OBJECTIVE: To investigate the protective effects of Gentianella turkestanerum extraction by butanol (designated as GBA) on hepatic cell line L02 injury induced by carbon tetrachloride (CCl4) and hydrogen peroxide (H2O2).METHODS: L02 cells were incubated with 5 μg/mL, 10 μg/mL, 20 μg/mL, 40 μg/mL, 60 μg/mL, 80 μg/mL and 100 μg/mL GBA for 24 hours, and then MTT assay was used to screen the cytotoxicity for GBA. Cells were divided into blank control group, CCl4/H2O2 model group, treated by CCl4 (20 mmol/L) or H2O2 (100 μmol/L); silymarin+CCl4/H2O2 group, treated by CCl4 (20 mmol/L) or H2O2 (100 μmol/L) and 5 μg/mL silymarin; GBA+CCl4/H2O2 group, treated by CCl4 (20 mmol/L) or H2O2 (100 μmol/L) and GBA (5 μg/mL, 10 μg/mL and 20 μg/mL). MTT assay was performed to determine the cellular activity. Malondialdehyde (MDA) content was determined using commercial kit. The alanine transaminase (ALT), aspartate transaminase (AST) in the supernatant was determined. PE-Annexin V/7-ADD method was utilized to determine the apoptosis of cells. RT-PCR was used to evaluate the expression of endoplasmic reticulum stress related genes (CHOP, PERK, IRE1 and ATF6) mRNA. Western blot analysis was performed to determine the expression of CHOP, Caspase 12 and NF-κB protein.
    RESULTS: Cellular survival after GBA (5 μg/mL, 10 μg/mL and 20 μg/mL) incubation was ≥ 75%. After GBA incubation, levels of ALT and AST showed significant decrease (P < 0.05), while that of the MDA showed significant decrease (P < 0.05). The apoptosis in the CCl4 or H2O2 group showed significant increase compared to the control group (P < 0.05). In contrast, GBA-preincubation could attenuate the cellular apoptosis compared to the CCl4 or H2O2 group, which displayed a dose-dependent manner (P < 0.05). The expression of CHOP, PERK, IRE1 and ATF6 mRNA was significantly up-regulated in the presence of CCl4 or H2O2 (P < 0.05). Whereas, GBA induced significant decrease of these mRNA there after (P < 0.05), together with decrease in CHOP and Caspase 12 proteins (P < 0.05). Besides, it could attenuate the expression of NF-κB p65 in nuclear protein.
    CONCLUSIONS: G. turkestanerum could inhibit the lipid peroxidation and increase the anti-oxidant activity. Also, it could inhibit the cellular apoptosis through down-regulating the transcriptional level of ERS related genes and proteins. This process was associated with the nuclear translocation of NF-κB p65 protein.
    Keywords:  Gentianella turkestanerum; Hepatic L02 cell; butanol ; carbon tetrachloride; hydrogen peroxide; liver injury
    DOI:  https://doi.org/10.2174/1566524019666190415124838
  24. Oxid Med Cell Longev. 2019 ;2019 2691514
    Zhang B, Zhang G, Wei T, Yang Z, Tan W, Mo Z, Liu J, Li D, Wei Y, Zhang L, Webster KA, Wei J.
      Background and Aims: Vascular smooth muscle cells (VSMCs) are central components of atherosclerotic plaque. Loss of VSMCs through apoptotic cell death can cause fibrous cap thinning, necrotic core formation, and calcification that may destabilize plaque. Elevated glucocorticoid levels caused by psychological stress promote VSMC apoptosis and can exacerbate atherosclerosis in mice and humans. Changes in the levels of antiapoptosis microRNA-25 (miR-25) have been linked with heart disease, inflammation, VSMC phenotype, oxidative stress, and apoptosis. Here, we investigated the pathways and mechanisms of glucocorticoid-induced apoptosis of mouse VSMCs and the protective role of miR-25.Methods: Primary mouse VSMCs were cultured +/- corticosterone for 48 h. Apoptosis, ROS, apoptotic protein activities, miR-25, MOAP1, a miR-25 target, and p70S6 kinase were quantified at intervals. The roles of miR-25 were assessed by treating cells with lenti-pre-miR-25 and anti-miR-25.
    Results: VSMC apoptosis, caspase-3 activity, and Bax were increased by corticosterone, and cell death was paralleled by marked loss of miR-25. Protection was conferred by pre-miR-25 and exacerbated by anti-miR-25. Pre-miR-25 conferred reduced expression of the proapoptotic protein MOAP1, and the protective effects of pre-miR-25 were abrogated by overexpressing MOAP1. The antiapoptotic effects of miR-25 were paralleled by inhibition of the p70S6K pathway, a convergence target for the survival signaling pathways, and protection by pre-miR-25 was abrogated by the p70S6k inhibitor rapamycin.
    Conclusions: MicroRNA-25 blocks corticosterone-induced VSMC apoptosis by targeting MOAP1 and the p70S6k pathway. Therapeutic manipulation of miR-25 may reduce atherosclerosis and unstable plaque formation associated with chronic stress.
    DOI:  https://doi.org/10.1155/2019/2691514
  25. Free Radic Biol Med. 2019 Apr 12. pii: S0891-5849(19)30280-1. [Epub ahead of print]
    Zhang X, Xia Q, Wei R, Song H, Mi J, Lin Z, Yang Y, Sun Z, Zou K.
      Busulfan is a widely used chemotherapeutic drug for chronic myelogenous leukemia and bone marrow transplantation. As a cell cycle nonspecific alkylation agent, busulfan has a severe side effect on germ cells, especially on spermatogonia before meiosis. Studies have revealed that busulfan causes DNA strand crosslinks in spermatogonia and induces apoptosis, and many corresponding strategies have been developed to ameliorate the side effects. However, fertility maintenance after busulfan treatment is still a challenging project in the clinic. Here, we demonstrated that continuous injection of melatonin effectively alleviated germline cytotoxicity both in recipient mice and cultured spermatogonia, and busulfan/melatonin recipient mice produced normal litters. We further revealed that melatonin rescues spermatogonia from apoptosis by neutralizing reactive oxidative species (ROS) induced by busulfan and recovered the phosphorylation of ATM and p53 to normal levels, and as a result apoptosis in spermatogonial progenitor cells was avoided. This study reports that pineal gland hormone melatonin effectively protects spermatogonia from the stress of chemotherapy and oxidation and reveals the underlying molecular mechanisms, which will provide an important hint for fertility protection in clinic.
    Keywords:  ATM; Apoptosis; Busulfan; Fertility; Spermatogonial stem cells; p53
    DOI:  https://doi.org/10.1016/j.freeradbiomed.2019.04.009
  26. Aging (Albany NY). 2019 Apr 19.
    Zhang Y, ShiYang X, Zhang Y, Li Y, Shi X, Xiong B.
      Aristolochic acid (AA) is a class of carcinogenic and nephrotoxic nitrophenanthrene carboxylic acids naturally found in Aristolochia plants. These plants have been widely used as herbal medicines and also enter the human food chain as the persistent soil pollutants. It has been known that AA exposure is implicated in multiple cancer types, kidney failure and ovarian dysfunction. However, whether AA exposure would influence the oocyte quality has not yet determined. Here, we document that AAI has the negative effects on the competency of oocyte maturation and fertilization. We show that AAI exposure leads to the oocyte meiotic failure via impairing the meiotic apparatus, displaying a prominently defective spindle assembly, actin dynamics and mitochondrial integrity. AAI exposure also causes the abnormal distribution of cortical granules and ovastacin, which is consistent with the observation that fewer sperm bound to the zona pellucida surrounding the unfertilized AAI-exposed eggs, contributing to the fertilization failure. In addition, AAI exposure induces the increased levels of ROS, DNA damage and early apoptosis in porcine oocytes. Collectively, we demonstrate that AAI exposure perturbs the oocyte meiotic progression and fertilization capacity via disruption of both nuclear maturation and cytoplasmic maturation of oocyte, which might be caused by the excessive oxidative stress-induced DNA damage and apoptosis.
    Keywords:  Aristolochic acid; DNA damage; fertilization capacity; oocyte maturation; oxidative stress
    DOI:  https://doi.org/10.18632/aging.101911
  27. Am J Physiol Renal Physiol. 2019 Apr 17.
    Cao W, Yuan Y, Liu X, Li Q, An X, Huang Z, Wu L, Zhang B, Zhang A, Xing C.
      Numerous studies have demonstrated that several mechanisms, including oxidative stress, DNA damage and inflammatory responses, are closely linked to cisplatin-induced nephrotoxicity. Adenosine, emerging as a key regulatory molecule, is mostly protective in the pathophysiology of inflammatory diseases. Previous study showed that some of the adenosine receptors led to renal protection against ischemia-reperfusion injury. However, these adenosine receptors agonists lack a useful therapeutic index due to cardiovascular side effects. We hypothesized that inhibition of adenosine kinase might exacerbate extracellular adenosine levels to reduce cisplatin-induced renal injury. In this study, pretreatment of adenosine kinase inhibitor ABT-702 could markedly attenuate cisplatin-induced acute kidney injury, tubular cell apoptosis, oxidative stress and inflammation in the kidneys. Consistent to in vivo results, the inhibition of ADK suppressed cisplatin-induced apoptosis, ROS production and inflammation in HK2 cells. Additionally, the protective effect of ADK inhibition was abolished by A1 or A2B adenosine receptor antagonist and enhanced by A2A or A3 adenosine receptor antagonist. Collectively, the results suggest that inhibition of ADK might increase the extracellular adenosine levels, which inhibited cisplatin-induced oxidative stress and inflammation via A1 and A2B adenosine receptors, finally suppressing cisplatin-induced cell apoptosis. Pharmacological therapies based on adenosine kinase will be of potential use for the therapy of cisplatin-induced nephrotoxicity.
    Keywords:  ABT-702; adenosine kinase; cisplatin nephrotoxicity
    DOI:  https://doi.org/10.1152/ajprenal.00385.2018
  28. Cell Death Dis. 2019 Apr 15. 10(5): 331
    Sui S, Zhang J, Xu S, Wang Q, Wang P, Pang D.
      (+)-JQ1 is an inhibitor of the tumor-driver bromodomain protein BRD4 and produces satisfactory effects because it efficiently increases apoptosis. Ferroptosis is an oxidative cell death program differing from apoptosis. Ferroptosis is characterized by high levels of iron and reactive oxygen species and has been confirmed to suppress tumor growth. In this study, BRD4 expression in cancer and its influence on the prognosis of cancer patients were analyzed using data from public databases. In addition, the effect of the BRD4 inhibitor (+)-JQ1 on ferroptosis was investigated via a series of in vitro assays. A nude mouse model was used to evaluate the function of (+)-JQ1 in ferroptosis in vivo. The potential mechanisms by which (+)-JQ1 regulates ferroptosis were explored. The results showed that BRD4 expression levels were higher in cancer tissues than in normal tissues and were related to poor prognosis in cancer patients. Furthermore, ferroptosis was induced under (+)-JQ1 treatment and BRD4 knockdown, indicating that (+)-JQ1 induces ferroptosis via BRD4 inhibition. Moreover, the anticancer effect of (+)-JQ1 was enhanced by ferroptosis inducers. Further studies confirmed that (+)-JQ1 induced ferroptosis via ferritinophagy, which featured autophagy enhancement by (+)-JQ1 and increased iron levels. Subsequently, the reactive oxygen species levels were increased by iron via the Fenton reaction, leading to ferroptosis. In addition, expression of the ferroptosis-associated genes GPX4, SLC7A11, and SLC3A2 was downregulated under (+)-JQ1 treatment and BRD4 knockdown, indicating that (+)-JQ1 may regulate ferroptosis by controlling the expression of ferroptosis-associated genes regulated by BRD4. Finally, (+)-JQ1 regulated ferritinophagy and the expression of ferroptosis-associated genes via epigenetic inhibition of BRD4 by suppressing the expression of the histone methyltransferase G9a or enhancing the expression of the histone deacetylase SIRT1. In summary, the BRD4 inhibitor (+)-JQ1 induces ferroptosis via ferritinophagy or the regulation of ferroptosis-associated genes through epigenetic repression of BRD4.
    DOI:  https://doi.org/10.1038/s41419-019-1564-7
  29. Biomed Pharmacother. 2019 Apr 16. pii: S0753-3322(18)37192-0. [Epub ahead of print]114 108855
    Ghosh S, Pal A, Ray M.
      The anti-carcinogenic effect of Methylglyoxal (MG) is well established. It generally targets malignant cells by affecting glycolysis and mitochondrial respiration with minimum or no toxicity to normal cells. In an initial study we have reported that MG can synergistically act with 5-Fluorouracil (5-FU) to decreases the number of MCF-7 breast cancer cells (Ghosh S, Pal A, Ray M, 2017). This finding prompted us to study the combination effect of MG and 5-FU extensively in both in vitro and in vivo. Induction of cell apoptosis and cell cycle arrest was systematically studied to reveal the mechanisms of synergy between 5-FU and MG. Our present study revealed that MG can synergistically act with 5-FU and can cause cell death via apoptosis and generated reactive oxygen species (ROS) in MCF-7 cells. Combination of 5-FU and MG resulted in more potent apoptosis induction as revealed by fluorescence microscopy using Hoechst 33342. In comparison to single drug treatment, the co-treatment also increased the number of cells in G0/G1 phase by downregulating the expression of CDK4 and CDK6 as compared to single drug treatment. Levels of Caspase 9 and poly (ADP-ribose) polymerase (PARP) were higher in combination treatment as compared to single drug treatment. These results clearly showed that 5-FU is more effective at lower doses in presence of MG in MCF-7 cells. In case of in vivo studies treatment of EAC (Ehrlich Ascites Carcinoma) bearing mice with MG in combination with 5-FU at various doses, demonstrated the same synergistic effect of MG with 5-FU. The combination study also exhibited tumor regression in BALB/c mouse 4T1 breast tumor model as well. We also clearly demonstrated that MG can decrease the cytotoxic side effects of 5-FU as indicated with acute and chronic toxicity studies and other biochemical analyses of blood and histological studies. Taken together, our results revealed that MG could be a potential candidate for combination therapy to reduce the toxicity burden of 5-FU without any toxic impact on host cells.
    Keywords:  5-Fluorouracil; Apoptosis; Breast cancer; Carcinoma; Methylglyoxal; Reactive oxygen species
    DOI:  https://doi.org/10.1016/j.biopha.2019.108855
  30. Exp Ther Med. 2019 May;17(5): 3734-3740
    Zhang L, Li C, Zhu Q, Li N, Zhou H.
      Cardiomyocyte apoptosis serves an important role in diabetic cardiomyopathy. Liraglutide, a glucagon-like peptide-1 analog, has been indicated to exert a cardioprotective effect. However, the role of liraglutide on cardiomyocyte apoptosis in hyperglycemia is not fully understood. The aim of the current study was to assess whether liraglutide protects against high glucose (HG)-induced cardiomyocyte apoptosis in vitro. Sprague-Dawley neonatal rat cardiomyocytes were cultured in Dulbecco's modified Eagle's medium, supplemented with 5.5 or 25 mmol/l D-glucose or 5.5 mmol/l D-glucose + 19.5 mmol/l mannitol, in the presence or absence of liraglutide (10 or 100 nmol/l). Cell viability was assessed via an MTT assay and early apoptosis rates were assessed via flow cytometry. Superoxide dismutase (SOD) activity and malondialdehyde (MDA) content in cell supernatants were measured. Bcl-2 associated X (Bax), B-cell lymphoma-2 (Bcl-2) and cleaved/full caspase-3 protein levels were determined via western blotting. The results revealed that liraglutide effectively inhibited the HG-induced increase in early apoptosis and MDA content and markedly increased SOD activity. Furthermore, liraglutide markedly inhibited the HG-induced increase in Bax and cleaved caspase-3 protein expression, and upregulated the expression of Bcl-2. The present study demonstrated that liraglutide suppressed HG-induced oxidative stress and cardiomyocyte apoptosis. Thus, the anti-apoptotic actions of liraglutide may be attributable, in part, to the inhibition of Bax, the inhibition of caspase-3 activation and the upregualtion of Bcl-2.
    Keywords:  apoptosis; cardiomyocyte; diabetic cardiomyopathy; glucagon-like peptide-1 analog; high glucose; liraglutide; oxidative stress
    DOI:  https://doi.org/10.3892/etm.2019.7388
  31. Oxid Med Cell Longev. 2019 ;2019 4957646
    He M, Shi W, Yu M, Li X, Xu J, Zhu J, Jin L, Xie W, Kong H.
      Acute lung injury (ALI) is a devastating critical disease characterized by diffuse inflammation and endothelial dysfunction. Increasing evidence, including from our laboratory, has revealed that the opening of ATP-sensitive potassium (KATP) channels has promising anti-inflammation and endothelial protection activities in various disorders. However, the impacts of KATP channels on ALI remain obscure. In this study, we used nicorandil (Nico), a classic KATP channel opener, to investigate whether opening of KATP channels could alleviate ALI with an emphasis on human pulmonary artery endothelial cell (HPAEC) modulation. The results showed that Nico inhibited lipopolysaccharide- (LPS-) induced inflammatory response, protein accumulation, myeloperoxidase activity, and endothelial injury. In vitro, Nico reduced LPS-induced HPAEC apoptosis and the expression of cleaved-caspase-3, caspase-9, and CCAAT/enhancer-binding protein homologous protein (CHOP). Additionally, Nico inhibited inflammation by suppressing monocyte-endothelial adhesion and decreasing the expression of proinflammatory proteins. Moreover, Nico restored the expression and the distribution of adherens junction vascular endothelial- (VE-) cadherin. Further, Nico abolished the increase in intracellular reactive oxygen species (ROS) and the activation of NF-κB and mitogen-activated protein kinase (MAPK) in HPAECs. Glibenclamide (Gli), a nonselective KATP channel blocker, abrogated the effects of Nico, implying that opening of KATP channels contributes to the relief of ALI. Together, our findings indicated that Nico alleviated LPS-induced ALI by protecting ECs function via preventing apoptosis, suppressing endothelial inflammation and reducing oxidative stress, which may be attributed to the inhibition of NF-κB and MAPK signaling pathways.
    DOI:  https://doi.org/10.1155/2019/4957646
  32. Biochem Biophys Res Commun. 2019 Apr 10. pii: S0006-291X(19)30633-3. [Epub ahead of print]
    Zhang D, Zhou Q, Huang D, He L, Zhang H, Hu B, Peng H, Ren D.
      Colorectal cancer (CRC) is one of the most common malignant neoplasms with high mortality worldwide. Oridonin, a diterpenoid isolated from the Chinese medicinal herb Rabdosia rubescens, has been proved to have anticancer effect on various types of cancer cells. However, the detailed mechanisms of oridonin in CRC cells remain unclear and if oridonin can overcome 5-FU resistance have not been investigated yet. In this study, we investigated the anticancer effect of oridonin in both 5-FU sensitive and resistant CRC cells and illuminated the underlying mechanisms. We showed that oridonin induced proliferation inhibition and caspase-dependent apoptosis in both 5-FU sensitive and resistant CRC cells. Oridonin induced reactive oxygen species (ROS) accumulation in both 5-FU sensitive and resistant CRC cells, which resulted in cell apoptosis as oridonin-induced apoptosis was almost abolished when cells were co-treated with the ROS scavenger N-acetyl-L-cysteine (NAC). Moreover, we found that oridonin induced CRC cell apoptosis via the c-Jun N-terminal kinase (JNK)/c-Jun pathway as oridonin activated JNK/c-Jun pathway and the JNK inhibitor SP600125 restored oridonin-induced apoptosis in CRC cells. Interestingly, when CRC cells were co-treated with NAC, the activation of JNK/c-Jun pathway induced by oridonin was nearly reversed, indicating that oridonin induced JNK/c-Jun pathway activation through the accumulation of ROS. Taken together, these data reveal that oridonin induces apoptosis through the ROS/JNK/c-Jun axis in both 5-FU sensitive and resistant CRC cells, suggesting that oridonin could be a potential agent for CRC treatment.
    Keywords:  Apoptosis; Caspase; Colorectal cancer; Oridonin; Reactive oxygen species; c-Jun N-Terminal kinase
    DOI:  https://doi.org/10.1016/j.bbrc.2019.04.011
  33. FASEB J. 2019 Apr 18. fj201801962RR
    Xu C, Xu J, Ji G, Liu Q, Shao W, Chen Y, Gu J, Weng Z, Zhang X, Wang Y, Gu A.
      X-ray repair cross-complementing group 1 (Xrcc1), a key DNA repair gene, plays a vital role in maintaining genomic stability and is highly expressed in the early stages of spermatogenesis, but the exact functions remain elusive. Here we generated primordial germ cell-specific Xrcc1 knockout (cXrcc1-/-) mice to elucidate the effects of Xrcc1 on spermatogenesis. We demonstrated that Xrcc1 deficiency results in infertility in male mice due to impaired spermatogenesis. We found that cXrcc1-/- mice exhibited smaller size of testes as well as lower sperm concentration and motility than the wild-type mice. Mechanistically, we demonstrated that Xrcc1 deficiency in primordial germ cells induced elevated levels of reactive oxygen species, mitochondria dysfunction, apoptosis, and loss of stemness of spermatogonial stem cells (SSCs) in testes. In Xrcc1-deficienct SSCs, elevated oxidative stress and mitochondrial dysfunction could be partially reversed by treatment with the antioxidant N-acetylcysteine (NAC), whereas NAC treatment did not restore the fertility or ameliorate the apoptosis caused by loss of Xrcc1. Overall, our findings provided new insights into understanding the crucial role of Xrcc1 during spermatogenesis.-Xu, C., Xu, J., Ji, G., Liu, Q., Shao, W., Chen, Y., Gu, J., Weng, Z., Zhang, X., Wang, Y., Gu, A. Deficiency of X-ray repair cross-complementing group 1 in primordial germ cells contributes to male infertility.
    Keywords:   spermatogenesis; mitochondrial dysfunction; oxidative stress
    DOI:  https://doi.org/10.1096/fj.201801962RR
  34. Toxicol Lett. 2019 Apr 10. pii: S0378-4274(19)30097-9. [Epub ahead of print]
    Abdel-Zaher AO, Abd-Ellatief RB, Aboulhagag NA, Farghaly HSM, Al-Wasei FMM.
      This study explored the role of gasotransmitters in lead-induced nephrotoxicity. Long-term exposure of rats to lead resulted in its accumulation in kidney. The accumulated metal impaired kidney function and structure. Lead intoxication resulted in oxidative stress, inflammation and apoptosis in kidney. In addition, it resulted in nitric oxide (NO) overproduction and decrease in hydrogen sulfide (H2S) level and heme oxygenase (HO-1) concentration in kidney. Inhibition of NO overproduction by L-N(G)-nitroarginine methyl ester (L-NAME) and increasing of H2S level by sodium hydrosulfide (Na HS) and CO level by carbon monoxide-releasing molecule-A1 (CORM-A1) inhibited lead-induced impairment of kidney function and structure. These agents inhibited lead-intoxication induced oxidative stress, inflammation, apoptosis, nitrosative stress and reduction of H2S level and HO-1 concentration. Also, concomitant treatment with these agents inhibited lead intoxication-induced increase in protein expressions of inducible NO synthase (iNOS), tumor necrosis factor-alpha (TNF-α), interleukin-1beta (IL-1β) and caspase-3 as well as decrease in protein expressions of HO-1 and cystathionine- γ-lyase (CSE) in kidney. The NO donor, L-arginine and the H2S and CO biosynthesis inhibitors, trifluoro-DL-alanine and zinc deutroporphyrin, respectively produced opposite effects and aggravated the toxic effects of lead. These results demonstrate, for the first time, that gasotransmitters play an important role in lead-induced nephrotoxicity.
    Keywords:  Apoptosis; Gasotransmitters; Inflammation; Lead; Nephrotoxicity; Oxidative stress
    DOI:  https://doi.org/10.1016/j.toxlet.2019.04.012
  35. Nutrients. 2019 Apr 16. pii: E857. [Epub ahead of print]11(4):
    Hwang BB, Chang MH, Lee JH, Heo W, Kim JK, Pan JH, Kim YJ, Kim JH.
      Accumulation of reactive oxygen species (ROS) in response to excess alcohol exposure is a major cause of gut barrier disruption and lipopolysaccharide (LPS)-induced hepatic inflammation, as well as liver steatosis and apoptosis. This study was designed to investigate protective effects of the cricket Gryllus bimaculatus, an edible insect recognized by the Korea Food and Drug Administration, against acute alcoholic liver damage in mice. Administration of G. bimaculatus extracts (GBE) attenuated alcohol-induced steatosis and apoptotic responses in the liver and intestinal permeability to bacterial endotoxin. These protective effects were associated with suppression of ROS-mediated oxidative stress in both the liver and small intestine. Furthermore, in vivo and in vitro studies revealed that GBE inhibits LPS-induced Kupffer cell activation and subsequent inflammatory signaling. Importantly, the protective effects of GBE were more potent than those of silymarin, a known therapeutic agent for alcoholic liver diseases.
    Keywords:  Gryllus bimaculatus; alcoholic liver diseases; intestinal permeability; reactive oxygen species
    DOI:  https://doi.org/10.3390/nu11040857
  36. Hum Exp Toxicol. 2019 Apr 16. 960327119842613
    Zaki SM, Algaleel WA, Imam RA, Abdelmoaty MM.
      The cardiotoxic adverse effect of doxorubicin (DOX) is the major factor limiting its use. Recently, mesenchymal stem cells (MSCs) have been implicated in the preclinical studies of treatment of DOX-induced cardiotoxicity. The question is MSCs pretreated with platelet-rich plasma (PRP) have a better influence on DOX-induced cardiotoxicity compared to the influence of MSCs alone. Twenty-four Wistar rats were categorized into control, DOX-treated, MSC-treated, and PRP/MSC-treated groups. DOX was injected for two consecutive weeks. Light microscopic, biochemical markers (interleukin 10 (IL-10), tumor necrosis factor alpha (TNF-α), and creatine kinase-MB (CK-MB)), immunohistochemical (Bax, Bcl2, vascular endothelial growth factor (VEGF), and cardiac troponin-I (CT-I)), and oxidative/antioxidative markers (malondialdehyde (MDA)/superoxide dismutase (SOD)) were measured. Degenerative cardiac changes were detected in the DOX-treated group with complete loss of the architecture and coagulative necrosis. These changes were accompanied with the elevation of the serum level of CK-MB and loss of CT-I immunoreactivity. The major factors in the DOX-induced cardiotoxicity were the oxidative stress (elevated MDA/decreased SOD), inflammation (elevated TNF-α/decreased IL-10), and cardiac apoptosis (lower Bcl2, higher Bax, and lower Bcl2/Bax ratio). MSCs and PRP/MSCs attenuate DOX-induced cardiotoxicity. Better attenuation was observed in the PRP/MSC-treated group. PRP/MSC combination reduced greatly the MDA and TNF-α and increased IL-10, Bcl2/Bax ratio, and VEGF. PRP had no significant influence over the Bcl2, Bax, and SOD. In conclusion, DOX in its toxic dose induced myocardial injury. This destructive effect is related to oxidative stress, inflammation, and cardiac apoptosis. PRP/MSC possesses a better attenuation over the DOX-induced toxicity compared to MSC alone.
    Keywords:  Platelet-rich plasma; cardiotoxicity; doxorubicin; mesenchymal stem cell
    DOI:  https://doi.org/10.1177/0960327119842613
  37. Neuropharmacology. 2019 Apr 12. pii: S0028-3908(19)30120-0. [Epub ahead of print]
    Zhou L, Cheng Y.
      Oxidative stress and autophagy are involved in the pathogenesis of Parkinson's disease.The relationship between oxidative stress and autophagy is a hot spot of scientific research. Alpha-lipoic acid (ALA) is a natural antioxidant. ALA has been reported to reduce oxidative stress and apoptosis in PD models, but its role in autophagy regulation of PD has been reported very little. In this study, we investigated the protective effects of ALA on 6-OHDA induced neurotoxicity, and explored the potential mechanisms associated with the crosstalk between oxidative stress and autophagy. Our results showed that 6-OHDA induced accumulation of ROS in SH-SY5Y cells, accompanied by excessive autophagy and apoptosis. ALA protected against 6-OHDA induced neuronal death through inhibition of oxidative stress and autophagy. Furthermore, we found that ALA inhibited AMPK phosphorylation while activated mTOR phosphorylation thereby blocking AMPK/mTOR signaling pathway involved autophagy. In conclusion, ALA alleviated 6-OHDA induced cell injury possibly by inhibiting autophagy mediated by AMPK/mTOR pathway.
    Keywords:  Parkinson’s disease (PD);; autophagy;; oxidative stress; α-lipoic acid (ALA);
    DOI:  https://doi.org/10.1016/j.neuropharm.2019.04.009
  38. Endocr Relat Cancer. 2019 Jun 01. pii: ERC-18-0517. [Epub ahead of print]26(6): 551-563
    Saini S, Sripada L, Tulla K, Qiao G, Kunda N, Maker AV, Prabhakar BS.
      ATC is an aggressive disease with limited therapeutic options due to drug resistance. TRAIL is an attractive anti-cancer therapy that can trigger apoptosis in a cancer cell-selective manner. However, TRAIL resistance is a major clinical obstacle for its use as a therapeutic drug. Previously, we demonstrated that MADD is a cancer cell pro-survival factor that can modulate TRAIL resistance. However, its role, if any, in overcoming TRAIL resistance in ATC is unknown. First, we characterized ATC cell lines as either TRAIL resistant, TRAIL sensitive or moderately TRAIL sensitive and evaluated MADD expression/cellular localization. We determined the effect of MADD siRNA on cellular growth and investigated its effect on TRAIL treatment. We assessed the effect of combination treatment (MADD siRNA and TRAIL) on mitochondrial membrane potential (MMP) and reactive oxygen species (ROS) levels. The effect of combination treatment on tumor growth was assessed in vivo. We found increased levels of MADD in ATC cells relative to Nthy-ori 3-1. MADD protein localizes in the cytosol (endoplasmic reticulum and Golgi body) and membrane. MADD knockdown resulted in spontaneous cell death that was synergistically enhanced when combined with TRAIL treatment in otherwise resistant ATC cells. Combination treatment resulted in a significant reduction in MMP and enhanced generation of ROS indicating the putative mechanism of action. In an orthotopic mouse model of TRAIL-resistant ATC, treatment with MADD siRNA alone reduced tumor growth that, when combined with TRAIL, resulted in significant tumor regressions. We demonstrated the potential clinical utility of MADD knockdown in sensitizing cells to TRAIL-induced apoptosis in ATC.
    Keywords:  TRAIL; siRNA; thyroid cancer; tumor
    DOI:  https://doi.org/10.1530/ERC-18-0517
  39. Int J Mol Sci. 2019 Apr 03. pii: E1654. [Epub ahead of print]20(7):
    Essid SM, Bevington A, Brunskill NJ.
      The repair capacity of progenitor skeletal muscle satellite cells (SC) in Type 1 diabetes mellitus (T1DM) is decreased. This is associated with the loss of skeletal muscle function. In T1DM, the deficiency of C-peptide along with insulin is associated with an impairment of skeletal muscle functions such as growth, and repair, and is thought to be an important contributor to increased morbidity and mortality. Recently, cholesterol-lowering drugs (statins) have also been reported to increase the risk of skeletal muscle dysfunction. We hypothesised that C-peptide activates key signaling pathways in myoblasts, thus promoting cell survival and protecting against simvastatin-induced myotoxicity. This was tested by investigating the effects of C-peptide on the L6 rat myoblast cell line under serum-starved conditions. Results: C-peptide at concentrations as low as 0.03 nM exerted stimulatory effects on intracellular signaling pathways-MAP kinase (ERK1/2) and Akt. When apoptosis was induced by simvastatin, 3 nM C-peptide potently suppressed the apoptotic effect through a pertussis toxin-sensitive pathway. Simvastatin strongly impaired Akt signaling and stimulated the reactive oxygen species (ROS) production; suggesting that Akt signaling and oxidative stress are important factors in statin-induced apoptosis in L6 myoblasts. The findings indicate that C-peptide exerts an important protective effect against death signaling in myoblasts. Therefore, in T1DM, the deficiency of C-peptide may contribute to myopathy by rendering myoblast-like progenitor cells (involved in muscle regeneration) more susceptible to the toxic effects of insults such as simvastatin.
    Keywords:  Type 1 diabetes; myotoxicity; proinsulin C-peptide; simvastatin
    DOI:  https://doi.org/10.3390/ijms20071654
  40. Alcohol. 2019 Apr 11. pii: S0741-8329(18)30251-9. [Epub ahead of print]
    Vaghef L, Farajdokht F, Erfani M, Majdi A, Sadigh-Eteghad S, Karimi P, Shotorbani SS, Vafaee MS, Mahmoudi J.
      The present study investigates the potential neuroprotective effect of cerebrolysin (CBL), a combination of neurotrophic factors, on the cognitive and biochemical alterations induced by chronic ethanol administration in rats. The animals were divided into 5 groups as follows: Control, ethanol (4 g/kg, for 30 days) plus normal saline (Ethanol + NS), ethanol plus CBL 1 ml/kg (Ethanol + CBL 1), ethanol plus CBL 2.5 ml/kg (Ethanol + CBL 2.5), and ethanol plus CBL 5 ml/kg (Ethanol + CBL 5). Morris water maze (MWM) test was performed to assess cognitive impairment. The status of the lipid peroxidation marker (MDA), antioxidant capacity as well as alterations of the apoptotic factors such as Bcl-2, BAX, and cleaved-caspase 9 and 3 were evaluated in the hippocampus. The results showed that CBL treatment not only normalized the increased MDA levels in the alcoholic rats and enhanced antioxidant defense but also reduced Bax/Bcl-2 ratio and cleaved-caspase 9 and 3 in the hippocampus. These results were parallel with improvement in the spatial memory performance in the MWM test. The findings of the present study provide evidence for the promising therapeutic effect of CBL in chronic ethanol consumption through counteracting oxidative stress and apoptosis markers.
    Keywords:  Apoptosis; Cerebrolysin; Ethanol; Oxidative stress; Spatial memory
    DOI:  https://doi.org/10.1016/j.alcohol.2019.03.005
  41. J Cell Mol Med. 2019 Apr 14.
    Zhou F, Mei J, Yuan K, Han X, Qiao H, Tang T.
      Increasing evidence indicates that osteoarthritis (OA) is a musculoskeletal disease affecting the whole joint, including both cartilage and subchondral bone. Reactive oxygen species (ROS) have been demonstrated to be one of the important destructive factors during early-stage OA development. The objective of this study was to investigate isorhamnetin (Iso) treatment on osteoclast formation and chondrocyte protection to attenuate OA by modulating ROS. Receptor activator of nuclear factor-kappa B ligand (RANKL) was used to establish the osteoclast differentiation model in bone marrow macrophages (BMMs) in vivo. H2 O2 was used to induce ROS, which could further cause chondrocyte apoptosis. We demonstrated that Iso suppressed RANKL-induced ROS generation, which could mediate osteoclastogenesis. Moreover, we found that Iso inhibited osteoclast formation and function by suppressing the expression of osteoclastogenesis-related genes and proteins. We proved that Iso inhibited RANKL-induced activation of mitogen-activated protein kinase activation of mitogen-activated protein kinase (MAPK), nuclear factor-kappa B (NF-κB) and AKT signalling pathways in BMMs. In addition, Iso inhibited ROS-induced chondrocyte apoptosis by regulating apoptosis-related proteins. Moreover, Iso was administered to an anterior cruciate ligament transection (ACLT)-induced OA mouse model. The results indicated that Iso exerted beneficial effects on inhibiting excessive osteoclast activity and chondrocyte apoptosis, which further remedied cartilage damage. Overall, our data showed that Iso is an effective candidate for treating OA.
    Keywords:  ROS; apoptosis; isorhamnetin; osteoarthritis; osteoclast
    DOI:  https://doi.org/10.1111/jcmm.14333
  42. Med Sci Monit. 2019 Apr 15. 25 2764-2776
    Wang P, Sun J, Lv S, Xie T, Wang X.
      BACKGROUND We investigated whether apigenin could mitigate myocardial reperfusion injury in rats, and a possible mechanism was proposed. MATERIAL AND METHODS The I-R injury model was established in rats along with a sham group as control, and the expressions of microRNA-15b (miR-15b), JAK2, and p-JAK2 in the myocardia of the 2 groups were detected. Apoptosis and reactive oxygen species (ROS) were also detected. Rats in the I-R injury model were divided into 3 groups in vivo: the 1I-R group, the 2I-R+solvent group, and the 3I-R+apigenin group. Expression of miR-15b, JAK2, p-JAK2, STAT3, and p-STAT3 in the myocardia of the 3 groups were detected. ROS content, apoptosis, MDA content, SOD, and CAT activities were detected. Rat myocardial H9C2 cells were cultured in vitro and divided into 5 treatment groups in vitro; expressions of miR-15b, JAK2, p-JAK2, STAT3, and p-STAT3 in H9C2 cells were detected, and the apoptosis and ROS content were detected by flow cytometry. RESULTS We found that the increased miR-15b expression during myocardial I-R injury in rats downregulated the expression of JAK2 and activity of the JAK2-STAT3 pathway, promoted myocardial apoptosis and ROS production, and aggravated myocardial I-R injury. Apigenin treatment can downregulate miR-15b expression, increase the expression of JAK2 and the activity of JAK2-STAT3 pathway, reduce myocardial apoptosis and ROS production, and alleviate myocardial I-R injury. CONCLUSIONS Api treatment downregulated the expression of miR-15b and upregulated the expression of JAK2 and the activity of the JAK2-STAT3 pathway, thereby alleviating myocardial I-R injury, cardiomyocyte apoptosis, and ROS production in vitro.
    DOI:  https://doi.org/10.12659/MSM.912014
  43. Food Res Int. 2019 Jun;pii: S0963-9969(19)30116-4. [Epub ahead of print]120 52-61
    Gómez LJ, Gómez NA, Zapata JE, López-García G, Cilla A, Alegría A.
      The antioxidant capacity of red tilapia viscera hydrolysates (RTVH) with different degrees of hydrolysis (DH) as well as their ultrafiltration membrane fractions, were analyzed using different chemical assays. Their protective effects against oxidative stress were evaluated using H2O2-stressed human intestinal differentiated Caco-2. The highest antioxidant capacity was obtained with a DH of 42.5% (RTVH-A) and its <1 kDa fraction (FRTVH-V). RTVH-A and FRTVH-V did not show cytotoxic effects at a concentration of ≤0.5 mg/mL,prevented the decrease in cell viability, and suppressed intracellular reactive oxygen species (ROS) accumulation induced by H2O2. However, pretreatment with RTVH-A after adding H2O2, showed a greater decrease in glutathione levels. Moreover, FRTVH-V allowed for a recovery close to that of control levels of cell proportions in the G1 and G2/M cell cycle phases; and a decrease in the cell proportion in late apoptosis. These results suggest that RTVH-A and FRTVH-V can be beneficial ingredients with antioxidant properties and can have protective effects against ROS-mediated intestinal injuries.
    Keywords:  Alcalase; Antioxidant activity; Biocatalysis; Cytoprotective effect; Oxidative stress; Protein hydrolysates
    DOI:  https://doi.org/10.1016/j.foodres.2019.02.029
  44. Cancer Cell Int. 2019 ;19 85
    Karna KK, Choi BR, You JH, Shin YS, Soni KK, Cui WS, Lee SW, Kim CY, Kim HK, Park JK.
      Background: DA-9401 was prepared as a mixture of Chinese medicinal herb extracts from roots of Morinda officinalis How (Rubiaceae), outer scales of Allium cepa L. (Liliceae) and seeds of Cuscuta chinensis Lamark (Convolvulaceae). The present study was designed to investigate the possible protective role of DA-9401 in adriamycin (ADR)-induced testicular toxicity associated with oxidative stress, endoplasmic reticulum (ER) stress, and apoptosis.Methods: Fifty healthy 8-week-old male Sprague-Dawley rats were equally divided into five groups. The first CTR group was treated with normal saline 2 ml/day by gavage. The second was treated with DA-100 (DA-9401 100 mg/kg/day). The third (ADR) group received ADR (2 mg/kg/once a week) intraperitoneally, while the combination of ADR and DA-9401 was given to the fourth ADR + DA-100 (100 mg/kg/day p.o) group and fifth ADR + DA-200 (200 mg/kg/day p.o) group. At the end of the 8-week treatment period, body weight, reproductive organ weights, fertility rate, pups per female were recorded, and serum were assayed for hormone concentrations. Tissues were subjected to semen analysis, histopathological changes, interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), oxidative stress markers and expression levels of endoplasmic reticulum (ER) stress markers, apoptosis markers, tight junction protein markers, steroidogenic acute regulatory protein (StAR), cation channel of sperm (CatSper) and glycogen synthase kinase-3 (GSK-3) by western blot.
    Results: DA-9401 administration to ADR-treated rats significantly decreased serum luteinizing hormone (LH) and follicle-stimulating hormone (FSH) levels, interleukin-6, TNF-α, MDA level, ROS/RNS level, ER stress response protein levels, tunnel positive cells, cleaved caspase-3, and Bax/Bcl2 ratio. Moreover, pretreatment with DA-9401 significantly increased body weight, reproductive organ weights, fertility rate, pups per female, Johnsen's score, spermatogenic cell density, sperm count and sperm motility, serum testosterone concentration, testicular superoxide dismutase (SOD), catalase, glutathione peroxidase (GPx), tight junction protein markers, star protein level, CatSper, and GSK-3 level.
    Conclusions: ADR treatment can markedly impair testicular function and induce testicular cell death presumably by causing significant changes in oxidative stress, ER stress, and mitochondrial pathway. DA-9401 exerts beneficial effects against oxidative stress, ER stress, and mitochondria-mediated cell death pathway in testis tissue by up-regulating expression levels of tight junction protein markers, steroidogenic acute regulatory protein, GSK-3 alpha, and cation channels of sperm.
    Keywords:  Adriamycin (ADR); Apoptosis; Blood-testis barrier (BTB); Cation channel of sperm (CatSper); DA-9401; Endoplasmic reticulum (ER) stress; Glycogen synthase kinase-3 (GSK-3); Oxidative stress; Steroidogenic acute regulatory protein (StAR)
    DOI:  https://doi.org/10.1186/s12935-019-0805-2
  45. Colloids Surf B Biointerfaces. 2019 Mar 29. pii: S0927-7765(19)30215-2. [Epub ahead of print]179 421-428
    Young Park S, Jin Kim Y, Park G, Kim HH.
      Combination therapy remains a promising approach to ameliorate cerebral ischemia injury. Nevertheless, the primary mechanism of the neuroprotective properties of Dictyopteris divaricata extract-capped gold nanoparticles (DD-GNPs) is not completely understood. DD-GNPs displayed maximum absorption at 525 nm and a diameter of 62.6 ± 1.2 nm, with a zeta potential value of -26.1 ± 0.6 mV. High resolution-transmission electron microscopy confirmed the spherical shape and average diameter (28.01 ± 2.03 nm). Crystalline structure and gold nanoparticle synthesis of DD-GNPs were determined by X-ray powder diffraction, and the presence of elemental gold was confirmed by energy-dispersive X-ray spectroscopy and Fourier transform-infrared spectroscopy. We examined the neuroprotective properties of DD-GNPs and explored their potential mechanisms in human SH-SY5Y neuroblastoma cells treated with oxygen and glucose deprivation/reoxygenation (OGD/R). We found that DD-GNPs inhibited OGD/R-induced release of lactate dehydrogenase (LDH), loss of cell viability, and production of reactive oxygen species. This neuroprotection was accompanied by regulation of apoptosis-related proteins, as indicated by decreased levels of cleaved-caspase-3, cleaved-PARP, cleaved-caspase-9, p53, p21, and Bax, as well as an increased level of Bcl-2. Notably, the neuroprotective effects of DD-GNPs were partially abolished by HO-1, NQO1, Nrf2, and AMPK knockdown. Our results established that DD-GNPs effectively attenuated OGD/R-stimulated neuronal injury, as evidenced by reduced neuronal injury. Even though the accumulating evidence has indicated the low toxicity and minimal side effects of GNPs, experimental clinical trials of DD-GNPs are still limited because of the lack of knowledge regarding the effects of DD-GNPs as neuroprotective agents against neurodegenerative diseases.
    Keywords:  AMPK; DD-GNPs; Dictyopteris divaricate; Neuroprotection; Nrf2; OGD/R
    DOI:  https://doi.org/10.1016/j.colsurfb.2019.03.066
  46. J Shoulder Elbow Surg. 2019 Apr 10. pii: S1058-2746(19)30073-4. [Epub ahead of print]
    Mifune Y, Inui A, Muto T, Nishimoto H, Kataoka T, Kurosawa T, Yamaura K, Mukohara S, Niikura T, Kokubu T, Kuroda R.
      BACKGROUND: Most rotator cuff tears are the result of age-related degenerative changes, but the mechanisms underlying these changes have not been reported. Recently, advanced glycation end products (AGEs) have been regarded as an important factor in senescence. Therefore, we hypothesized that AGEs would have detrimental effects on rotator cuff-derived cells. In this study, we investigated the influence of AGEs on rotator cuff-derived cells in vitro and ex vivo.METHODS: Rotator cuff-derived cells were obtained from human supraspinatus tendons. The cells were cultured in the following media: (1) regular medium with 500 μg/mL AGEs (High-AGEs), (2) regular medium with 100 μg/mL AGEs (Low-AGEs), and (3) regular medium alone (Control). Cell viability, secretion of vascular endothelial growth factor, and the expressions of hypoxia-inducible factor-1α, reactive oxygen species, and apoptosis were assessed after cultivation. An ex vivo tissue culture with AGEs was also performed to measure the tensile strength.
    RESULTS: Cell viability in the High-AGEs group was significantly suppressed relative to that in the Controls. The amount of vascular endothelial growth factor secretion was significantly greater in the High- and Low-AGEs groups than in the Controls. Immunofluorescence stain demonstrated enhancement of hypoxia-inducible factor-1α and reactive oxygen species expressions and cell apoptosis in the High- and Low-AGEs groups relative to that in the Controls. In ex vivo mechanical testing, tensile strength was significantly higher in the Control group than in the AGEs groups.
    DISCUSSION: These results indicated that AGEs caused age-related degenerative rotator cuff changes. The reduction of AGEs might prevent rotator cuff senescence-related degeneration.
    Keywords:  Advanced glycation end products; apoptosis; degeneration; hypoxia-inducible factor-1α; reactive oxygen species; rotator cuff; vascular endothelial growth factor
    DOI:  https://doi.org/10.1016/j.jse.2019.01.022
  47. Exp Eye Res. 2019 Apr 16. pii: S0014-4835(18)30239-2. [Epub ahead of print]
    Ali SA, Zaitone SA, Dessouki A, Ali AA.
      Pregabalin is the first drug to receive FDA approval for treating diabetic neuropathic pain. This study investigated the neuroprotective effect of pregabalin in an experimental model of diabetic retinopathy and tested some possible mechanisms underlying the putative neuroprotective effect. Male Wistar rats received streptozotocin (45 mg/kg) to induce type 1 diabetes mellitus. After two weeks, a course of pregabalin (3, 10 and 30 mg/kg) has been launched for five consecutive weeks. Retinal expression of interleukin-1β (IL-1β) and tumor necrosis factor-α (TNF-α) was estimated by real-time PCR and retinal glutamate content was also estimated. Further, retinal caspase-3 immunoblotting and DNA fragmentation assays determined the degree of apoptosis. Pregabalin improved histopathological abnormalities in diabetic retinas and suppressed the diabetes-enhanced retinal expression of IL-1β, TNF-α, CD11b (a surface marker for microglia) while attenuated expression of caspase-3 and DNA fragmentation versus the diabetic group. In addition, diabetic rats treated with pregabalin displayed reductions in retinal glutamate, nitric oxide and malondialdehyde (MDA) and enhanced reduced glutathione (GSH) content versus the diabetic controls. Furthermore, pregabalin enhanced the histopathological picture and reduced fibrosis in the optic nerve of diabetic rats in addition to suppression of the content of the glia fibrillary acidic protein. The findings provide the first evidence demonstrating that pregabalin alleviates retinal neuroinflammation, apoptosis and oxidative stress in an experimental type 1 diabetes mellitus. Therefore, pregabalin might serve as a potential therapy for retinopathy after adequate clinical research.
    Keywords:  Apoptosis; Diabetic retinopathy; Glutamate; Pregabalin; Rat; Retinal glia
    DOI:  https://doi.org/10.1016/j.exer.2019.04.014
  48. Biotech Histochem. 2019 Apr 17. 1-7
    Fırat F, Özgül M, Türköz Uluer E, Inan S.
      Cancer is a common cause of death worldwide. Approximately 80% of cancer patients use complementary or alternative medicines for treatment. Caffeic acid phenethyl ester (CAPE), the main active component of propolis, exhibits cytotoxic, antiproliferative and anti-cancer effects. Despite its anticancer effects CAPE exhibits no known harmful effects toward normal cells. We investigated the effects of CAPE on angiogenesis, apoptosis and oxidative stress using MDA MB-231, N2a and COLO 320 cell lines and CAPE treatments at 24 and 48 h. A two dimensional cell culture system was used and the findings were evaluated by an indirect immunohistochemical method and H-scores were calculated. CAPE was effective for all three cancer cell lines. After 24 and 48 h, we found a significant decrease in live cells and increased stress in the cells based on e-NOS and i-NOS levels.
    Keywords:  Angiogenesis; CAPE; apoptosis; caffeic acid phenethyl ester; cancer; oxidative stress
    DOI:  https://doi.org/10.1080/10520295.2019.1589574
  49. Chem Sci. 2019 Mar 07. 10(9): 2778-2784
    Wei Z, Liang P, Xie J, Song C, Tang C, Wang Y, Yin X, Cai Y, Han W, Dong X.
      Herein, a nano-integrated strategy was used to combine an anti-angiogenic agent sorafenib and a photosensitizer chlorin e6 to form carrier-free multifunctional nanoparticles (SC NPs) for synergetic anti-angiogenic therapy and phototherapy. SC NPs (diameter, ∼152 nm) presented excellent water dispersity and passive targeting ability towards tumor sites in vivo based on the enhanced permeability and retention (EPR) effect, which could be monitored by fluorescence imaging. Besides, SC NPs exhibited effective reactive oxygen species (ROS) generation and photothermal conversion abilities for both photodynamic therapy (PDT) and photothermal therapy (PTT). At a rather low dosage (200 μg kg-1) and illumination with laser (660 nm, 500 mW cm-2), SC NPs could attack tumor tissues by killing the internal tumor cells via mild phototherapy, simultaneously cutting off the external nutrient and oxygen supplements of the tumor cells via anti-angiogenesis. Besides, oxygen consumption in the PDT process may be combined with anti-angiogenic therapy to further cause cell apoptosis by tumor starvation. In addition to the highly efficient therapeutic effect in vivo, SC NPs possessed excellent biosafety and biocompatibility, making them promising for fluorescence imaging-guided synergetic anti-angiogenic therapy and phototherapy in clinic.
    DOI:  https://doi.org/10.1039/c8sc04123g
  50. World Neurosurg. 2019 Apr 10. pii: S1878-8750(19)31042-3. [Epub ahead of print]
    Akkaya E, Evran Ş, Çalış F, Çevik S, Hanımoğlu H, Seyithanoğlu MH, Katar S, Karataş E, Koçyiğit A, Sağlam MY, Hatiboğlu MA, Kaynar MY.
      OBJECTIVE: Verapamil-a calcium-channel blocker-has shown promising results on cerebral vasospasm. However, it has not yet been accepted for the treatment or prevention purposes because of the associated side-effects. Although effective results of nimodipine and nicardipine's intrathecal administration are well-known, intrathecal verapamil has not been considered earlier. We used an experimental subarachnoid hemorrhage induced vasospasm model for the evaluation of vasodilator and neuroprotective effects of intrathecal verapamil.METHODS: 24 Sprague-Dawley rats were randomly divided into 3 groups. Group 1(sham), group 2(subarachnoid hemorrhage), and group 3(verapamil). Double hemorrhage method was used. Group 2 did not receive any treatment. Verapamil (rhEPO;Eporon, Dem Ilac, Turkey) at a dose of 1,000 μg/kg was given intrathecally to group 3 rats. The animals were sacrified on day 7 of the procedure. Arterial wall thicknesses and lumen diameters in the basilar arterial cross-sectional areas, endothelin-1 serum levels, oxidative stress index, and apoptosis were measured in all groups.
    RESULTS: In the verapamil group, the wall thickness, endothelin-1 levels, oxidative stress index, and apoptosis were found significantly lower than in the subarachnoid hemorrhage group, but the lumen diameter was found to be greater. Intrathecal verapamil was found to decrease vasospasm parameters and apoptosis and increase the antioxidant and antiapoptotic pathways.
    CONCLUSIONS: Our findings suggest that intrathecal verapamil can prevent vasospasm, oxidative stress, and apoptosis following experimental subarachnoid hemorrhage.
    Keywords:  Aneurysm; apoptosis; endothelin-1; intrathecal verapamil; oxidative stress; subarachnoid hemorrhage; vasospasm
    DOI:  https://doi.org/10.1016/j.wneu.2019.04.050
  51. Exp Ther Med. 2019 May;17(5): 3848-3858
    Guo C, Wang L, Zhao Y, Jiang B, Luo J, Shi D.
      The novel bromophenol derivative, 3-(3-bromo-5-methoxy-4-(3-(piperidin-1-yl)propoxy)benzylidene)-N-(4-bromophenyl)-2-oxoindoline-5-sulfonamide (BOS-93), was synthesized in the CAS Key Laboratory of Experimental Marine Biology, Institute of Oceanology, Chinese Academy of Sciences (Qingdao, China). Experimental studies have demonstrated that it could induce apoptosis and autophagy in human A549 lung cancer cells, and it could also inhibit tumor growth in human A549 lung cancer xenograft models. In the present study, the molecular pathways underlying these effects were identified. The results demonstrated that BOS-93 could inhibit cell proliferation in A549 cells and block A549 cells at the G0/G1 phase. Furthermore, BOS-93 could induce apoptosis, activate caspase-3 and poly ADP ribose polymerase, and increase the B cell lymphoma (Bcl)-2 associated X protein/Bcl-2 ratio. Notably, BOS-93 could also induce autophagy in A549 cells. BOS-93-induced autophagy was confirmed by detecting light chain 3 (LC3)-I/LC3-II conversion and increasing expression of beclin1 and autophagy-related gene 14. Notably, BOS-93-induced autophagy could be inhibited by the autophagy inhibitor 3-MA. Flow cytometry, transmission electron microscopy (TEM) and western blot analysis indicated that BOS-93 induced apoptosis and autophagy activities by deactivating phosphoinositide 3-kinase/protein kinase B/mechanistic target of rapamycin and activating the mitogen-activated protein kinase signaling pathway. The present findings indicated that BOS-93 might be a novel anti-cancer agent for treatment of human lung cancer.
    Keywords:  apoptosis; autophagy; bromophenol; phosphoinositide 3-kinase/protein kinase B/mechanistic target of rapamycin signaling; reactive oxygen species
    DOI:  https://doi.org/10.3892/etm.2019.7402
  52. Biomed Pharmacother. 2019 Apr 10. pii: S0753-3322(19)30759-0. [Epub ahead of print]114 108825
    Zhang L, Li S, Wang R, Chen C, Ma W, Cai H.
      Large tumor suppressor 2 (LATS2), an important mediator of the cell apoptotic response pathway, has been linked to the progression of several cancers. Here, we described the molecular feature of LATS2 as a novel antitumor factor in liver cancer cells in vitro. Western blotting was used to detect the expression of LATS2 and its downstream factors. ELISA, immunofluorescence, and flow cytometry were used to evaluate the alterations of mitochondrial function in response to LATS2 overexpression. Adenovirus-loaded LATS2 and siRNA against DRP1 were transfected into liver cancer cells to overexpress LATS2 and knockdown DRP1 expression, respectively. The results of the present study demonstrated that overexpression of LATS2 was closely associated with more liver cancer cell death. Mechanistically, LATS2 overexpression increased the expression of DRP1, and DRP1 elevated mitochondrial division, an effect that was accompanied by mitochondrial dysfunction, including mitochondrial membrane potential reduction, mitochondrial respiratory complex downregulation, mitochondrial cyt-c release into the nucleus and mitochondrial oxidative injury. Moreover, LATS2 overexpression also initiated mitochondrial apoptosis, and this process was highly dependent on DRP1-related mitochondrial division. Molecular investigations demonstrated that LATS2 modulated DRP1 expression via the Wnt/β-catenin pathway. Inhibition of the Wnt/β-catenin pathway pregented LATS2-mediated DRP1 upregulation, ultimately sustaining mitochondrial function and cell viability in the presence of LATS2 overexpression. Altogether, the above data identify LATS2-Wnt/β-catenin/DRP1/mitochondrial division as a novel anticancer signaling pathway promoting cancer cell death, which might be an attractive therapeutic target for the treatment of hepatocellular carcinoma.
    Keywords:  DRP1; LATS2; Mitochondria; Wnt/β-catenin pathway
    DOI:  https://doi.org/10.1016/j.biopha.2019.108825
  53. Nat Prod Res. 2019 Apr 16. 1-3
    Castillo-Bautista CM, Torres-Tapia LW, Lagunas-Martínez A, Contreras-Ochoa CO, Peraza-Sanchez SR, Moo-Puc R.
      Secundiflorol G (SG) is an isoflavan isolated from the root bark of Aeschynomene fascicularis, a Mayan medicinal plant used to treat cancer-like symptoms. SG has been shown to have cytotoxic effects on cervical cancer cells (HeLa). Assays were done to identify the mechanisms of SG's cytotoxic effect. HeLa cells treated with SG exhibited early and late apoptosis, and caspase-9, -8 and -3 activities. It also induces generation of reactive oxygen species and disrupted mitochondrial membrane potential. SG isolated from A. fascicularis induces apoptosis through extrinsic and intrinsic pathways on HeLa cells. SG could be a candidate for in vivo studies and a promising natural compound in cervical cancer treatment.
    Keywords:  Secundiflorol G; apoptosis; cancer; cytotoxic
    DOI:  https://doi.org/10.1080/14786419.2019.1601092
  54. Mol Biol Rep. 2019 Apr 19.
    Zhu Y, Li Q, Xun W, Chen Y, Zhang C, Sun S.
      The purpose of our research is to elucidate whether oxLDL activates P2X7R in cultured human podocytes and if the activation of P2X7R leads to podocyte apoptosis. Additionally, we explore the underlying mechanism involved in podocyte apoptosis. Immortalized human podocytes were incubated with oxLDL (80 µg/ml), P2X7R antagonist A438079 (10 µM), or the compound of A438079 and oxLDL for 48 h, respectively. Cellular apoptosis and ROS were evaluated using flow cytometer. P2X7R, Bax, and Caspase-3 protein expression were detected by western blot and immunofluorescence analysis.The expression of P2X7R, ROS, Bax, and Caspase-3 in human podocytes incubated with oxLDL was significantly up-regulated and was found to have higher intracellular lipid accumulation and podocyte apoptosis compared with the NC group. However, co-administration with A438079, ROS, Bax, and Caspase-3 expression both significantly down-regulate as well as lower lipid accumulation and cellular apoptosis in the oxLDL-induced podocyte group. We revealed that P2X7R is involved in the regulation of oxLDL-treated podocytes. Additionally, we found that the anti-apoptotic effect of A438079 is correlated with ROS, Bax, and Caspase-3 expression down-regulated.
    Keywords:  Bax; Caspase-3; P2X7R; Podocyte; ROS; oxLDL
    DOI:  https://doi.org/10.1007/s11033-019-04823-6
  55. Front Cell Neurosci. 2019 ;13 123
    Deng X, Wang M, Hu S, Feng Y, Shao Y, Xie Y, Wu M, Chen Y, Shi X.
      Cognitive dysfunction is one of the serious complications induced by status epilepticus (SE), which has a significant negative impact on patients' quality of life. Previous studies demonstrated that the pathophysiological changes after SE such as oxidative stress, inflammatory reaction contribute to neuronal damage. A recent study indicated that preventive astaxanthin (AST) alleviated epilepsy-induced oxidative stress and neuronal apoptosis in the brain. In the present study, rats were treated with vehicle or AST 1 h after SE onset and were injected once every other day for 2 weeks (total of seven times). The results showed that the cognitive function in SE rats was significantly impaired, and AST treatment improved cognitive function in the Morris water maze (MWM). Magnetic resonance imaging (MRI), hematoxylin-eosin (HE) staining and TdT-mediated dUTP Nick-End Labeling (TUNEL) staining showed obvious damage in the hippocampus of SE rats, and AST alleviated the damage. Subsequently, we evaluated the effect of AST on relative pathophysiology to elucidate the possible mechanisms. To evaluate the oxidative stress, the expression of malondialdehyde (MDA) and superoxide dismutase (SOD) in plasma were detected using commercially available kits. NADPH oxidase-4 (Nox-4), p22phox, NF-E2-related factor 2 (Nrf-2), heme oxygenase 1 (Ho-1) and sod1 in the parahippocampal cortex and hippocampus were detected using western blot and real-time polymerase chain reaction (RT-PCR). The levels of MDA in plasma and Nox-4 and p22phox in the brain increased in SE rats, and the levels of SOD in plasma and Nrf-2, Ho-1 and sod1 in the brain decreased. Treatment with AST alleviated these changes. We also detected the levels of inflammatory mediators like cyclooxygenase-2 (cox-2), interleukin-1β (IL-1β), tumor necrosis factor-α (TNF-α) and NF-κB phosphorylation p65 (p-p65)/p65 in the brain. The inflammatory reaction was significantly activated in the brain of SE rats, and AST alleviated neuroinflammation. We detected the levels of p-Akt, Akt, B-cell lymphoma-2 (Bcl-2), Bax, cleaved caspase-3, and caspase-3 in the parahippocampal cortex and hippocampus using western blot. The levels of p-Akt/Akt and Bcl-2 decreased in SE rats, Bax and cleaved caspase-3/caspase-3 increased, while AST alleviated these changes. The present study indicated that AST exerted an reobvious neuroprotective effect in pilocarpine-induced SE rats.
    Keywords:  apoptosis; astaxanthin; neuroinflammation; neuroprotective effect; oxidative stress; status epilepticus
    DOI:  https://doi.org/10.3389/fncel.2019.00123
  56. Exp Ther Med. 2019 May;17(5): 3709-3716
    Yang C, Li B, Liu Y, Xing Y.
      Ginsenoside Rb1 (GS-Rb1) is one of the most important active pharmacological extracts of the traditional Chinese medicine, ginseng, and there is extensive evidence of its cardioprotective properties. However, the microRNA (miR) targets of GS-Rb1 and the underlying mechanisms of GS-Rb1 and miR-21 in the progression of cardiomyocyte apoptosis have not been clearly elucidated. The aim of the current study was to investigate the impact of miR-21 and its target gene, programmed cell death protein 4 (PDCD4), on the protective effect of GS-Rb1 in cardiomyocytes injured by oxygen-glucose deprivation (OGD). The miR-21 expression levels were downregulated, and the percentage of the apoptotic cells and reactive oxygen species (ROS) was increased in OGD-cultured neonatal rat cardiomyocytes; however, the effects were reversed by GS-Rb1 treatment. It was demonstrated that GS-Rb1 could reduce intracellular ROS content, and the expression of cytochrome C and the pro-apoptosis protein, apoptosis regulator B-cell lymphoma associated X (Bax) protein while increasing the expression of the anti-apoptosis protein, apoptosis regulator Bcl-2. The target gene, PDCD4, was significantly upregulated in the OGD group; however, the expression of PDCD4 was inhibited by GS-Rb1 treatment. Furthermore, miR-21 inhibitor transfection reduced GS-Rb1-induced miR-21 upregulation compared with the OGD+GS-Rb1 group, indicating that the miR-21 was involved in the anti-apoptotic effect of GS-Rb1 in cardiomyocytes. The results of the current study highlighted that GS-Rb1 could target miR-21 and its target gene, PDCD4, to protect OGD-injured cardiomyocytes. The results of the current study may provide a novel insight for the treatment of myocardial infarction with Traditional Chinese Medicines, involving miRs as targets.
    Keywords:  Ginsenoside Rb1; apoptosis; cardiomyocytes; microRNA-21; oxygen-glucose deprivation; programmed cell death protein 4
    DOI:  https://doi.org/10.3892/etm.2019.7330
  57. Hum Cell. 2019 Apr 16.
    Liu Y, Zhai J, Chen J, Wang X, Wen T.
      Obese women with polycystic ovary syndrome (PCOS) often suffer from ovulation failure, which may be driven by granulosa cells (GCs) injury caused by increased levels of circulating oxidized low-density lipoprotein (ox-LDL) and luteinizing hormone (LH). PGC-1α may play an important role in this pathophysiological processes. However, the effect and the potential mechanism of PGC-1α on GCs injury evoked by obese PCOS is fully unclear. To investigate the protective effect and the potential mechanism of PGC-1α on GCs injury evoked by ox-LDL + LH stimulation. Patients with PCOS and women of normal reproductive age who undergoing egg retrievals and consenting for this research were collected. Those women were divided into normal-weight non-PCOS group, obese non-PCOS group, normal-weight PCOS group and obese PCOS group according to the body mass index (BMI) and PCOS diagnosis. Follicular fluid was collected and primary GCs were isolated. The levels of LH and ox-LDL in follicular fluid in the four groups were measured. And, the expressions of PGC-1α, cell apoptosis and ROS generation in primary GCs in the four groups were evaluated. After GCs from women of normal reproductive age at normal-weight pre-treated with adenovirus encoding PGC-1α (Ad-PGC-1α) prior to ox-LDL + LH treatment in vitro, the cell viability, apoptosis, apoptosis-related proteins expressions and ROS generation were evaluated by CCK-8 assay, AnnexinV/PI double staining, Western blot and H2DCF-DA staining, respectively. The expression of PGC-1α was significantly decreased, whereas the cell apoptosis and ROS generation were significantly increased in GCs of PCOS group, especially obese PCOS group. Our data also revealed that over-expression of PGC-1α in GCs from women of normal reproductive age at normal-weight markedly inhibited cell injury, ROS generation and p38 activation, accompanied by increased Bcl-2 expression, decreased Bax and cleaved caspase-3 expressions induced by ox-LDL + LH stimulation. Ox-LDL + LH-induced cell apoptosis was abrogated by attenuation of ROS generation or p38 activation. Attenuation of ROS generation reversed ox-LDL + LH-induced p38 activation, however, p38 inhibitors had an effect on ROS generation. Our findings suggested that PGC-1α protected against ox-LDL + LH-induced GCs injury through inhibiting cell apoptosis. And, the mechanism may be related to the inhibition of ROS-initiated p38 pathway. Our data indicated that PGC-1α may be a potential therapeutic target for obese PCOS.
    Keywords:  Apoptosis; Granulosa cell; Oxidized low-density lipoprotein; PGC-1α; Polycystic ovary syndrome; ROS; p38
    DOI:  https://doi.org/10.1007/s13577-019-00252-6
  58. J Cell Biochem. 2019 Apr 16.
    Zhou W, Zhang S, Li J, Li Z, Wang Y, Li X.
      This study aims to investigate whether terminal differentiation-induced ncRNA (TINCR) has an effect on apoptosis and autophagy induced by ALA-PDT in cutaneous squamous cell carcinoma (CSCC). A431 cells were treated with 5-aminolevulinic acid (ALA) solution at different concentrations and for different duration time. A431 cell viability was detected by Cell Counting Kit-8 (CCK-8) assay, relative TINCR messenger RNA expression was detected by quantitative reverse-transcription polymerase chain reaction (qRT-PCR). A431 cell apoptosis was examined by flow cytometry. Relative apoptosis/autophagy-related protein expression was analyzed by Western blot analysis. The effect of TINCR on cell autophagy was detected by RFP-LC3 immunofluorescence assay. Reactive oxygen species concentration was detected by 2',7'-dichlorodihydrofluorescein diacetate (DCFH-DA) fluorescent probe. Relative expressions of ERK1/2 and specificity protein 3 (Sp3) in A43 cells were detected by Western blot analysis and qRT-PCR. Sp3 binding sites were analyzed by ChIP-qPCR. The relative transcription activity was measured with luciferase reporter assay. ALA-PDT treatment at 3.2 mmol/L for 120 minutes significantly promoted TINCR expression in CSCC A431 cells, and TINCR promoted ALA-PDT-induced apoptosis and cell autophagy. Furthermore, ALA-PDT promoted TINCR expression through ERK1/2-SP3 pathway. Sp3 promoted TINCR transcription by binding TINCR promoters. Our data indicated that TINCR involves in ALA-PDT-induced apoptosis and autophagy in CSCC.
    Keywords:  ERK1/2-Sp3 pathway; apoptosis; autophagy; terminal differentiation-induced ncRNA
    DOI:  https://doi.org/10.1002/jcb.28662
  59. Eur J Pharmacol. 2019 Apr 14. pii: S0014-2999(19)30249-3. [Epub ahead of print]
    Li C, Wang X, Cheng F, Du X, Yan J, Zhai C, Mu J, Wang Q.
      The blood-brain barrier (BBB) is involved in the pathogeneses of ischemic stroke (IS). Geniposide (GEN), an iridoid glycoside isolated from Gardenia jasminoides Ellis, has been used for the treatment of IS. However, the effects of GEN on the BBB are poorly understood. In vitro disease models of the BBB could be very helpful for the elucidation of underlying mechanisms and the development of novel therapeutic strategies. Therefore, we established an in vitro BBB model composed of primary cultures of brain microvascular endothelial cells and astrocytes. We then used this in vitro model to investigate the effect of GEN on the function of the BBB. Oxygen glucose deprivation and reoxygenation (OGD/R) significantly increased permeability and cell apoptosis in this in vitro BBB model. Notably, GEN pretreatment effectively improved the BBB function by decreasing the permeability of the BBB, promoting expression of tight junction proteins (zonula occludens-1, claudin-5, and occludin) and gamma-glutamyl transpeptidase, increasing transendothelial electrical resistance, mitigating oxidative stress damage and the release of inflammatory cytokines, downregulating the expression levels of matrix metallopeptidases-9 (MMP-9) and MMP-2, and increasing the release of brain derived neurotrophic factor and glial cell derived neurotrophic factor. Therefore, GEN can ameliorate the BBB dysfunction induced by OGD/R conditions through multiple protective mechanisms. The findings suggest that GEN may be an appropriate drug for restoring the barrier function of the BBB.
    Keywords:  Anti-apoptotic; Anti-inflammatory; Geniposide; In vitro blood-brain barrier; Oxygen glucose deprivation and reoxygenation; Tight junction
    DOI:  https://doi.org/10.1016/j.ejphar.2019.04.021
  60. Theriogenology. 2019 Apr 11. pii: S0093-691X(19)30092-5. [Epub ahead of print]132 72-82
    Zhang GM, An SY, El-Samahy MA, Zhang YL, Wan YJ, Wang ZY, Xiao SH, Meng FX, Wang F, Lei ZH.
      Peroxisome proliferator-activated receptor gamma coactivator-1 alpha (PPARGC1A) acts as a powerful coactivator of many transcriptional factors that relate to granulosa cell (GC) apoptosis. In this study, the miRNAs mediating goat follicular atresia and luteinized granulosa cell (LGC) apoptosis induced by hydrogen peroxide (H2O2) via PPARGC1A were investigated. Our results showed that miR-1197-3p targeted PPARGC1A was predicted by bioinformatics algorithm and verified by luciferase reporter assay. In addition, miR-1197-3p promoted goat LGC apoptosis via PPARGC1A through mitochondrial-dependent apoptosis pathway, and these effects could be restored by PPARGC1A overexpression. Moreover, H2O2-induced LGC apoptosis significantly upregulated miR-1197-3p expression and downregulated PPARGC1A level. Pretreatment of miR-1197-3p inhibitor alleviated LGC apoptosis induced by 400 μM H2O2 for 12 h, and preserved the mitochondrial membrane potential by increasing PPARGC1A expression. In conclusion, miR-1197-3p might act as an essential regulator of goat LGC apoptosis potentially via the mitochondrial-dependent apoptosis pathway by targeting PPARGC1A.
    Keywords:  Goat luteinized granulosa cells; Mitochondrial-dependent apoptosis pathway; PPARGC1A; miR-1197–3p
    DOI:  https://doi.org/10.1016/j.theriogenology.2019.04.008
  61. Toxins (Basel). 2019 Apr 16. pii: E226. [Epub ahead of print]11(4):
    Li H, Li S, Yang H, Wang Y, Wang J, Zheng N.
      The toxicity and related mechanisms of aflatoxin B1 (AFB1) and aflatoxin M1 (AFM1) in the mouse kidney were studied, and the role of l-proline in alleviating kidney damage was investigated. In a 28-day toxicity mouse model, thirty mice were divided into six groups: control (without treatment), l-proline group (10 g/kg body weight (b.w.)), AFB1 group (0.5 mg/kg b.w.), AFM1 (3.5 mg/kg b.w.), AFB1 + l-proline group and AFM1 + l-proline group. Kidney index and biochemical indicators were detected, and pathological staining was observed. Using a human embryonic kidney 293 (HEK 293) cell model, cell apoptosis rate and apoptotic proteins expressions were detected. The results showed that AFB1 and AFM1 activated pathways related with oxidative stress and caused kidney injury; l-proline significantly alleviated abnormal expressions of biochemical parameters and pathological kidney damage, as well as excessive cell apoptosis in the AF-treated models. Moreover, proline dehydrogenase (PRODH) was verified to regulate the levels of l-proline and downstream apoptotic factors (Bax, Bcl-2, and cleaved Caspase-3) compared with the control (p < 0.05). In conclusion, l-proline could protect mouse kidneys from AFB1 and AFM1 through alleviating oxidative damage and decreasing downstream apoptosis, which deserves further research and development.
    Keywords:  aflatoxin B1; aflatoxin M1; apoptosis; l-proline; proline dehydrogenase
    DOI:  https://doi.org/10.3390/toxins11040226
  62. Eur Rev Med Pharmacol Sci. 2019 Apr;pii: 17593. [Epub ahead of print]23(7): 3096-3104
    Fan S, Sun JB, Li R, Song X, Li J.
      OBJECTIVE: The aim of this research was to explore the protective effect of lycopene (Lyc) on myocardial ischemia injury through anti-apoptosis and anti-oxidative stress.MATERIALS AND METHODS: 75 rats were divided into 5 groups: sham operation group (control group), model group, low-dose group (Lyc+2 mg/kg), medium-dose group (Lyc+4 mg/kg) and high-dose group (Lyc+6 mg/kg). The rat model of myocardial ischemia was established by a subcutaneous injection of isoproterenol (85 mg/kg) for two consecutive days. Conventional HE staining and Masson staining were performed for pathological changes. Biochemical indicators were measured by the enzyme-linked immuno sorbent assay (ELISA). Western blotting was used to measure the levels of related proteins in JNK/STAT signaling pathway.
    RESULTS: Compared to control group, the levels of CK-MB, TC, and TGs were significantly increased in model group. The levels of CK-MB, TC, and TGs in each Lyc-administered group were decreased. After Lyc was administered, the SOD, CAT, GSH-Px activities and MDA content were all restored. The serum levels of IL-1β, TNF-α and IL-6 in control group were significantly lower than in model group. When the Lyc was administered, the serum IL-1β, TNF-α and IL-6 levels in medium-dose group and high-dose group were significantly decreased. The levels of Bax/Bcl-2, Cyt-c, and Caspase-3 in model group were significantly higher than control group. Changes of Bax/Bcl-2, Cyt-c, and Caspase-3 in medium-dose and high-dose groups after the administration of Lyc were restored significantly. The levels of p-JNK/JNK, p-STAT1 (Tyr701)/STAT1, p-STAT1 (Ser727)/STAT1, p-STAT3 (Tyr705)/STAT3 were significantly increased, while p-STAT3 (Ser727)/STAT3 was significantly decreased. When Lyc was administered, the expression levels of p-JAK/JAK, p-STAT1 (Tyr701)/STAT1, p-STAT1 (Ser727)/STAT1, p-STAT3 (Tyr705)/STAT3 protein in medium-dose group and high-dose group were significantly decreased, and the expression level of p-STAT3 (Ser727)/STAT3 protein was significantly increased.
    CONCLUSIONS: Lyc could show a protective effect on oxidative stress injury and anti-cardiomyocyte apoptosis of myocardial ischemia, and its possible mechanism was to attenuate the activation of JNK/ERK signaling pathway induced by myocardial injury.
    DOI:  https://doi.org/10.26355/eurrev_201904_17593
  63. Cancer Cell Int. 2019 ;19 81
    Xia Y, Liu X, Zhang L, Zhang J, Li C, Zhang N, Xu H, Li Y.
      Background: Gastric cancer, as a multifactorial disorders, shows cytological and architectural heterogeneity compared to other gastrointestinal cancers, making it therapeutically challenging. Cisplatin is generally used in clinic for gastric cancer treatment but with toxic side effects and develops resistance. Anti-tumor properties of copper and its coordinated compounds have been explored intensively in recent years.Methods: In this study, we synthesized a novel Schiff base copper coordinated compound (SBCCC) and examined its antitumor effects in two gastric cancer cell lines SGC-7901 and BGC-823 as well as a mouse model of gastric cancer.
    Results: The results show that SBCCC can significantly inhibit the proliferation of gastric cancer cells in a dose- and time-dependent manner. The IC50 of SBCCC in SGC-7901 and BGC-823 cells is 1 μM, which is much less than cisplatin's IC50. SBCCC induces apoptosis and causes cell cycle arrest at the G1 phase. SBCCC induces apoptosis via multiple pathways including inhibition of NF-κB, ROS production and autophagy.
    Conclusions: The synthesized SBCCC induced cancer cell death via inhibition of NF-κB, ROS production and autophagy. The multiple cell-killing mechanisms were important to overcome therapeutic failure because of multidrug-resistance of cancer cells. SBCCC, with a lower IC50 compared to cisplatin, could render it the potential to overcome the side-effect for clinical application.
    Keywords:  Autophagy; NF-κB; ROS; Schiff base copper coordination compound
    DOI:  https://doi.org/10.1186/s12935-019-0801-6
  64. Avian Pathol. 2019 Apr 15. 1-27
    Zhu T, Qi B, Liu R, Jiang X, Lu R, Xiao L, Fu G, Fu Q, Shi S, Wan C, Huang Y.
      Duck hepatitis A virus type 1 (DHAV-1) causes acute hepatitis with high morbidity and mortality in ducklings of the genera Cairina and Anas and is characterized by ecchymotic haemorrhage and necrosis of the liver surface. Since September 2011, a new subtype of DHAV-1 (named pancreatitis-type DHAV-1) has been isolated. This new subtype is characterized by yellowish or haemorrhagic pancreatitis, but with no significant pathological changes in the liver. To further investigate the difference in pathogenicity between hepatitis-type DHAV-1 and pancreatitis-type DHAV-1, we infected Muscovy ducklings with a hepatitis-type DHAV-1 strain, FZ86, and a pancreatitis-type DHAV-1 strain, MPZJ1206, respectively, and then compared the resulting gross lesions, histopathological changes, viral distribution and cellular apoptosis in the liver and pancreas of Muscovy ducklings. The results suggested that FZ86 induced a more efficient viral propagation in the liver than MPZJ1206, and the gross and histopathological lesions were also limited to the liver. However, MPZJ1206 induced more effective viral replication in the pancreas than FZ86. The MPZJ1206-infected Muscovy ducklings showed an obviously yellowed and haemorrhagic pancreas, but with no significant pathological changes in the liver. Furthermore, FZ86 induced notable hepatocyte apoptosis and increased the expression of caspase-3 in the liver, whereas MPZJ1206 caused apoptosis in a large number of acinar epithelial cells and elevated the expression of caspase-3 in the pancreas. Taken together, these results demonstrated that pancreatitis-type DHAV-1 has many new pathogenic features which distinguish it from the hepatitis-type DHAV-1. Research highlights Pancreatitis-type DHAV-1 (MPZJ1206) was characterized by pancreatic haemorrhage and yellow discoloration, but with no obvious haemorrhage and necrosis in the liver. Pancreatitis-type DHAV-1(MPZJ1206) exhibits many new pathogenic features which distinguish it from the hepatitis-type DHAV-1(FZ86).
    Keywords:  Duck hepatitis A virus type 1; apoptosis; duckling; liver; pancreas; pathogenicity
    DOI:  https://doi.org/10.1080/03079457.2019.1605146
  65. J Cell Physiol. 2019 Apr 19.
    Samimi M, Pourhanifeh MH, Mehdizadehkashi A, Eftekhar T, Asemi Z.
      Endometriosis is a frequent and chronic illness in young women which could be defined by the existence of endometrial stroma and glands outside of the normal site of the lining of the uterus. It has painful symptoms. The advanced stage of endometriosis may lead to gynecological malignancies, such as ovarian cancer, and other complications, including infertility. However, its exact physiopathology is not well known. Recent studies have shown the possible roles of inflammation along with oxidative stress. Additionally, angiogenesis and apoptosis dysregulation contribute to endometriosis pathophysiology. Therapeutic strategies and continuing attempts, to conquer endometriosis should be done regarding molecular signaling pathways. Thus, the present review summarizes current studies and focuses on molecular mechanisms.
    Keywords:  angiogenesis; apoptosis; endometriosis; inflammation; oxidative stress
    DOI:  https://doi.org/10.1002/jcp.28666
  66. J Cell Mol Med. 2019 Apr 15.
    Xia D, Zhai X, Wang H, Chen Z, Fu C, Zhu M.
      Alpha lipoic acid (ALA) is a powerful antioxidant which has been widely used in the treatment of different system diseases, such as cardiovascular and cerebrovascular diseases. But, there are few studies that refer to protective effects and potential mechanisms on traumatic brain injury (TBI). This study was carried out to investigate the neuroprotective effect following TBI and illuminate the underlying mechanism. Weight drop-injured model in rats was induced by weight-drop. ALA was administrated via intraperitoneal injection after TBI. Neurologic scores were examined following several tests. Neurological score was performed to measure behavioural outcomes. Nissl staining and TUNEL were performed to evaluate the neuronal apoptosis. Western blotting was engaged to analyse the protein content of the Nuclear factor erythroid 2-related factor 2 (Nrf2) and its downstream protein factors, including hemeoxygenase-1 (HO-1) and quinine oxidoreductase-1 (NQO1). ALA treatment alleviated TBI-induced neuron cell apoptosis and improved neurobehavioural function by up-regulation of Nrf2 expression and its downstream protein factors after TBI. This study presents new perspective of the mechanisms responsible for the neuronal apoptosis of ALA, with possible involvement of Nrf2 pathway.
    Keywords:  ALA; Nrf2; neuronal apoptosis; traumatic brain injury
    DOI:  https://doi.org/10.1111/jcmm.14296
  67. J Cell Physiol. 2019 Apr 16.
    Li M, Cui ZG, Zakki SA, Feng Q, Sun L, Feril LB, Inadera H.
      Chemoresistance is one of the major obstacles in chemotherapy-based hepatocellular carcinoma (HCC) intervention. Aluminum (Al) is an environmental pollutant that plays a vital role in carcinogenesis, tumorigenesis, and metastasis. However, the effect of Al on chemoresistance remains unknown. 5-Fluorouracil (5-FU) is a widely used antitumor drug. Therefore, we investigated the effects of aluminum chloride (AlCl3 ) on the chemoresistance of HepG2 cells to 5-FU and explored the underlying mechanisms of these effects. The results demonstrated that AlCl3 pretreatment attenuated 5-FU-induced apoptosis through Erk activation and reversed 5-FU-induced cell cycle arrest by downregulating p-Chk2Thr68 levels. In addition, AlCl3 markedly increased the levels of proteins associated with cell migration, such as MMP-2 and MMP-9. Further investigation demonstrated that an Erk inhibitor (U0126) reversed the AlCl3 -induced decrease in apoptosis, enhancement of cell cycle progression, promotion of cell migration, and attenuation of oxidative stress. In summary, AlCl3 induced chemoresistance to 5-FU in HepG2 cells. The present study suggests a potential influence of AlCl3 on 5-FU therapy. These findings may help others to understand and properly address the resistance of HCC to chemotherapeutic agents.
    Keywords:  5-FU; AlCl3; Erk signaling pathway; apoptosis; chemoresistance
    DOI:  https://doi.org/10.1002/jcp.28625
  68. Toxicol Sci. 2019 Apr 15. pii: kfz089. [Epub ahead of print]
    Jie G, Wang Y, Liu Y, Shi M, Yin L, Hou Y, Zhou Y, Wong CKC, Chen D, Guo Z, Shi H.
      Cadmium (Cd) is a toxic heavy metal that can accumulate and cause severe damage to many organs, such as liver, kidney, lung, etc. Cd also significantly suppresses immunity, however, the underlying mechanism involved in Cd-induced immunnotoxicity is still unclear. The present study indicated that semi-chronic Cd exposure (7 days) induced apoptotic damage of mouse spleen. In human Ramos B cells, Cd exposure also induced apoptosis, which was dependent on Cd induced vacuole membrane protein 1 (VMP1) expression and autophagy. Cd induced autophagy and apoptosis was abated when VMP1 expression was knockdown. In addition, Cd-induced VMP1 expression, autophagy and apoptosis were dependent on the elevation of Ca2+ and reactive oxygen species (ROS). More important, Cd exposure also induced VMP1 expression and autophagy in mouse spleen tissue, and the intraperitoneal injection of the autophagy inhibitor chloroquine (CQ) into mice effectively reduced Cd induced spleen apoptotic damage. Taken together, these results indicate Cd induced autophagy, promotes apoptosis in immune cells, and inhibition of autophagy can alleviate Cd induced spleen and immune cell apoptosis. This study might provide the groundwork for future studies on Cd induced immunomodulatory effects and immune diseases.
    Keywords:  Apoptosis; Autophagy; Cadmium; Immunotoxicity; VMP1
    DOI:  https://doi.org/10.1093/toxsci/kfz089
  69. ACS Appl Mater Interfaces. 2019 Apr 19.
    Lim WQ, Yang G, Phua SZF, Chen H, Zhao Y.
      Nanomedicine has emerged as a promising strategy for effective cancer treatment. A useful approach is to develop carrier-free nanodrugs via facile supramolecular self-assembly process. To achieve high therapeutic effect, integrating photodynamic therapy with chemotherapy has been sought after. In this work, we designed a nanocarrier (PEG-Por-CD: oxliPt(IV)-ada) assembled with oxaliplatin prodrug (oxliPt(IV)-ada) and porphyrin photosensitizer (PEG-Por-CD) through host-guest interaction to achieve stimulus-responsive combination therapy. Contributed by excellent spatial control of binding ratio between host and guest molecules, porphyrin and oxaliplatin were separately modified with β-cyclodextrin and adamantane to prepare the amphiphilic host-guest complex for subsequent self-assembly into therapeutic nanoparticles. The obtained PEG-Por-CD: oxliPt(IV)-ada nanoparticles exhibited good colloidal stability with an average hydrodynamic size of 164 nm, whilst undergoing the disassembly under reductive environment to release active therapeutic species. Confocal imaging demonstrated the ability of PEG-Por-CD: oxliPt(IV)-ada to effectively accumulate in the cells, and produce reactive oxygen species in vitro upon 630 nm light irradiation. As compared with the monotherapy, the PEG-Por-CD: oxliPt(IV)-ada nanoparticles exhibited 3-folds enhanced cytotoxicity and 2-folds increase in the apoptosis. In vivo experiments using 4T1 tumor-bearing mice confirmed that the nanoparticles were efficient in suppressing the tumor growth without eliciting systemic toxicity. The present self-delivery nanosystem constructed from the self-assembly approach not only allows precise control over the drug and photosensitizer loading ratio, but also eliminates systemic toxicity concern of the drug carriers, providing a solution for further development of combinational cancer treatment.
    DOI:  https://doi.org/10.1021/acsami.9b04557
  70. Daru. 2019 Apr 18.
    Srivastava S, Mishra S, Dewangan J, Divakar A, Gupta N, Kalleti N, Mugale MN, Kumar S, Sharma S, Rath SK.
      BACKGROUND: Diethylene glycol monoethyl ether (DEGEE) is widely used as a solubilizer in cosmetics as well as in oral, topical, transdermal and injectable pharmaceutical formulations. Due to the unavailability of detailed toxicological studies on DEGEE, the Scientific Committee on Consumer Products (SCCP) found its toxicological reports to be unsatisfactory, comprising only summaries. Also, a few reports have raised concern on the use of DEGEE as it might cause damage to the kidneys.OBJECTIVE: Safety assessment of DEGEE using in vitro and in vivo models.
    METHODS: In vitro effects of DEGEE (0.5-25 mg/ml) were assessed in the HEK293 human embryonic kidney cells. In vivo effects were evaluated after single acute exposure of DEGEE via intraperitoneal route in Swiss albino mice and further, a 28 days subchronic exposure study was conducted where DEGEE was administered orally, once daily.
    RESULTS: DEGEE was cytotoxic to HEK293 cells, and an IC50 of 15 mg/ml was established. An increase in the intracellular levels of ROS and alteration in the mitochondrial membrane potential led to nuclear fragmentation and induction of apoptosis in these cells. Survival rate of animals administered intraperitoneally with a single acute dose of 1000 mg/kg DEGEE was 100% with no significant changes in the behavioural and histological parameters. However, the dose of 3000 mg/kg and above led to total mortality within 14 days of acute exposure. Subchronic oral exposure of 500-2000 mg/kg DEGEE showed no significant changes in the hematological, biochemical and histopathological parameters.
    CONCLUSIONS: The in vitro findings indicate that the nephrotoxic potential of DEGEE cannot be ruled out. The results of the in vivo studies reveal that the degree of toxic effects shown by DEGEE varies, depending on the dose, duration of exposure and routes of administration. Therefore, the present findings are of relevance and thorough studies should be conducted before using this substance in clinical formulations. Graphical abstract Evaluation of the toxic potential of Diethylene glycol monoethyl ether.
    Keywords:  Acute exposure; Diethylene glycol monoethyl ether; Excipient; Safety assessment; Subchronic exposure
    DOI:  https://doi.org/10.1007/s40199-019-00264-5
  71. Mar Drugs. 2019 Apr 02. pii: E207. [Epub ahead of print]17(4):
    Xu K, Guo C, Shi D, Meng J, Tian H, Guo S.
      A study on the secondary metabolites of Aspergillus sp. XNM-4, which was derived from marine algae Leathesia nana (Chordariaceae), led to the identification of one previously undescribed (1) and seventeen known compounds (2-18). Their planar structures were established by extensive spectroscopic analyses, while the stereochemical assignments were defined by electronic circular dichroism (ECD) calculations. The biological activities of the compounds were assessed on five human cancer cell lines (PANC-1, A549, MDA-MB-231, Caco-2, and SK-OV-3), and one human normal cell line (HL-7702) using an MTT [3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl tetrazolium bromide] assay. Among them, the dimeric naphthopyrones 7, 10 and 12 exhibited potent cytotoxicity. Further mechanism studies showed that 12 induced apoptosis, arrested the cell cycle at the G0/G1 phase in the PANC-1 cells, caused morphological changes and generated ROS; and it induces PANC-1 cells apoptosis via ROS-mediated PI3K/Akt signaling pathway.
    Keywords:  Aspergillus; Leathesia nana; cytotoxicity; endophytic fungus; naphthopyrones
    DOI:  https://doi.org/10.3390/md17040207