bims-aporos Biomed news
on Apoptosis and reactive oxygen species
Issue of 2019‒03‒31
seventy-one papers selected by
Gavin McStay
Staffordshire University


  1. Int J Environ Res Public Health. 2019 Mar 22. pii: E1044. [Epub ahead of print]16(6):
    Park C, Kwon DH, Hwang SJ, Han MH, Jeong JW, Hong SH, Cha HJ, Hong SH, Kim GY, Lee HJ, Kim S, Kim HS, Choi YH.
      Tacrolimus is widely used as an immunosuppressant to reduce the risk of rejection after organ transplantation, but its cytotoxicity is problematic. Nargenicin A1 is an antibiotic extracted from Nocardia argentinensis and is known to have antioxidant activity, though its mode of action is unknown. The present study was undertaken to evaluate the protective effects of nargenicin A1 on DNA damage and apoptosis induced by tacrolimus in hirame natural embryo (HINAE) cells. We found that reduced HINAE cell survival by tacrolimus was due to the induction of DNA damage and apoptosis, both of which were prevented by co-treating nargenicin A1 or N-acetyl-l-cysteine, a reactive oxygen species (ROS) scavenger, with tacrolimus. In addition, apoptosis induction by tacrolimus was accompanied by increases in ROS generation and decreases in adenosine triphosphate (ATP) levels caused by mitochondrial dysfunction, and these changes were significantly attenuated in the presence of nargenicin A1, which further indicated tacrolimus-induced apoptosis involved an oxidative stress-associated mechanism. Furthermore, nargenicin A1 suppressed tacrolimus-induced B-cell lymphoma-2 (Bcl-2) down-regulation, Bax up-regulation, and caspase-3 activation. Collectively, these results demonstrate that nargenicin A1 protects HINAE cells against tacrolimus-induced DNA damage and apoptosis, at least in part, by scavenging ROS and thus suppressing the mitochondrial-dependent apoptotic pathway.
    Keywords:  DNA damage; ROS; apoptosis; nargenicin A1; tacrolimus
    DOI:  https://doi.org/10.3390/ijerph16061044
  2. Food Chem Toxicol. 2019 Mar 21. pii: S0278-6915(19)30157-7. [Epub ahead of print]127 188-196
    Hu T, Linghu K, Huang S, Battino M, Georgiev MI, Zengin G, Li D, Deng Y, Wang YT, Cao H.
      Significant evidence indicated that flaxseed (Linum usitatissimum) possesses various positive health aspects such as reducing the risk of cancer and cardiovascular diseases. The fatty acids are considered to be responsible for these benefits of flaxseed. Herein, the in vitro effects of flaxseed extract on the growth and apoptosis of human breast cancer MCF-7 cells were investigated. The MCF-7 cells treated with flaxseed extract showed a dose-dependent decrease in cell viability. The flaxseed extract induced reactive oxygen species and the flow cytometric analysis demonstrated that flaxseed fatty acids triggered apoptosis of MCF-7 cells, which was also shown by the loss of mitochondrial membrane potential and caspase cascade reaction. Thus, the flaxseed extract regulated the growth of MCF-7 cells and induced apoptosis. Eventually, the flaxseed could be used as a dietary supplement to prevent breast cancer.
    Keywords:  Apoptosis; Dietary supplement; Flaxseed; MCF-7 cells; Mitochondrial membrane potential; ROS
    DOI:  https://doi.org/10.1016/j.fct.2019.03.029
  3. Oxid Med Cell Longev. 2019 ;2019 7192798
    Liu ZJ, Zhao W, Lei HY, Xu HL, Lai LY, Xu R, Xu SY.
      Bupivacaine, a typical local anesthetic, induces neurotoxicity via reactive oxygen species regulation of apoptosis. High glucose could enhance bupivacaine-induced neurotoxicity through regulating oxidative stress, but the mechanism of it is not clear. Mitochondrial calcium uniporter (MCU), a key channel for regulating the mitochondrial Ca2+ (mCa2+) influx, is closely related to oxidative stress via disruption of mCa2+ homeostasis. Whether MCU is involved in high glucose-sensitized bupivacaine-induced neurotoxicity remains unknown. In this study, human neuroblastoma (SH-SY5Y) cells were cultured with high glucose and/or bupivacaine, and the data showed that high glucose enhanced bupivacaine-induced MCU expression elevation, mCa2+ accumulation, and oxidative damage. Next, Ru360, an inhibitor of MCU, was employed to pretreated SH-SY5Y cells, and the results showed that it could decrease high glucose and bupivacaine-induced mCa2+ accumulation, oxidative stress, and apoptosis. Further, with the knockdown of MCU with a specific small interfering RNA (siRNA) in SH-SY5Y cells, we found that it also could inhibit high glucose and bupivacaine-induced mCa2+ accumulation, oxidative stress, and apoptosis. We propose that downregulation expression or activity inhibition of the MCU channel might be useful for restoring the mitochondrial function and combating high glucose and bupivacaine-induced neurotoxicity. In conclusion, our study demonstrated the crucial role of MCU in high glucose-mediated enhancement of bupivacaine-induced neurotoxicity, suggesting the possible use of this channel as a target for curing bupivacaine-induced neurotoxicity in diabetic patients.
    DOI:  https://doi.org/10.1155/2019/7192798
  4. Toxicol Lett. 2019 Mar 22. pii: S0378-4274(18)31775-2. [Epub ahead of print]
    Zhang G, Yang W, Jiang F, Zou P, Zeng Y, Ling X, Zhou Z, Cao J, Ao L.
      Dibutyl phthalate (DBP)-induced germ cell apoptosis contributes to male reproductive toxicity, however, the primary target organelle of DBP or the molecular events triggered by DBP to initiate germ cell apoptosis remain unclear. Our previous studies demonstrated DBP could stimulate the production of intracellular reactive oxygen species (ROS), which served as an upstream mediator of activation of endoplasmic reticulum (ER) stress in mouse spermatocyte-derived GC-2 cells. In the present study, the impacts of DBP-induced ROS generation on the mitochondria-related damage and the associations between ER stress and mitochondrial-related damage were investigated in GC-2 cells. We observed significant decreases of mitochondrial mass, mtDNA copy number, COX IV protein level, and ATP level in DBP-treated GC-2 cells in a dose-dependent manner. And DBP activated mitochondrial-related apoptosis, indicated by the elevation of cytoplasmic cytochrome C (Cyt C) and the activation of caspase-9/3 cascade. Pretreatment with antioxidant melatonin obviously attenuated DBP-induced mitochondrial damage and mitochondrial-dependent apoptosis in GC-2 cells, indicating the role of ROS in DBP-caused testicular toxicity. In response to oxidative stress, the Nrf2/ARE axis was activated in DBP-treated GC-2 cells to counteract ROS overproduction and subsequent mitochondrial damage. Further experiments showed DBP treatment increased the phosphorylated expression of ER stress-related protein PERK. GSK2606414, a specific inhibitor of PERK, partly attenuated the expression of Nrf2. And both DBP-induced mitochondrial damage in GC-2 cells and mitochondrial-dependent apoptosis of the germ cells in rat testes were further aggravated by PERK inhibition. Taken together, our data suggest that PERK regulates the Nrf2/ARE antioxidant pathway functioning as a self-defense mechanism against ROS-related mitochondrial damage induced by DBP in male germ cells.
    Keywords:  Dibutyl phthalate; Mitochondrial damage; Nrf2/ARE axis; PERK; Reactive oxygen species
    DOI:  https://doi.org/10.1016/j.toxlet.2019.03.007
  5. Food Chem Toxicol. 2019 Mar 22. pii: S0278-6915(19)30162-0. [Epub ahead of print]
    Ahmad A, Zafar A, Ahmad M.
      Edifenphos (EDF) is an organophosphorous pesticide and used in agriculture for pest control. However, EDF has been shown to accumulate in agricultural products and causes hazards to human health. Although reports are available regarding environmental impact of EDF, toxic effects of EDF on human cellular system especially immune cells have not been elucidated. In this study, genotoxicity and cytotoxicity of EDF on human peripheral blood lymphocytes and its amelioration by apigenin (dietary flavonoid) was investigated. We demonstrated that EDF inhibited cell viability, and induced oxidative stress and DNA damage in lymphocytes. In addition, results indicate that EDF induced apoptosis in lymphocytes concurrent with ROS generation, loss of mitochondrial membrane potential, up-regulation of Bax and caspase-9/-3 activation. Mechanistically, incubation of lymphocytes with N-acetylcysteine (ROS scavenger) abrogated the ROS generation and apoptosis caused by EDF. These findings suggest that ROS generation by EDF acts as an upstream signal leading to DNA damage and apoptosis in lymphocytes. This study also showed that apigenin could potentially attenuate EDF-induced oxidative stress, DNA damage and apoptosis in lymphocytes. Collectively, these results suggest that EDF exerts cytotoxicity and DNA damage in lymphocytes, and apigenin could be a potent dietary anti-oxidant regimen against EDF-induced toxicity on human health.
    Keywords:  Apigenin; Apoptosis; DNA damage; Edifenphos; Lymphocytes; Toxicity
    DOI:  https://doi.org/10.1016/j.fct.2019.03.034
  6. Drug Dev Res. 2019 Mar 27.
    Zang YQ, Feng YY, Luo YH, Zhai YQ, Ju XY, Feng YC, Wang JR, Yu CQ, Jin CH.
      Glycitein is an isoflavone that reportedly inhibits the proliferation of human breast cancer and prostate cancer cells. However, its anti-cancer molecular mechanisms in human gastric cancer remain to be defined. This study evaluated the antitumor effects of glycitein on human gastric cancer cells and investigated the underlying mechanisms. We used MTT assay, flow cytometry and western blotting to investigate its molecular mechanisms with focus on reactive oxygen species (ROS) production. Our results showed that glycitein had significant cytotoxic effects on human gastric cancer cells. Glycitein markedly decreased mitochondrial transmembrane potential (ΔΨm) and increased AGS cells mitochondrial-related apoptosis, and caused G0/G1 cell cycle arrest by regulating cycle-related protein. Mechanistically, accompanying ROS, glycitein can activate mitogen-activated protein kinase (MAPK) and inhibited the signal transducer and activator of transcription 3 (STAT3) and nuclear factor-kappaB (NF-κB) signaling pathways. Furthermore, the MAPK signaling pathway regulated the expression levels of STAT3 and NF-κB upon treatment with MAPK inhibitor and N-acetyl-L-cysteine (NAC). These findings suggested that glycitein induced AGS cell apoptosis and G0/G1 phase cell cycle arrest via ROS-related MAPK/STAT3/NF-κB signaling pathways. Thus, glycitein has the potential to a novel targeted therapeutic agent for human gastric cancer.
    Keywords:  apoptosis; cell cycle arrest; glycitein; human gastric cancer; signaling pathways
    DOI:  https://doi.org/10.1002/ddr.21534
  7. Chem Biol Interact. 2019 Mar 25. pii: S0009-2797(18)31051-2. [Epub ahead of print]
    Nguyen PT, Dang DK, Tran HQ, Shin EJ, Jeong JH, Nah SY, Chang Cho M, Lee YS, Jang CG, Kim HC.
      Methiopropamine (MPA) is structurally categorized as a thiophene ring-based methamphetamine (MA) derivative. Although abusive potential of MPA was recognized, little is known about the neurotoxic potential of MPA up to now. We investigated whether MPA induces dopaminergic neurotoxicity, and whether MPA activates a specific dopamine receptor. Here, we observed that treatment with MPA resulted in dopaminergic neurotoxicity in a dose-dependent manner. MPA treatment potentiated oxidative parameters (i.e., increases in the level of reactive oxygen species, 4-hydroxynonenal, and protein carbonyl), M1 phenotype-related microglial activity, and pro-apoptotic property (i.e., increases in Bax- and cleaved caspase-3-expressions, while a decrease in Bcl-2-expression). Moreover, treatment with MPA resulted in significant impairments in dopaminergic parameters [i.e., changes in dopamine level, dopamine turnover rate, tyrosine hydroxylase (TH) levels, dopamine transporter (DAT) expression, and vesicular monoamine transporter-2 (VMAT-2) expression], and in behavioral deficits. Both dopamine D1 receptor antagonist SCH23390 and D2 receptor antagonist sulpiride protected from these neurotoxic consequences. Therefore, our results suggest that dopamine D1 and D2 receptors simultaneously mediate MPA-induced dopaminergic neurodegeneration in mice via oxidative burdens, microgliosis, and pro-apoptosis.
    Keywords:  Apoptosis; Dopamine receptors; Dopaminergic deficits; Methiopropamine; Microgliosis; Oxidative stress
    DOI:  https://doi.org/10.1016/j.cbi.2019.03.017
  8. J Cell Physiol. 2019 Mar 25.
    Zhai Y, Behera J, Tyagi SC, Tyagi N.
      Homocysteine (Hcy) is detrimental to bone health in a mouse model of diet-induced hyperhomocysteinemia (HHcy). However, little is known about Hcy-mediated osteoblast dysfunction via mitochondrial oxidative damage. Hydrogen sulfide (H2 S) has potent antioxidant, anti-inflammatory, and antiapoptotic effects. In this study, we hypothesized that the H2 S mediated recovery of osteoblast dysfunction by maintaining mitochondrial biogenesis in Hcy-treated osteoblast cultures in vitro. MC3T3-E1 osteoblastic cells were exposed to Hcy treatment in the presence or absence of an H2 S donor (NaHS). Cell viability, osteogenic differentiation, reactive oxygen species (ROS) production were determined. Mitochondrial DNA copy number, adenosine triphosphate (ATP) production, and oxygen consumption were also measured. Our results demonstrated that administration of Hcy increases the intracellular Hcy level and decreases intracellular H2 S level and expression of the cystathionine β-synthase/Cystathionine γ-lyase system, thereby inhibiting osteogenic differentiation. Pretreatment with NaHS attenuated Hcy-induced mitochondrial toxicity (production of total ROS and mito-ROS, ratio of mitochondrial fission (DRP-1)/fusion (Mfn-2)) and restored ATP production and mitochondrial DNA copy numbers as well as oxygen consumption in the osteoblast as compared with the control, indicating its protective effects against Hcy-induced mitochondrial toxicity. In addition, NaHS also decreased the release of cytochrome c from the mitochondria to the cytosol, which induces cell apoptosis. Finally, flow cytometry confirmed that NaHS can rescue cells from apoptosis induced by Hcy. Our studies strongly suggest that NaHS has beneficial effects on mitochondrial toxicity, and could be developed as a potential therapeutic agent against HHcy-induced mitochondrial dysfunction in cultured osteoblasts in vitro.
    Keywords:  hydrogen sulfide; methionine; mitochondria; osteoblast; oxidative stress
    DOI:  https://doi.org/10.1002/jcp.28498
  9. Cancers (Basel). 2019 Mar 26. pii: E430. [Epub ahead of print]11(3):
    Gualdani R, de Clippele M, Ratbi I, Gailly P, Tajeddine N.
      Cisplatin (CDDP) is one of the principal chemotherapeutic agents used for the first-line treatment of many malignancies, including non-small cell lung carcinoma (NSCLC). Despite its use for over 40 years, its mechanism of action is not yet fully understood. Store-operated calcium entry (SOCE), the main pathway allowing Ca2+ entry in non-excitable cells, is involved in tumorogenesis, cancer progression and chemoresistance. It has become an attractive target in cancer treatment. In this study, we showed that siRNA-mediated depletion of stromal interaction molecule 1 (STIM1) and transient receptor potential channel 1 (TRPC1), two players of the store-operated calcium entry, dramatically reduced CDDP cytotoxicity in NSCLC cells. This was associated with an inhibition of the DNA damage response (DDR) triggered by CDDP. Moreover, STIM1 depletion also reduced CDDP-dependent oxidative stress. In parallel, SOCE activation induced Ca2+ entry into the mitochondria, a major source of reactive oxygen species (ROS) within the cell. This effect was highly decreased in STIM1-depleted cells. We then conclude that mitochondrial Ca2+ peak associated to the SOCE contributes to CDDP-induced ROS production, DDR and subsequent apoptosis. To the best of our knowledge, this is the first time that it is shown that Ca2+ signalling constitutes an initial step in CDDP-induced apoptosis.
    Keywords:  apoptosis; cisplatin; mitochondrial calcium; non-small cell lung carcinoma; reactive oxygen species; store-operated calcium entry
    DOI:  https://doi.org/10.3390/cancers11030430
  10. Oxid Med Cell Longev. 2019 ;2019 4565238
    Vázquez-Martínez O, Valente-Godínez H, Quintanar-Stephano A, Gasca-Martínez D, López-Cervantes ML, Palma-Tirado L, de Jesús Guerrero-Carrillo M, Pérez-Solís M, Díaz-Muñoz M.
      A surgical connection between portal and inferior cava veins was performed to generate an experimental model of high circulating ammonium and hepatic hypofunctioning. After 13 weeks of portacaval anastomosis (PCA), hyperammonemia and shrinkage in the liver were observed. Low glycemic levels accompanied by elevated levels of serum alanine aminotransferase were recorded. However, the activity of serum aspartate aminotransferase was reduced, without change in circulating urea. Histological and ultrastructural observations revealed ongoing vascularization and alterations in the hepatocyte nucleus (reduced diameter with indentations), fewer mitochondria, and numerous ribosomes in the endoplasmic reticulum. High activity of hepatic caspase-3 suggested apoptosis. PCA promoted a marked reduction in lipid peroxidation determined by TBARs in liver homogenate but specially in the mitochondrial and microsomal fractions. The reduced lipoperoxidative activity was also detected in assays supplemented with Fe2+. Only discreet changes were observed in conjugated dienes. Fluorescent probes showed significant attenuation in mitochondrial membrane potential, reactive oxygen species (ROS), and calcium content. Rats with PCA also showed reduced food intake and decreased energy expenditure through indirect calorimetry by measuring oxygen consumption with an open-flow respirometric system. We conclude that experimental PCA promotes an angiogenic state in the liver to confront the altered blood flow by reducing the prooxidant reactions associated with lower metabolic rate, along with significant reduction of mitochondrial content, but without a clear hepatic dysfunction.
    DOI:  https://doi.org/10.1155/2019/4565238
  11. J Appl Toxicol. 2019 Mar 25.
    Gao H, Chen J, Ding F, Chou X, Zhang X, Wan Y, Hu J, Wu Q.
      Glyphosate-based herbicides have been used worldwide for decades and have been suggested to induce nephrotoxicity, but the underlying mechanism is not yet clear. In this study, we treated a human renal proximal tubule cell line (HK-2) with glyphosate for 24 hours at concentrations of 0, 20, 40 and 60 μm. Glyphosate was found to reduce cell viability and induce apoptosis and oxidative stress in a dose-dependent manner. Because the chemical structures of glyphosate and those of its metabolite AMPA are similar to glycine and glutamate, which are agonists of the N-methyl-d-aspartate receptor (NMDAR), we investigated the potential role of the NMDAR pathway in mediating the proapoptotic effect of glyphosate on proximal tubule cells. We found that NMDAR1 expression, as well as intracellular Ca2+ ([Ca2+ ]i ) and reactive oxygen species (ROS) levels, increased after glyphosate treatment. Blocking NMDAR attenuated glyphosate-induced upregulation of [Ca2+ ]i and ROS levels as well as apoptosis. Meanwhile, inhibition of [Ca2+ ]i reduced glyphosate-induced ROS and apoptosis, and inhibition of ROS alleviated glyphosate-induced apoptosis. In mice exposed to 400 mg/kg glyphosate, the urine low molecular weight protein levels started to increase from day 7. Upregulation of apoptosis and NMDAR1 expression in renal proximal tubule epithelium and an imbalance of oxidant and antioxidative products were observed. These results strongly suggest that activation of the NMDAR1 pathway, together with its downstream [Ca2+ ]i and oxidative stress, is involved in glyphosate-induced renal proximal tubule epithelium apoptosis.
    Keywords:  NMDA receptor; apoptosis; glyphosate; kidney; oxidative stress
    DOI:  https://doi.org/10.1002/jat.3795
  12. Chin Med. 2019 ;14 7
    Wang Z, Yin J, Li M, Shen J, Xiao Z, Zhao Y, Huang C, Zhang H, Zhang Z, Cho CH, Wu X.
      Background: Shikonin (SKN), a naphthoquinone compound, is isolated from Chinese herbal medicine Lithospermum root and has been studied as an anticancer drug candidate in human tumor models. This study is designed to investigate whether SKN can sensitize the therapeutic effect of paclitaxel (PTX) in drug-resistant human ovarian carcinoma cells.Methods: Human ovarian carcinoma A2780 cell along with the paired PTX-resistant A2780/PTX cells were used. The effects of SKN, PTX or their combination on cell viability were conducted using Sulforhodamine B assay. P-glycoprotein (P-gp) expression was analyzed by flow cytometry after staining with P-gp-FITC anti-body. P-gp activity was determined by a fluorometric MDR assay kit or a rhodamine 123-based efflux assay, respectively. Apoptosis was evaluated by flow cytometry after Annexin V-FITC/PI co-staining. The effect of SKN, PTX or their combination on reactive oxygen species (ROS) generation and expression of pyruvate kinase M2 (PKM2) were investigated using flow cytometry or western blotting, respectively. PKM2 activity was detected by a Pyruvate Kinase Assay Kit.
    Results: SKN/PTX co-treatment led to synergistically enhanced cytotoxicity and apoptosis in PTX-resistant ovarian cancer cells, indicating the circumvention of multidrug resistance (MDR) of PTX by SKN. Further study indicated that the MDR reversal effect of SKN was independent of inhibiting activity of the efflux transporter P-gp. Notably, SKN/PTX significantly increased the generation of intracellular ROS in A2780/PTX cells, and scavenging intracellular ROS blocked the sensitizing effects of SKN in PTX-induced cytotoxicity and apoptosis in A2780/PTX cells, but not in A2780 cells. Furthermore, SKN/PTX-induced downregulation of PKM2 (a key enzyme in glycolysis) and the suppression of its activity were inhibited by a ROS scavenger N-acetyl cysteine (NAC), suggesting that the synergy of the SKN/PTX combination may be not rely on PKM2 suppression.
    Conclusions: These results reveal a P-gp-independent mechanism through ROS generation for the SKN/PTX combination to overcome MDR in ovarian cancer.
    Keywords:  Multidrug resistance; P-glycoprotein; Paclitaxel; Pyruvate kinase M2; Reactive oxygen species; Shikonin
    DOI:  https://doi.org/10.1186/s13020-019-0231-3
  13. Nanomedicine (Lond). 2019 Mar 27.
    Buz PT, Duman FD, Erkisa M, Demirci G, Ari F, Ulukaya E, Acar HY.
      AIM: N-acetyl-L-cysteine (NAC) is a free radical scavenger. We developed NAC-coated Ag2S (NAC-Ag2S) quantum dot (QD) as an optical imaging and therapeutic agent.MATERIALS & METHODS: QDs were synthesized in water. Their optical imaging potential and toxicity were studied in vitro.
    RESULTS: NAC-Ag2S QDs have strong emission, that is tunable between 748 and 840 nm, and are stable in biologically relevant media. QDs showed significant differences both in cell internalization and toxicity in vitro. QDs were quite toxic to breast and cervical cancer cells but not to lung derived cells despite the higher uptake. NAC-Ag2S reduces reactive oxygen species (ROS) but causes cell death via DNA damage and apoptosis.
    CONCLUSION: NAC-Ag2S QDs are stable and strong signal-generating theranostic agents offering selective therapeutic effects.
    Keywords:  AgS quantum dots; DNA damage; N-acetyl-L-cysteine (NAC); apoptosis; imaging; reactive oxygen species (ROS)
    DOI:  https://doi.org/10.2217/nnm-2018-0214
  14. Hum Exp Toxicol. 2019 Mar 29. 960327119839173
    Gheena S, Ezhilarasan D.
      Hepatocellular carcinoma is the second most common cause of cancer death in the world and its incidence has dramatically increased worldwide in the past two decades. Syringic acid (SA) has been studied for its hepatoprotective, anti-inflammatory, immunomodulatory, free radical scavenging, and antioxidant activities. We aimed to evaluate the cytotoxic effect of SA against human hepatoma HepG2 cell line. Cytotoxicity was evaluated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. HepG2 cells were treated with SA at concentration ranges of 25, 50, and 100 µM for 24 h. Reactive oxygen species (ROS) expression was investigated by dichlorofluorescein staining assay. Morphological changes of SA-treated HepG2 cells were evaluated by acridine orange (AO) and ethidium bromide (EB) dual staining. Apoptotic marker gene expressions were evaluated by qPCR. SA treatment caused significant cytotoxicity and liberation of ROS in HepG2 cells. AO and EB staining showed membrane blebbing and distortion in SA-treated cells. Apoptotic markers such as caspases 3 and 9, cytochrome c, Apaf-1, Bax, and p53 gene expressions were significantly increased upon SA treatment indicating the possibility of apoptosis induction in HepG2 cells. This treatment also caused significant downregulation of Bcl-2 gene expression. SA has a cytotoxic effect on human HepG2 cell line, and this might be a promising agent in anticancer research.
    Keywords:  HepG2; Syringic acid; apoptosis; cytotoxicity; hepatocellular carcinoma
    DOI:  https://doi.org/10.1177/0960327119839173
  15. BMC Vet Res. 2019 Mar 25. 15(1): 98
    Ye C, Li R, Xu L, Qiu Y, Fu S, Liu Y, Wu Z, Hou Y, Hu CA.
      BACKGROUND: Haemophilus parasuis (HPS) is the causative agent of Glässer's disease, characterized by arthritis, fibrinous polyserositis and meningitis, and resulting in worldwide economic losses in the swine industry. Baicalin (BA), a commonly used traditional Chinese medication, has been shown to possess a series of activities, such as anti-bacterial, anti-viral, anti-tumor, anti-oxidant and anti-inflammatory activities. However, whether BA has anti-apoptotic effects following HPS infection is unclear. Here, we investigated the anti-apoptotic effects and mechanisms of BA in HPS-induced apoptosis via the protein kinase C (PKC)-mitogen-activated protein kinase (MAPK) pathway in piglet's mononuclear phagocytes (PMNP).RESULTS: Our data demonstrated that HPS could induce reactive oxygen species (ROS) production, arrest the cell cycle and promote apoptosis via the PKC-MAPK signaling pathway in PMNP. Moreover, when BA was administered, we observed a reduction in ROS production, suppression of cleavage of caspase-3 in inducing apoptosis, and inhibition of activation of the PKC-MAPK signaling pathway for down-regulating p-JNK, p-p38, p-ERK, p-PKC-α and PKC-δ in PMNP triggered by HPS.
    CONCLUSIONS: Our data strongly suggest that BA can reverse the apoptosis initiated by HPS through regulating the PKC-MAPK signaling pathway, which represents a promising therapeutic agent in the treatment of HPS infection.
    Keywords:  Apoptosis; Baicalin; Glässer’s disease; Haemophilus parasuis; PKC–MAPK
    DOI:  https://doi.org/10.1186/s12917-019-1840-x
  16. Int J Mol Sci. 2019 Mar 26. pii: E1508. [Epub ahead of print]20(6):
    Lee CM, Lee J, Nam MJ, Choi YS, Park SH.
      Tomentosin is a natural sesquiterpene lactone extracted from various plants and is widely used as a medicine because it exhibits essential therapeutic properties. In this study, we investigated the anti-carcinogenic effects of tomentosin in human osteosarcoma MG-63 cells by performing cell migration/viability/proliferation, apoptosis, and reactive oxygen species (ROS) analysis assays. MG-63 cells were treated with various doses of tomentosin. After treatment with tomentosin, MG-63 cells were analyzed using the MTT assay, colony formation assay, cell counting assay, wound healing assay, Boyden chamber assay, zymography assay, cell cycle analysis, FITC Annexin V apoptosis assay, terminal deoxynucleotidyl transferase dUTP nick end labeling assay, western blot analysis, and ROS detection analysis. Our results indicated that tomentosin decreased cell viability and migration ability in MG-63 cells. Moreover, tomentosin induced apoptosis, cell cycle arrest, DNA damage, and ROS production in MG-63 cells. Furthermore, tomentosin-induced intracellular ROS decreased cell viability and induced apoptosis, cell cycle arrest, and DNA damage in MG-63 cells. Taken together, our results suggested that tomentosin exerted anti-carcinogenic effects in MG-63 cells by induction of intracellular ROS.
    Keywords:  osteosarcoma; reactive oxygen species (ROS); tomentosin
    DOI:  https://doi.org/10.3390/ijms20061508
  17. Cancer Sci. 2019 Mar 25.
    Wang Z, Shen J, Sun W, Zhang T, Zuo D, Wang H, Wang G, Xu J, Yin F, Mao M, Zhou Z, Hua Y, Cai Z.
      Osteosarcoma is the most common primary malignant bone tumor. Raddeanin A (RA) is an active oleanane-type triterpenoid saponin extracted from the traditional Chinese herb Anemone raddeana Regel that exerts antitumor activity against several cancer types. However, the effect of RA on osteosarcoma remains unclear. In the present study, we demonstrated that RA inhibited the proliferation and induced apoptosis of osteosarcoma cells in a dose- and time-dependent manner in vitro and in vivo. RA treatment resulted in excessive reactive oxygen species (ROS) generation and Jun amino-terminal kinase (JNK) and extracellular signal-regulated kinase (ERK1/2) activation. Apoptosis induction was evaluated by the activation of caspase-3, caspase-8, and caspase-9 and PARP cleavage. RA-induced cell death was significantly restored by the ROS scavenger glutathione (GSH), the pharmacological inhibitor of JNK SP600125, or specific JNK knockdown via shRNA. Additionally, signal transducer and activator of transcription 3 (STAT3) activation was suppressed by RA in human osteosarcoma, and this suppression was restored by GSH, SP600125, and JNK-shRNA. Further investigation showed that STAT3 phosphorylation was increased after JNK knockdown. In a tibial xenograft tumor model, RA induced osteosarcoma apoptosis and notably inhibited tumor growth. Taken together, our results reveal that RA suppresses proliferation and induces apoptosis by modulating the JNK/c-Jun and STAT3 signaling pathways in human osteosarcoma. Therefore, RA may be a promising candidate antitumor drug for osteosarcoma intervention. This article is protected by copyright. All rights reserved.
    Keywords:   JNK ; Osteosarcoma; Raddeanin A; SP600125; STAT3
    DOI:  https://doi.org/10.1111/cas.14008
  18. Cancer Cell Int. 2019 ;19 58
    Zhao Q, Wang W, Cui J.
      Background: Tumor necrosis factor-α (TNF-α) immunotherapy controls the progression of human cervical cancer. Here, we explored the detailed molecular mechanisms played by melatonin in human cervical cancer (HeLa cells) death in the presence of TNF-α injury, with a particular attention to the mitochondrial homeostasis.Methods: HeLa cells were incubated with TNFα and then cell death was determined via MTT assay, TUNEL staining, caspase ELISA assay and western blotting. Mitochondrial function was detected via analyzing mitochondrial membrane potential using JC-1 staining, mitochondrial oxidative stress using flow cytometry and mitochondrial apoptosis using western blotting.
    Results: Our data exhibited that treatment with HeLa cells using melatonin in the presence of TNF-α further triggered cancer cell cellular death. Molecular investigation demonstrated that melatonin enhanced the caspase-9 mitochondrion death, repressed mitochondrial potential, increased ROS production, augmented mPTP opening rate and elevated cyt-c expression in the nucleus. Moreover, melatonin application further suppressed mitochondrial ATP generation via reducing the expression of mitochondrial respiratory complex. Mechanistically, melatonin augmented the response of HeLa cells to TNF-α-mediated cancer death via repressing mitophagy. TNF-α treatment activated mitophagy via elevating Parkin expression and excessive mitophagy blocked mitochondrial apoptosis, ultimately alleviating the lethal action of TNF-α on HeLa cell. However, melatonin supplementation could prevent TNF-α-mediated mitophagy activation via inhibiting Parkin in a CaMKII-dependent manner. Interestingly, reactivation of CaMKII abolished the melatonin-mediated mitophagy arrest and HeLa cell death.
    Conclusions: Overall, our data highlight that melatonin enhances TNF-α-induced human cervical cancer HeLa cells mitochondrial apoptosis via inactivating the CaMKII/Parkin/mitophagy axis.
    Keywords:  CaMKII/Parkin pathways; HeLa cell; Melatonin; Mitochondria; TNF-α
    DOI:  https://doi.org/10.1186/s12935-019-0777-2
  19. Toxicology. 2019 Mar 21. pii: S0300-483X(18)30669-3. [Epub ahead of print]
    Ouyang L, Zhang W, Du G, Liu H, Xie J, Gu J, Zhang S, Zhou F, Shao L, Feng C, Fan G.
      Lead is widely distributed in the environment and has become a global public health issue. It is well known that lead exposure induces not only neurodevelopmental toxicity but also neurodegenerative diseases, with learning and memory impairment in the later stage. However, the molecular mechanisms remain elusive. The present study investigated the effects of early life and lifetime lead exposure on cognition and identified the molecular mechanisms involved in aged rats. The results herein demonstrated that the lead concentration in peripheral blood and brain tissues in aged rats was significantly increased in a lead dose-dependent manner. High-dose lead exposure caused cognitive functional impairment in aged rats, concomitant with a longer escape latency and a lower frequency of crossing the platform via Morris water maze testing compared to those in the control and low-dose lead exposure groups. Importantly, neuron functional defects were still observed even in early life lead exposure during the prenatal and weaning periods in aged rats. The neurotoxicity induced by lead exposure was morphologically evidenced by a recessed nuclear membrane, a swollen endoplasmic reticulum, and mitochondria in the neurons. Mechanistically, the exposure of aged rats to lead resulted in increasing free calcium concentration, reactive oxygen species, and apoptosis in the hippocampal neurons. Lead exposure increased RyR3 expression and decreased the levels of p-CaMKIIα/CaMKIIα and p-CREB/CREB in the hippocampus of aged rats. These findings indicated that early life lead exposure-induced cognition disorder was irreversible in aged rats. Lead-induced neurotoxicity might be related to the upregulation of RyR3 expression and high levels of intracellular free calcium with increasing lead concentration in injured neurons.
    Keywords:  RyR3; early life; lead; lifetime; neurotoxicity
    DOI:  https://doi.org/10.1016/j.tox.2019.03.005
  20. Bioorg Med Chem. 2019 Mar 22. pii: S0968-0896(18)30247-5. [Epub ahead of print]
    Machado FC, Adum de Matos RP, Primo FL, Tedesco AC, Rahal P, Calmon MF.
      Curcumin, a natural compound has several antineoplastic activities and is a promising natural photosensitizer used in photodynamic therapy. However, its low solubility in physiological medium limit the clinical use of curcumin. This study aimed to analyze the action of curcumin-nanoemulsion, a new and well-designed Drug Delivery System (DDS+) molecule, used as a photosensitizing agent in photodynamic therapy in an in vitro breast cancer model, MCF-7 cells. The empty nanoemulsion fulfils all necessary requirements to be an excellent DDS. Furthermore, the use of curcumin-nanoemulsion in photodynamic therapy resulted in a high phototoxic effect after activation at 440 nm, decreasing to <10% viable tumor cells after two irradiations and increasing the reactive oxygen species (ROS) production. The use of curcumin-nanoemulsion associated with photodynamic therapy resulted in an increase in the levels of caspase 3/7 activity for the studied MCF-7 cell model, indicating that this therapy triggers a cascade of events that lead to cell death, such as cellular apoptosis. In conclusion, curcumin-nanoemulsion proved to be efficient as a photosensitizing agent, had phototoxic effects, significantly decreased the proliferation of MCF-7 cells and stimulating the ROS production in combination with photodynamic therapy, so, this formulation has a great potential for use in treatment of breast cancer.
    Keywords:  Breast cancer; Curcumin; Nanotechnology; Photodynamic therapy
    DOI:  https://doi.org/10.1016/j.bmc.2019.03.044
  21. Biol Res. 2019 Mar 27. 52(1): 15
    Gwangwa MV, Joubert AM, Visagie MH.
      BACKGROUND: Tumourigenic cells modify metabolic pathways in order to facilitate increased proliferation and cell survival resulting in glucose- and glutamine addiction. Previous research indicated that glutamine deprivation resulted in potential differential activity targeting tumourigenic cells more prominently. This is ascribed to tumourigenic cells utilising increased glutamine quantities for enhanced glycolysis- and glutaminolysis. In this study, the effects exerted by glutamine deprivation on reactive oxygen species (ROS) production, mitochondrial membrane potential, cell proliferation and cell death in breast tumourigenic cell lines (MCF-7, MDA-MB-231, BT-20) and a non-tumourigenic breast cell line (MCF-10A) were investigated.RESULTS: Spectrophotometry demonstrated that glutamine deprivation resulted in decreased cell growth in a time-dependent manner. MCF-7 cell growth was decreased to 61% after 96 h of glutamine deprivation; MDA-MB-231 cell growth was decreased to 78% cell growth after 96 h of glutamine deprivation, MCF-10A cell growth was decreased 89% after 96 h of glutamine deprivation and BT-20 cell growth decreased to 86% after 24 h of glutamine deprivation and remained unchanged until 96 h of glutamine deprivation. Glutamine deprivation resulted in oxidative stress where superoxide levels were significantly elevated after 96 h in the MCF-7- and MDA-MB-231 cell lines. Time-dependent production of hydrogen peroxide was accompanied by aberrant mitochondrial membrane potential. The effects of ROS and mitochondrial membrane potential were more prominently observed in the MCF-7 cell line when compared to the MDA-MB-231-, MCF-10A- and BT-20 cell lines. Cell cycle progression revealed that glutamine deprivation resulted in a significant increase in the S-phase after 72 h of glutamine deprivation in the MCF-7 cell line. Apoptosis induction resulted in a decrease in viable cells in all cell lines following glutamine deprivation. In the MCF-7 cells, 87.61% of viable cells were present after 24 h of glutamine deprivation.
    CONCLUSION: This study demonstrates that glutamine deprivation resulted in decreased cell proliferation, time-dependent- and cell line-dependent ROS generation, aberrant mitochondrial membrane potential and disrupted cell cycle progression. In addition, the estrogen receptor positive MCF-7 cell line was more prominently affected. This study contributes to knowledge regarding the sensitivity of breast cancer cells and non-tumorigenic cells to glutamine deprivation.
    Keywords:  Apoptosis; Cell cycle progression; Glutamine deprivation; Mitochondrial membrane potential; ROS
    DOI:  https://doi.org/10.1186/s40659-019-0224-9
  22. Bioorg Med Chem. 2019 Mar 19. pii: S0968-0896(19)30161-0. [Epub ahead of print]
    Zeng A, Hua H, Liu L, Zhao J.
      Betulinic acid (BA) is a pentacyclic triterpenoids extracted from birch with a wide range of biological properties. Recent studies have shown that BA has significant cytotoxicity to various types of human cancer cells, and shows potential in cancer treatment. However, the efficacy of BA on human colorectal cancer tumor cells is still unclear. The purpose of our study was to evaluate the anti-cancer activity of BA in human colorectal cancer cells in vitro and in vivo to investigate the possible mechanism. In this experiment, we found that BA inhibited colorectal cancer cell lines in vitro with a time-dependent and dose-dependent manner. Moreover, BA could induce cell apoptosis by upregulating expression of Bax and cleaved caspase-3 and downregulating protein of Bcl-2. BA could increase the production of reactive oxygen species and reduce mitochondrial membrane potential of cancer cell, suggesting that BA induced cancer cells apoptosis by mitochondrial mediated pathways. Furthermore, BA significantly inhibited the migration and invasion of colorectal cancer cells, reduced the expression of matrix metalloproteinase (MMPs) and increased the expression of MMPs inhibitor (TIMP-2). In addition, the growth of tumor was significantly suppressed by intraperitoneal administration of 20 mg/kg/day of BA in a xenograft tumor mouse model of HCT-116. Histopathological and immunohistochemical analysis showed that MMP-2+ cells and Ki-67+ cells were reduced and cleaved caspase-3+ cells were increased in tumor tissues of mice after BA administration. The results showed that BA not only promoted the apoptosis of colorectal cancer cells, but also inhibited the metastasis of cancer cells. Our results suggest that BA can be a potential natural drug to inhibit the growth and metastasis of colorectal cancer.
    Keywords:  Apoptosis; Betulinic acid; Colorectal cancer cells; Invasion; Migration
    DOI:  https://doi.org/10.1016/j.bmc.2019.03.033
  23. Sci Rep. 2019 Mar 25. 9(1): 5039
    Lee SI, Jeong YJ, Yu AR, Kwak HJ, Cha JY, Kang I, Yeo EJ.
      Neuroblastoma is a solid malignant tumor of the sympathetic nervous system, which accounts for 8-10% of childhood cancers. Considering the overall high risk and poor prognosis associated with neuroblastoma, effective therapeutics should be developed to improve patient survival and quality of life. A recent study showed that a proteasome inhibitor, carfilzomib (CFZ), reduced cell viability of SK-N-BE(2)-M17 neuroblastoma cells. Therefore, we investigated the molecular mechanisms by which CFZ lower the cell viability of neuroblastoma cells. CFZ reduced cell viability via cell cycle arrest at G2/M and apoptosis, which involved caspase activation (caspases-8, 9, 4, and 3), endoplasmic reticulum stress, reactive oxygen species production, mitochondrial membrane potential loss, and autophagy in a dose- and time-dependent manner. The effect of CFZ was additive to that of cisplatin (Cis), a well-known chemotherapeutic drug, in terms of cell viability reduction, cell cycle arrest, and apoptosis. Importantly, the additive effect of CFZ was maintained in Cis-resistant neuroblastoma cells. These results suggest that CFZ can be used in combination therapy for patients with neuroblastoma to overcome the resistance and adverse side effects of Cis.
    DOI:  https://doi.org/10.1038/s41598-019-41527-0
  24. Sci Rep. 2019 Mar 26. 9(1): 5184
    Konari N, Nagaishi K, Kikuchi S, Fujimiya M.
      The underlying therapeutic mechanism of renal tubular epithelium repair of diabetic nephropathy (DN) by bone marrow-derived mesenchymal stem cells (BM-MSCs) has not been fully elucidated. Recently, mitochondria (Mt) transfer was reported as a novel action of BM-MSCs to rescue injured cells. We investigated Mt transfer from systemically administered BM-MSCs to renal proximal tubular epithelial cells (PTECs) in streptozotocin (STZ)-induced diabetic animals. BM-MSCs also transferred their Mt to impaired PTECs when co-cultured in vitro, which suppressed apoptosis of impaired PTECs. Additionally, BM-MSC-derived isolated Mt enhanced the expression of mitochondrial superoxide dismutase 2 and Bcl-2 expression and inhibited reactive oxygen species (ROS) production in vitro. Isolated Mt also inhibited nuclear translocation of PGC-1α and restored the expression of megalin and SGLT2 under high glucose condition (HG) in PTECs. Moreover, isolated Mt directly injected under the renal capsule of STZ rats improved the cellular morphology of STZ-PTECs, and the structure of the tubular basement membrane and brush border in vivo. This study is the first to show Mt transfer from systemically administered BM-MSCs to damaged PTECs in vivo, and the first to investigate mechanisms underlying the potential therapeutic effects of Mt transfer from BM-MSCs in DN.
    DOI:  https://doi.org/10.1038/s41598-019-40163-y
  25. Int J Biol Sci. 2019 ;15(4): 870-881
    Wang R, Xue X, Wang Y, Zhao H, Zhang Y, Wang H, Miao D.
      The polycomb repressor B lymphoma Mo-MLV insertion region 1 (BMI1) is a core composition of polycomb repressive complex 1 (PRC1) and contributes to diverse fundamental cellular processes including cell senescence, apoptosis and proliferation. To investigate the role and mechanism of BMI1 in maintaining normal female reproductive function, we compared the differences in reproductive phenotypes between Bmi1-deficient and wild-type female mice. The Bmi1-deficient female mice were then supplemented with N-acetylcysteine in their drinking water to explore whether antioxidant supplementation could improve reproductive dysfunction caused by BMI1 deficiency. The results revealed that Bmi1 deletion resulted in complete infertility in female mice, estrous cycle disorder, and follicular developmental disorders. The reactive oxygen species levels in the ovarian tissue were increased; the ability of antioxidant enzymes was downregulated; the expression levels of p19 and p53 proteins were significantly upregulated. We also found that oocytes derived from Bmi1-deficient mice could not develop into embryos by in vitro fertilization and in vitro culture of embryos. Furthermore, supplementation with the antioxidant NAC not only improved the reproductive defects caused by Bmi1 deletion, but also largely rescued the ability of Bmi1-deficient oocytes to develop into embryos in vitro. These results indicated that cells lacking Bmi1 resulted in female infertility by activating the p16/p19 signaling pathway, increasing oxidative stress and DNA damage, inhibiting granulosa cell proliferation, and inducing granulosa cell apoptosis. Thus, BMI1 may be a novel potential target for the clinical treatment of female infertility.
    Keywords:  BMI1; DNA damage; antioxidant; female infertility; oxidative stress
    DOI:  https://doi.org/10.7150/ijbs.30488
  26. Chem Biol Drug Des. 2019 Mar 28.
    Ma C, Wang Y, Dong F, Wang Z, Zhao Y, Shan Y, Gu W, Wang S.
      A series of novel isolongifoleno[7,8-d]thiazolo[3,2-a]pyrimidine derivatives (4a-4x) were synthesized from isolongifolanone according fragment-based design strategy, and their anticancer activity against human aortic smooth muscle cells (HASMC), human breast cancer (MCF-7) cells, human cervical cancer (HeLa) cells, and human liver cancer (HepG2) cells were investigated. Results of the anticancer activity illustrated that most of the compounds showed potent antitumor activity and compound 4i proved to be the most active derivative with IC50 values of 0.33 ± 0.24 (for MCF-7 cells), 0.52 ± 0.13 (for HeLa cells) and 3.09 ± 0.11 μM (for HepG2 cells), respectively. Moreover, we assessed the effects of 4i on cell apoptosis, cell cycle distribution, mitochondrial membrane potential and reactive oxygen species (ROS) generation. The results indicated that compound 4i altered mitochondrial membrane potential and produced ROS leading to cell apoptosis of MCF-7 cells in a dose-dependent manner, however, without affecting cell cycle progression. These findings suggested that 4i was an effective compound and provided a promising candidate for anti-cancer drugs. This article is protected by copyright. All rights reserved.
    Keywords:   ROS ; 2-a]pyrimidine derivatives; anticancer activity; apoptosis; isolongifolanone; mitochondria; thiazolo[3
    DOI:  https://doi.org/10.1111/cbdd.13522
  27. Oxid Med Cell Longev. 2019 ;2019 9706792
    Li X, Chen Y, Zhao J, Shi J, Wang M, Qiu S, Hu Y, Xu Y, Cui Y, Liu C, Liu C.
      Multiple signaling pathways including ERK, PI3K-Akt, and NF-κB, which are essential for onset and development of cancer, can be activated by intracellularly sustained high levels of H2O2 provided by elevated activity and expression of copper/zinc superoxide dismutase (SOD1) that catalyzes the dismutation of O2 •- into H2O2. Here, tests performed by the utilization of our designed specific SOD1 inhibitor LD100 on cancer and normal cells reveal that the signaling pathways and their crosstalk to support cancer cell growth are repressed, but the signaling pathways to promote cancer cell cycle arrest and apoptosis are stimulated by specific SOD1 inhibition-mediated ROS changes. These regulated pathways constitute an ROS signaling network that determines the fate of cancer cells. This ROS signaling network is also regulated in SOD1 knockdown cells. These findings might facilitate disclosure of action mechanisms by copper-chelating anticancer agents and design of SOD1-targeting and ROS signaling pathway-interfering anticancer small molecules.
    DOI:  https://doi.org/10.1155/2019/9706792
  28. Cell Biochem Funct. 2019 Mar 25.
    Wang Y, Xue J, Li Y, Zhou X, Qiao S, Han D.
      Telmisartan, an angiotensin II receptor blocker, has been widely used for hypertension. It has also been reported to improve insulin sensitivity in animal models of obesity and diabetic patients by targeting to the peroxisome proliferator-activated receptor (PPAR)-γ. High glucose/high lipid (HG/HL)-induced apoptosis of pancreatic β-cells impairs its function of insulin secretion and is generally believed to be the key factor in the development of diabetes. In this study, we investigated whether telmisartan exerted a protective effect against HG/HL-induced apoptosis and insulin secretion in vitro as well as in vivo; 10-μM telmisartan treatment significantly reduced HG (25 mM) or/and HL (0.4 mM palmitic acid) induced-cell apoptosis and greatly improved insulin secretion in INS-1 pancreatic β-cells, which is consistent in an obesity rat model induced by HG/HL diets. Furthermore, telmisartan treatment markedly reduced the protein level of GRP78, CHOP, and caspase 12, while increasing anti-apoptotic Bcl-2 protein expression. Moreover, telmisartan treatment significantly reduced intracellular ROS levels. Mechanistically, we demonstrated that PPARγ signaling pathway may be involved in the telmisartan protective effects, which were blocked by a PPARγ blocker, GW9662. In conclusion, the protective effect of telmisartan was mediated by an anti-ER stress-induced apoptotic and anti-oxidative pathway. SIGNIFICANCE OF THIS STUDY: Type 2 diabetes mellitus (T2DM) is a chronic metabolic disorder worldwide pathologically characterized by hyperglycemia and insulin resistance. Long-term high glucose in the blood has been proposed to induce pancreatic β-cell loss and is generally believed to be the key factor in the development of diabetes. In the present study, we demonstrated that telmisartan, a common drug used for hypertension treatment, has a protective effect against high glucose/high lipid-induced cell apoptosis and greatly improves the insulin secretion function by inhibiting the oxidative stress and ER stress. Furthermore, this protective effect of telmisartan is mediated by the PPAR-γ signal pathway, which may provide a potential strategy against T2DM.
    Keywords:  ER stress; apoptosis; high glucose/high lipid; insulin secretion; oxidative stress; telmisartan
    DOI:  https://doi.org/10.1002/cbf.3383
  29. Redox Rep. 2019 Dec;24(1): 17-26
    Ríos-Ocampo WA, Daemen T, Buist-Homan M, Faber KN, Navas MC, Moshage H.
      OBJECTIVES: The occurrence of oxidative stress and endoplasmic reticulum (ER) stress in hepatitis C virus (HCV) infection has been demonstrated and play an important role in liver injury. During viral infection, hepatocytes must handle not only the replication of the virus, but also inflammatory signals generating oxidative stress and damage. Although several mechanisms exist to overcome cellular stress, little attention has been given to the adaptive response of hepatocytes during exposure to multiple noxious triggers.METHODS: In the present study, Huh-7 cells and hepatocytes expressing HCV Core or NS3/4A proteins, both inducers of oxidative and ER stress, were additionally challenged with the superoxide anion generator menadione to mimic external oxidative stress. The production of reactive oxygen species (ROS) as well as the response to oxidative stress and ER stress were investigated.
    RESULTS: We demonstrate that hepatocytes diminish oxidative stress through a reduction in ROS production, ER-stress markers (HSPA5 [GRP78], sXBP1) and apoptosis (caspase-3 activity) despite external oxidative stress. Interestingly, the level of the autophagy substrate protein p62 was downregulated together with HCV Core degradation, suggesting that hepatocytes can overcome excess oxidative stress through autophagic degradation of one of the stressors, thereby increasing cell survival. Duscussion: In conclusion, hepatocytes exposed to direct and indirect oxidative stress inducers are able to cope with cellular stress associated with viral hepatitis and thus promote cell survival.
    Keywords:  Core; ER stress; Hepatitis C virus; Transient expression; apoptosis; cellular stress; nS3/4A; oxidative stress; unfolded protein response
    DOI:  https://doi.org/10.1080/13510002.2019.1596431
  30. J Photochem Photobiol B. 2019 Mar 21. pii: S1011-1344(18)31466-0. [Epub ahead of print]194 46-55
    Xie J, Wang S, Li Z, Ao C, Wang J, Wang L, Peng X, Zeng K.
      Human papillomavirus (HPV) infection is linked to several diseases, the most prominent of which are cervical cancer and genital condyloma acuminatum. Previous studies have suggested an effective role for 5-aminolevulinic acid photodynamic therapy (ALA-PDT) against various cancers by the induction of autophagy and apoptosis. However, few reports have focused on the effectiveness of ALA-PDT on HPV related disorders. To identify the role of ALA-PDT in the context of HPV infection, we initially investigated 111 patients suffering from genital condyloma acuminatum. HPV viral load detected before and after ALA-PDT treatment was compared during this procedure; a significant difference was noted. HeLa (HPV18) cells were exposed to ALA-PDT in vitro to further explore the underlying mechanisms. Western blot analysis showed that ALA-PDT induces LC3II and p62 expression, along with the up regulation of caspase-3 and cleaved caspase-3. Our study also demonstrated that ALA-PDT treatment inhibits the proliferation of HeLa cells in a dose dependent manner and effectively reduces HPV viral load via autophagy and apoptosis by regulating the Ras/Raf/MEK/ERK and PI3K/AKT/mTOR pathways. Hydroxychloroquine (HCQ), although it inhibited autophagy degradation, functioned to activate reactive oxygen species (ROS) levels of ALA-PDT to enhance the observed effect. These findings suggest strategies for the improvement of PDT efficacy in patients.
    Keywords:  ALA-PDT; Apoptosis; Autophagy; HPV; HeLa
    DOI:  https://doi.org/10.1016/j.jphotobiol.2019.03.012
  31. Exp Dermatol. 2019 Mar 29.
    Xu M, Hua Y, Qi Y, Meng G, Yang S.
      Hydrogen sulfide (H2 S) is an important gasotransmitter with several physiological functions. However, the roles and the detailed mechanisms of H2 S on skin wound healing are not known well. In the present study, 129S1/SvImJ mice were intraperitoneally injected with NaHS (50 μmol/kg/d) for 2 weeks. Then a round wound of 6 mm diameter with depth into the dermis was made. The skin wound area, blood perfusion, superoxide production, malondialdehyde (MDA) levels, total antioxidant capacity (T-AOC), expression of vascular endothelial growth factor (VEGF), dynamin-related protein 1 (DRP1) and optic atrophy 1 (OPA1) were measured. After NaHS (50 μmol/L) pre-administration for 4 h, cell migration rate, DRP1, OPA1 and α-smooth muscle actin (α-SMA) expression, superoxide production and mitochondrial membrane potential in primary skin fibroblasts were measured. Tube formation in human umbilical vein endothelial cells (HUVECs) and cell migration in human keratinocytes were also measured. The results showed that NaHS pre-treatment significantly accelerated wound healing and improved blood flow in the wound after operation. NaHS increased VEGF expression in the wound and promoted tube formation in HUVECs. Meanwhile, NaHS attenuated reactive oxygen species (ROS) production, suppressed MDA level but restored T-AOC in the wound. NaHS also promoted skin fibroblasts migration and α-SMA expression after scratch. Moreover, NaHS alleviated ROS, increased mitochondrial membrane potential, decreased DRP1 but enhanced OPA1 expression in skin fibroblasts after scratch. NaHS also accelerated human keratinocytes migration after scratch. Taken together, exogenous H2 S supplementary accelerated the skin wound healing, which might be related to oxidative stress inhibition and VEGF enhancement. This article is protected by copyright. All rights reserved.
    Keywords:  hydrogen sulfide; reactive oxygen species; skin fibroblasts; vascular endothelial growth factor; wound healing
    DOI:  https://doi.org/10.1111/exd.13930
  32. J Cell Physiol. 2019 Mar 25.
    Mortezaee K, Najafi M, Farhood B, Ahmadi A, Shabeeb D, Musa AE.
      Radiotherapy and chemotherapy are two famous modalities in tumor-targeted therapy that lead to systemic and local toxicities for normal tissues. Moreover, several studies have confirmed that exposure of the tumor to radiation or chemotherapy drugs stimulate some signaling pathways in the tumor microenvironment (TME), leading to resistance of cancer cells to apoptosis, as well as promoting angiogenesis and tumor growth. Nuclear factor kappa B (NF-κB) plays a central role in the regulation of inflammatory responses in both normal tissues and tumors via the release of several cytokines, regulation of prostaglandins, reduction/oxidation (redox) reactions, angiogenesis, and cell death. Upregulation of NF-κB in normal tissues causes an appearance of inflammatory reactions and oxidative stress, whereas it regulates angiogenesis and suppresses apoptosis, leading to resistance to subsequent doses of radiation or chemotherapy. Selective inhibition of NF-κB in experimental studies has shown promising results for tumor sensitization via apoptosis induction, inhibition of angiogenesis, and increasing delay of tumor growth. The use of some agents for NF-κB inhibition has been shown to alleviate radiation/chemotherapy toxicities in normal cells/ tissues. In this current review, we explained the pivotal role of NF-κB in both normal tissue toxicity and tumor resistance. We also discussed the promising strategies for overcoming these problems with regard to chemotherapy and radiotherapy.
    Keywords:  chemotherapy; normal tissue toxicity; nuclear factor kappa B (NF-κB); radiotherapy; tumor resistance
    DOI:  https://doi.org/10.1002/jcp.28504
  33. J Diabetes Res. 2019 ;2019 8520856
    Zhao B, Yuan Q, Hou JB, Xia ZY, Zhan LY, Li M, Jiang M, Gao WW, Liu L.
      Background: A substantial increase in histone deacetylase 3 (HDAC3) expression is implicated in the pathological process of diabetes and stroke. However, it is unclear whether HDAC3 plays an important role in diabetes complicated with stroke. We aimed to explore the role and the potential mechanisms of HDAC3 in cerebral ischemia/reperfusion (I/R) injury in diabetic state.Methods: Diabetic mice were subjected to 1 h ischemia, followed by 24 h reperfusion. PC12 cells were exposed to high glucose for 24 h, followed by 3 h of hypoxia and 6 h of reoxygenation (H/R). Diabetic mice received RGFP966 (the specific HDAC3 inhibitor) or vehicle 30 minutes before the middle cerebral artery occlusion (MCAO), and high glucose-incubated PC12 cells were pretreated with RGFP966 or vehicle 6 h before H/R.
    Results: HDAC3 inhibition reduced the cerebral infarct volume, ameliorated pathological changes, improved the cell viability and cytotoxicity, alleviated apoptosis, attenuated oxidative stress, and enhanced autophagy in cerebral I/R injury model in diabetic state in vivo and in vitro. Furthermore, we found that the expression of HDAC3 was remarkably amplified, and the Bmal1 expression was notably decreased in diabetic mice with cerebral I/R, whereas this phenomenon was obviously reversed by RGFP966 pretreatment.
    Conclusions: These results suggested that the HDAC3 was involved in the pathological process of the complex disease of diabetic stroke. Suppression of HDAC3 exerted protective effects against cerebral I/R injury in diabetic state in vivo and in vitro via the modulation of oxidative stress, apoptosis, and autophagy, which might be mediated by the upregulation of Bmal1.
    DOI:  https://doi.org/10.1155/2019/8520856
  34. Toxicol In Vitro. 2019 Mar 21. pii: S0887-2333(18)30555-1. [Epub ahead of print]58 118-125
    Fraser DL, Stander BA, Steenkamp V.
      As knowledge regarding mechanisms of pentachlorophenol (PCP) toxicity in neuronal cell lines is limited, the aim of the study was to evaluate the effects of PCP and its active metabolites, tetrachloro-1,4-benzoquinone (TCBQ) and tetrachlorohydroquinone (TCHQ) in human neuroblastoma SH-SY5Y cells. All compounds induced cytotoxic effects in time- and dose-dependent manners, and resulted in differential modes of cell death. Reduced mitochondrial membrane potential (ΔᴪM) and oxidative damage lead to apoptosis and necrosis following TCBQ and PCP exposure, respectively. Time-dependent investigations revealed transient ΔᴪM recovery in TCHQ exposed cells, and redox stress. Sufficient ΔᴪM recovery allowed apoptosis completion in TCHQ exposed cells, whereas overwhelming metabolic and oxidative stress saw a conversion from apoptotic to necrotic-like cell death. The onset of mitochondrial dysfunction preceded that of redox damage for all compounds, indicating that oxidative damage is secondary to ΔᴪM insult. Cytotoxic events were further linked to cell cycle. S phase and G2/M blocks were observed after 12 h exposure to TCBQ and TCHQ, respectively, while a G1 block occurred after 24 h exposure to PCP. This study provides new insight regarding time-dependant toxic effects of PCP and its metabolites in human neuronal cells.
    Keywords:  Cell death; Mitochondrial membrane potential; Oxidative stress; Pentachlorophenol; Tetrachloro-1,4-benzoquinone; Tetrachlorohydroquinone
    DOI:  https://doi.org/10.1016/j.tiv.2019.03.024
  35. Oxid Med Cell Longev. 2019 ;2019 3231424
    Yang X, He T, Han S, Zhang X, Sun Y, Xing Y, Shang H.
      Oxidative stress has been closely related with coronary artery disease. In coronary heart disease (CHD), an excess of reactive oxygen species (ROS) production generates endothelial cell and smooth muscle functional disorders, leading to a disequilibrium between the antioxidant capacity and prooxidants. ROS also leads to inflammatory signal activation and mitochondria-mediated apoptosis, which can promote and increase the occurrence and development of CHD. There are several kinds of antioxidative and small molecular systems of antioxidants, such as β-carotene, ascorbic acid, α-tocopherol, and reduced glutathione (GSH). Studies have shown that antioxidant treatment was effective and decreased the risk of CHD, but the effect of the treatment varies greatly. Traditional Chinese medicine (TCM) has been utilized for thousands of years in China and is becoming increasingly popular all over the world, especially for the treatments of cardiovascular diseases. This review will concentrate on the evidence of the action mechanism of TCM in preventing CHD by modulating oxidative stress-related signaling pathways.
    DOI:  https://doi.org/10.1155/2019/3231424
  36. Mediators Inflamm. 2019 ;2019 6453296
    Wang QL, Yang L, Peng Y, Gao M, Yang MS, Xing W, Xiao XZ.
      Objectives: To investigate the protective effect of ginsenoside Rg1 on relieving sepsis-induced lung inflammation and injury in vivo and in vitro.Methods: Cultured human pulmonary epithelial cell line A549 was challenged with LPS to induce cell injury, and CLP mouse model was generated to mimic clinical condition of systemic sepsis. Rg1 was applied to cells or animals at indicated dosage. Apoptosis of cultured cells was quantified by flow cytometry, along with ELISA for inflammatory cytokines in supernatant. For septic mice, lung tissue pathology was examined, plus ELISA assay for serum cytokines. Western blotting was used to examine the activation of inflammatory pathways and ER stress marker proteins in both cells and mouse lung tissues. Reactive oxygen species (ROS) level was quantified by DCFDA kit.
    Results: Ginsenoside Rg1 treatment remarkably suppressed apoptosis rate of LPS-induced A549 cells, relieved mouse lung tissue damage, and elevated survival rate. Rg1 treatment also rescued cells from LPS-induced intracellular ROS. In both A549 cells and mouse lung tissues, further study showed that Rg1 perfusion significantly suppressed the secretion of inflammatory cytokines including tumor necrosis factor- (TNF-) alpha and interleukin- (IL-) 6 and relieved cells from ER stress as supported by decreased expression of marker proteins via upregulating sirtuin 1 (SIRT1).
    Conclusion: Our results showed that ginsenoside Rg1 treatment effectively relieved sepsis-induced lung injury in vitro and in vivo, mainly via upregulating SIRT1 to relieve ER stress and inflammation. These findings provide new insights for unrevealing potential candidate for severe sepsis accompanied with lung injury.
    DOI:  https://doi.org/10.1155/2019/6453296
  37. J Mol Neurosci. 2019 Mar 30.
    Zhou H, Li S, Wang G.
      Sevoflurane is a widely used anesthetic. A series of recent studies have shown that exposure to sevoflurane at an early stage is a risk factor for the development of learning and memory dysfunction. Euxanthone is a xanthone derivative obtained from Polygala caudata. This study was designed to investigate whether euxanthone can confer neuroprotective activities against sevoflurane-induced neurotoxicity and to determine the associated molecular mechanisms. Neonatal Sprague-Dawley (male) rats were exposed to sevoflurane with or without euxanthone treatment. The behavioral data of rats were collected at P41 (the beginning of the adult stage). The hippocampal tissue was obtained following exposure to sevoflurane. The reactive oxygen species (ROS) level in the hippocampal tissue was determined by a commercial kit. The expression of apoptotic markers and inflammatory cytokines was determined by western blot. The mRNA and protein expression of Nrf2 were determined by qRT-PCR and western blot, respectively. The rat in vitro model of neurotoxicity was established using isolated hippocampal neurons. Nrf2 expression was repressed by transfection of siRNA. The cell viability was assessed by the CCK-8 assay. The flow cytometry was performed to measure apoptotic cell death. Our data showed that euxanthone treatment at the neonatal stage protected against sevoflurane-induced neurotoxicity in adult rats. At the molecular level, our findings revealed that the neuroprotective activities of euxanthone were associated with decreased sevoflurane-induced apoptosis cell death and neuroinflammation. More importantly, our results provide the experimental evidence that euxanthone confers neuroprotection by upregulating Nrf2 expression. Euxanthone has a therapeutic potential for clinical prevention of sevoflurane-induced neurotoxicity.
    Keywords:  Apoptosis; Euxanthone; Neuroinflammation; Neurotoxicity; Nrf2; Sevoflurane
    DOI:  https://doi.org/10.1007/s12031-019-01303-1
  38. Anal Chim Acta. 2019 Jul 11. pii: S0003-2670(19)30165-5. [Epub ahead of print]1061 142-151
    Zhang K, Lv Y, Meng J, Wang J, Peng A, Wang X, Tian Z.
      A single-chromophore-based photoactive agent (MB-DOPA) capable of rapid sensing of nanomolar hypochlorous acid (HOCl) and in-situ generating photocytotoxicity to cancer cells was developed using dopamine moiety as the recognition unit and methylene blue (MB) moiety as the fluorescence signaling unit. Specifically, HOCl triggered conversion of the nonfluorescent MB-DOPA to MB enabling far-red fluorescence emission (λmax ∼ 683 nm) and additional ability to photogenerate 1O2 species. Owing to the catechol nature of dopamine characterized with strong electron-donating property, MB-DOPA underwent HOCl-mediated conversion with response time of ∼20 s and a strong fluorescence OFF-to-ON contrast by a factor of more than 3000. The preliminary bioimaging results confirmed the intracellular HOCl sensing ability of MB-DOPA and the in-situ photodynamic therapy (PDT) effectiveness for inducing massive apoptosis of cancer cells. The figure of merits of MB-DOPA, including ability for sensing of nanomolar HOCl with high specificity, rapid response, practicality for intracellular fluorescence imaging, and the in-situ generation of 1O2 for killing tumor cells, is expected to enable diagnosis of early-stage oncogenesis based on the highly specific detection of abnormal HOCl levels in the transformed cells and the simultaneous treatment in biomedical applications.
    Keywords:  Fluorescence; Hypochlorous acid; Photodynamic therapy; Reactive oxygen species; Tumor
    DOI:  https://doi.org/10.1016/j.aca.2019.02.005
  39. J Biol Inorg Chem. 2019 Mar 29.
    Hajebi S, Tabrizi MH, Moghaddam MN, Shahraki F, Yadamani S.
      Based on recent researches, bio synthesized silver nanoparticles (Ag-NPs) seem to have the potential in declining angiogenesis and oxidative stress. In the current study, rapeseed flower pollen (RFP) water extract was triggered to synthesize RFP-silver nanoparticles (RFP/Ag-NPs). Moreover, antioxidant, antiangiogenesis and cytotoxicity of the RFP/Ag-NPs against MDA-MB-231, MCF7 and carcinoma cell lines and normal human skin fibroblast HDF were compared. Results indicated that RFP/Ag-NPs have a peak at 430 nm, spherical shape and an average size of 24 nm. According to the results of FTIR, rapeseed pollen capped Ag-NPs. RFP/Ag-NPs have cytotoxicity on MDA-MB-231 and MCF7 cells and decrease cancerous cell viability (IC50 = 3 µg/ml and 2 µg/ml, respectively) in a dose- and time-dependent manner. The morphological data showed that the RFP/Ag-NPs increase the percentage of apoptotic cells compared to the control group and normal cells (human skin fibroblast cells). The apoptotic morphological change was also confirmed with a flow cytometric analysis. RFP/ Ag-NPs' antioxidant activity was evaluated by measuring their ability to scavenge ABTS and DPPH free radicals. The IC50 values were determined at 800 and 830 μg/ml for ABTS and DPPH tests, respectively. According to the results, green-synthesized RFP/Ag-NPs as a safe efficient apoptosis inducer and strong antioxidant compound have the potential to suppress breast cancer carcinogenesis by VEGF down-regulatiion and thus sensitizing them against apoptosis. However, further researches are required to clarify RFP/Ag-NPs' cell specificity and therapeutic doses in in vivo conditions.
    Keywords:  Antioxidant; Apoptosis-inducer; Breast cancerous cell line (MDA-MB-231); Green synthesized silver nanoparticles; Rapeseed flower pollen; VEGF down-regulating
    DOI:  https://doi.org/10.1007/s00775-019-01655-4
  40. Cell Death Discov. 2019 ;5 79
    Liu Z, Xu Y, Wan Y, Gao J, Chu Y, Li J.
      Exosomes from bone marrow stem cells or cardiac progenitor cells can reduce apoptosis in myocardial cells after ischemia and reperfusion injury. However, there is little known about the effects of exosomes from adipose-derived stem cells (ADSCs), which are more abundant and have a lower risk of side effects. The aim of this study was to characterize exosomes from ADSCs and evaluate their cardioprotective actions against ischemia reperfusion injury. The exosomes were isolated from ADSCs and analyzed by protein marker expression, transmission electron microscopy, and nanoparticle tracking analysis. The ADSC-exosomes were then used for ex vivo investigation of the cardioprotective effects on cardiomyocytes after exposure to oxidative stress. Exosomes from ADSCs exhibited a diameter of 150 nm and expressed the marker proteins, CD9 and CD29. ADSC-exosomes had no effect on proliferation of untreated cardiomyocytes. In contrast, ADSC-derived exosomes reduced apoptosis in myocardial cells subjected to oxidative stress. This study confirms that exosomes originating from ADSCs can protect cardiomyocytes from oxidative stress.
    DOI:  https://doi.org/10.1038/s41420-019-0159-5
  41. Trends Plant Sci. 2019 Mar 22. pii: S1360-1385(19)30070-6. [Epub ahead of print]
    Kazan K, Kalaipandian S.
      Cell death is one of the most fundamental biological processes operating in multicellular organisms. Recent research highlighted here [Distéfano et al. (J. Cell Biol. 2017:216;463-476) and Dangol et al. (Plant Cell 2019:31;189-209)] revealed an iron- and ROS-dependent cell death phenomenon called ferroptosis in plants. Features distinguishing ferroptosis from other cell death events and how ferroptosis can be exploited to improve plant performance are discussed.
    Keywords:  apoptosis; autophagy; cell death; hypersensitive response; iron; reactive oxygen species
    DOI:  https://doi.org/10.1016/j.tplants.2019.03.005
  42. Biochem Cell Biol. 2019 Mar 29.
    Aminzadeh A, Salarinejad A.
      Lead is a major environmental pollutant, which causes serious adverse effects on biological systems and cells. In this study, we examined the effect of citicoline on lead-induced apoptosis in PC12 cells. PC12 cells were pre-treated with citicoline and then exposed to lead for 48 h. The effect of citicoline on cell survival was examined by MTT assay. In addition, lipid peroxidation (LPO), total thiol groups, total antioxidant power (TAP), catalase (CAT), superoxide dismutase (SOD), and reduced glutathione (GSH) were evaluated. The levels of Bax, Bcl-2, and caspase-3 were also measured by western blot analysis. Citicoline could significantly increase the cell viability of PC12 cells exposed to lead. Treatment of PC12 cells with lead increased LPO, while citicoline effectively decreased LPO. The total thiol groups, and TAP, CAT, SOD, and GSH were significantly increased in citicoline-treated PC12 cells compared to the lead group. Citicoline pretreatment significantly reduced Bax expression and increased the level of Bcl-2 expression. Citicoline also reduced caspase-3 activation in PC12 cells compared to the lead group. Our findings revealed that citicoline exerts a neuroprotective effect against lead-induced injury in PC12 cells through mitigation of oxidative stress and at least in part, through suppression of mitochondrial-mediated apoptotic pathway.
    DOI:  https://doi.org/10.1139/bcb-2018-0218
  43. Malays J Med Sci. 2018 Mar;25(2): 72-81
    Alidadi H, Khorsandi L, Shirani M.
      Background: Recent studies have demonstrated that many nanoparticles have an adverse or toxic effect on the kidney.Objective: To investigate the nephroprotective effect of quercetin (QT) against renal injury induced by titanium dioxide nanoparticles (NTiO2) in rats.
    Methods: NTiO2-intoxicated rats received 50 mg/kg of NTiO2 for seven days. The QT + NTiO2 group was pretreated with QT for seven days before being administered NTiO2. Uric acid, creatinine, and blood urea nitrogen were considered to be biomarkers of nephrotoxicity. Catalase (CAT) and superoxide dismutase (SOD) activities and renal levels of malondialdehyde (MDA) were measured to assess the oxidative stress caused by NTiO2.
    Results: NTiO2 significantly increased the plasma level of the biomarkers. It also significantly decreased the activities of CAT (P = 0.008) and SOD (P = 0.004), and significantly increased the MDA levels (P = 0.007). NTiO2 caused proximal tubule damage, the accumulation of red blood cells, the infiltration of inflammatory cells, and reduced the glomerular diameters, as well as induced apoptosis in the proximal tubules. Pre-treatment with QT attenuated the histological changes, normalised the plasma biomarkers, suppressed oxidative stress, ameliorated the activities of CAT (P = 0.007) and SOD (P = 0.006), and reduced apoptosis (P < 0.001).
    Conclusion: QT was found to have a potent protective effect against nephrotoxicity induced by NTiO2 in rats. It also reduced apoptosis caused by NTiO2.
    Keywords:  anti-oxidants; nanoparticles; nephrotoxicity; oxidative stress; quercetin; rats
    DOI:  https://doi.org/10.21315/mjms2018.25.2.8
  44. Am J Physiol Heart Circ Physiol. 2019 Mar 29.
    Choi H, Stark RJ, Raja BS, Dikalova A, Lamb FS.
      Tumor necrosis factor-α (TNFα) is pro-inflammatory cytokine that is closely linked to the development of cardiovascular disease. TNFα activates NADPH oxidase 1 (Nox1), and reactive oxygen species (ROS) including superoxide (O2•-) production extracellularly are required for subsequent signaling in vascular smooth muscle cells (VSMC). Apoptosis signal-regulating kinase 1 (ASK1) is a MAPKKK that is activated by oxidation of associated thioredoxin. The role of ASK1 in Nox1-mediated signaling by TNFα is poorly defined. We hypothesized that ASK1 is required for TNFα receptor endocytosis and subsequent inflammatory TNFα signaling. We employed a knockdown strategy to explore the role of ASK1 in TNFα signaling in VSMCs. Small interfering RNA (siRNA) targeting ASK1 had no effect on TNFα-induced extracellular O2•- production. However, siASK1 inhibited receptor endocytosis, as well as phosphorylation of two endocytosis-related proteins, dynamin1 and caveolin1. Intracellular O2•- production was subsequently reduced, as were other inflammatory signaling steps including; NF-κB activation, IL-6 production, iNOS and VCAM expression and VSMC proliferation. Prolonged exposure to TNFα (24 hours) increased TNFR subtype 1 and 2 expression and these effects were also attenuated by siASK1. ASK1 co-immunoprecipitated with both Nox1 and the LRRC8A anion channel, two essential components of the TNFR1 signaling complex. Activation of ASK1 by autophosphorylation at Thr845 occurs following thioredoxin dissociation and this requires the presence of Nox1. Thus, Nox1 is part of the multi-protein ASK1 signaling complex. In response to TNFα, ASK1 is activated by Nox1-derived oxidants and this plays a critical role in translating these ROS into a physiologic response in VSMCs.
    Keywords:  Apoptosis signal-regulating kinase 1; NADPH oxidase 1; Tumor Necrosis Factor-α; endocytosis; vascular smooth muscle cells
    DOI:  https://doi.org/10.1152/ajpheart.00741.2018
  45. J Ethnopharmacol. 2019 Mar 25. pii: S0378-8741(18)34973-0. [Epub ahead of print] 111836
    Gao S, Huang Q, Tang X, Qu X, Yu Y, Zhao Y, Tian L, Wu P, Gong H, Xu Y, Xu J.
      ETHNOPHARMACOLOGICAL RELEVANCE: ZhiShiXiaoPi Tang (ZSXPT) is a Chinese traditional medicine formula that contains 10 Chinese traditional medicine substances. It has been widely used to treat patients with functional dyspepsia (FD). However, the protective effect of ZSXPT and its molecular mechanisms in FD still remain elusive.AIM OF THE STUDY: To investigate the protective effect of ZSXPT on autophagy induced by Corticosterone (Cort) in PC12 cells which have typical neuron characteristics and have been widely used as a model system for depression studies and FD rats, and explore its underlying mechanisms.
    MATERIALS AND METHODS: In this study, high-performance liquid chromatography fingerprint analysis was performed to characterize the chemical composition of ZSXPT. Depression-induced autophagy, ROS generation, and changes in mitochondrial membrane potential (MMP) were investigated in Cort-induced PC12 cells and in FD rats to evaluate the protective effects of ZSXPT.
    RESULTS: Our results show that ZSXPT treatment protects neurons against Cort-induced damage and apoptosis by increasing cell viability and reducing the release of lactate dehydrogenase. ZSXPT decreased Cort-induced ROS generation, increased MMP, and accelerated autophagy through the blockade of the mammalian target of rapamycin (mTOR) pathway. Moreover, we observed similar findings when we studied ZSXPT in a rat model of FD.
    CONCLUSIONS: Our in vitro and in vivo results indicate that the neuroprotective effect of ZSXPT against autophagy-induced damage and apoptosis occurs mainly by blocking the mTOR pathway in Cort-induced PC12 cells and in FD rats. Taken together, these data provide reliable experimental evidence and explain the molecular mechanism by which ZSXPT ameliorates FD.
    Keywords:  Apoptosis; Autophagy; Depression; Functional dyspepsia; ZhiShiXiaoPi tang; mTOR pathways
    DOI:  https://doi.org/10.1016/j.jep.2019.111836
  46. J Steroid Biochem Mol Biol. 2019 Mar 21. pii: S0960-0760(18)30751-9. [Epub ahead of print]
    Tong T, Liu Z, Zhang H, Sun J, Zhang D, Wang F, Miao D, Shen Y.
      Accumulating evidence shows that genetic polymorphism of the vitamin D receptor (VDR) gene is associated with intervertebral disc degeneration (IDD), implying that VDR may be involved in the pathogenesis of IDD. However, the exact relationship between VDR and IDD remains unknown. The aim of this study was to investigate the age-dependent expression of VDR in rat intervertebral discs and to determine the effect of VDR on oxidative stress-induced cell apoptosis of the annulus fibrosus (AF) and the underlying mechanism. Sprague-Dawley rats were subjected to magnetic resonance imaging (MRI) and CT scans at young (2-3 months), adult (6-7 months), and old (14-15 months) ages. The images revealed age-related degeneration of the lumbar intervertebral discs and endplates. Immunohistochemistry demonstrated positive expression of VDR in the AF. The expression level of VDR in aged rats was significantly reduced compared with that in the young and adult animals and exhibited a negative correlation to IDD severity. Western blot analysis further demonstrated that the amount of VDR protein was significantly decreased in severe degenerative discs. AF cells were also isolated from young rat lumbar discs and subjected to different concentrations of hydrogen peroxide (H2O2) for various amounts of time. The results revealed that H2O2 inhibited the viability of AF cells and induced mitochondrial pathway apoptosis. However, pretreatment of AF cells with 10-7 and 10-8 M 1,25-dihydroxyvitamin D3 [1,25(OH)2D3] effectively increased cell viability, increased mitochondrial membrane potential, decreased the level of reactive oxygen species, increased mitochondrial ATP content, reserved the activity of key enzymes in the oxidative respiratory chain, and thus protected the mitochondria from H2O2-induced damage. Whereas, siRNA knock-down of VDR abolished the protective effects of 1,25(OH)2D3. Moreover, 1,25(OH)2D3 inhibited H2O2-induced autophagy of AF cells through inhibition of the mTOR/p70S6K signal pathway. Our study demonstrated that decreased expression of VDR may play a role in age-related intervertebral disc degeneration in rats and that activation of VDR ameliorates oxidative stress-induced apoptosis in AF cells by preserving mitochondrial functions.
    Keywords:  annulus fibrosus; apoptosis; intervertebral disc degeneration; mTOR/p70S6K signal pathway; oxidative stress; vitamin D receptor
    DOI:  https://doi.org/10.1016/j.jsbmb.2019.03.013
  47. Life Sci. 2019 Mar 20. pii: S0024-3205(19)30210-3. [Epub ahead of print]
    Deng T, Wang Y, Wang C, Yan H.
      Endoplasmic reticulum (ER) stress and subsequent apoptosis play a vital role in myocardial ischemia reperfusion (IR) injury. Fatty acid binding protein 4 (FABP4) may induce ER stress. The aim of this study was to investigate the mechanism and effect of FABP4 on IR injury in vitro. Rat H9c2 cells were exposed to hypoxia reoxygenation (HR) to create an IR model in vitro. FABP4 was overexpressed in HR-injured H9c2 cells. Transfection with FABP4 siRNA increased cell viability and decreased LDH upon HR stimulation. FABP4 cessation also suppressed apoptotic cells and caspase-3 activity after HR. Downregulation of FABP4 significantly inhibited ER stress by decreasing the protein expression of p-PERK, GRP78, and ATF6. FABP4 silencing also restrained the ER stress-mediated apoptotic pathway, as indicated by decreased pro-apoptotic proteins p-JNK, CHOP, Bax, and caspase-12, as well as upregulation of Bcl-2 during HR. Furthermore, FABP4 silencing activated the PI3K/Akt pathway. Blocking this pathway by the specific PI3K inhibitor-LY294002 restored HR-induced ER stress and subsequently reversed the protective effect of FABP4 silencing on HR injury. Taken together, our findings revealed that FABP4 silencing exerts protective effects against HR injury in H9c2 cells through inhibiting ER stress-induced cell apoptosis via activation of the PI3K/Akt pathway.
    Keywords:  Apoptosis; Endoplasmic reticulum stress; FABP4; Hypoxia reoxygenation; PI3K/Akt
    DOI:  https://doi.org/10.1016/j.lfs.2019.03.046
  48. Cancer Cell Int. 2019 ;19 60
    Song J, Zhao W, Lu C, Shao X.
      Background: Effective therapy for hepatocellular carcinoma (HCC) is currently an imperative issue, and sorafenib is a first-line drug for the treatment of HCC. However, the clinical benefit of sorafenib is often impaired by drug resistance. Accordingly, the present study was conducted to investigate the molecular mechanisms involving sorafenib resistance, with a focus on large tumor suppressor 2 (LATS2) and mitophagy.Methods: HepG2 liver cancer cells were treated with sorafenib and infected with adenovirus-loaded LATS2 (Ad-LATS2). Cell death, proliferation and migration were measured via western blotting analysis, immunofluorescence and qPCR. Mitochondrial function and mitophagy were determined via western blotting and immunofluorescence.
    Results: Our data indicated that LATS2 expression was repressed by sorafenib treatment, and overexpression of LATS2 could further enhance sorafenib-mediated apoptosis in HepG2 liver cancer cells. At the molecular level, mitochondrial stress was triggered by sorafenib treatment, as evidenced by decreased mitochondrial membrane potential, increased mitochondrial ROS production, more cyc-c release into the nucleus, and elevated mitochondrial pro-apoptotic proteins. However, in response to mitochondrial damage, mitophagy was activated by sorafenib treatment, whereas LATS2 overexpression effectively inhibited mitophagy activity and thus augmented sorafenib-mediated mitochondrial stress. Subsequently, we also demonstrated that the AMPK-MFN2 signaling pathway was involved in mitophagy regulation after exposure to sorafenib treatment and/or LATS2 overexpression. Inhibition of the AMPK pathway interrupted mitophagy and thus enhanced the antitumor property of sorafenib, similar to the results obtained via overexpression of LATS2.
    Conclusions: Altogether, our findings revealed the importance of the LATS2/AMPK/MFN2/mitophagy axis in understanding sorafenib resistance mechanisms, with a potential application to increase the sensitivity response of sorafenib in the treatment of liver cancer.
    Keywords:  AMPK pathway; HCC; LATS2; Mitophagy; Sorafenib
    DOI:  https://doi.org/10.1186/s12935-019-0778-1
  49. Biomed Pharmacother. 2019 Mar 26. pii: S0753-3322(18)36525-9. [Epub ahead of print]114 108732
    El-Magd MA, Mohamed Y, El-Shetry ES, Elsayed SA, Abo Gazia M, Abdel-Aleem GA, Shafik NM, Abdo WS, El-Desouki NI, Basyony MA.
      Pretreatment of mesenchymal stem cells (MSCs) with melatonin (Mel) improves their potential therapeutic effect on chronic diseases and cancers. However, this preconditioning strategy may direct the effect of Mel toward MSCs alone and deprive cancer cells of the oncostatic effect of Mel. Herein, we hypothesized that Mel given before transplantation of non-preconditioned MSCs may maximize the therapeutic outcome via the oncostatic effect of Mel by preparing a suitable tumor microenvironment for MSCs. Female rats (n = 60) were equally divided into 6 groups; normal control, diethylnitrosamine (DEN), DEN + Mel, DEN + MSCs, DEN + MSCs preconditioned with Mel, and DEN + MSCs + Mel. The obtained data revealed that administration of Mel before MSCs treatment without preconditioning yielded a better ameliorative effect against DEN-induced hepatocellular carcinoma (HCC) as evidenced by: 1) reduced serum levels of alpha fetoprotein and gamma-glutamyl transferase; 2) decreased number and area of glutathione S-transferase placental positive foci; 3) induced apoptosis (as indicated by increased cleaved caspase-3 activity, upregulated expression of proapoptotic genes Bax and caspase 3 and downregulated expression of anti-apoptotic genes Bcl2, survivin); 4) decreased malondialdehyde level and increased activities of superoxide dismutase, catalase, and glutathione peroxidase enzymes; and 5) reduced inflammation, angiogenesis and metastasis as indicated by downregulated expression of interleukin 1 beta, nuclear factor kappa B, vascular endothelial growth factor, and matrix metallopeptidase 9 genes and upregulated expression of metalloproteinase inhibitor 1 gene. Thus, administration of Mel before MSCs (without preconditioning) fostered the survival and therapeutic potential of MSCs in HCC, possibly through induction of apoptosis and inhibition of inflammation and oxidative stress. This new strategy showed better therapeutic outcomes and may improve MSC-based therapies for HCC.
    Keywords:  Apoptosis; Hepatocellular carcinoma; Melatonin; Mesenchymal stem cells; Oxidative stress
    DOI:  https://doi.org/10.1016/j.biopha.2019.108732
  50. Environ Toxicol Chem. 2019 Mar 28.
    Kumar N, Awoyemi O, Willis A, Schmitt C, Ramalingam L, Moustaid-Moussa N, Crago J.
      Azole fungicides have entered the aquatic environment through agricultural and residential runoff. In the present study, we compared the off-target toxicity of tebuconazole, propiconazole, and myclobutanil using embryo-larval zebrafish as a model. The aim of this study was to investigate a relative toxicity of tebuconazole, propiconazole, and myclobutanil using multiple level endpoints such as behavioral endpoints and enzymatic and molecular biomarkers associated with their mode of action. Zebrafish embryos were exposed to azoles at environmentally relevant and high concentrations, 0.3, 1.0 and 1000 µg/L, starting at 5 hours post- fertilization (hpf) up to 48 hpf, as well as 5 days post fertilization (dpf). Relative mRNA expressions of CYP51, GST, Casp9, p53, and BAX were measured to assess toxicity due to fungicides at mRNA level, whereas caspase 3/7 (apoptosis) and MDA (lipid peroxidation) levels were measured at enzymatic level. Furthermore, mitochondrial dysfunction was measure through the Mito Stress test using the Seahorse XFe24 at 48 hpf. Additionally, light: dark movement behavior was monitored at 5 dpf using Danio Vision® to understand adverse effects at the organismal level. There was no significant difference in the light: dark behavior with exposure to azoles as compared to controls. The molecular biomarkers indicated that propiconazole and myclobutanil induced lipid peroxidation, oxidative stress, and potentially apoptosis at environmentally relevant concentrations (0.3 and 1 µg/L). The results from the mitochondrial respiration assay indicated a slight decrease in spare respiratory capacity with an acute exposure (48 hpf) to all three azoles at 1000 µg/L. Based on our results, propiconazole and myclobutanil are acutely toxic as compared to tebuconazole in aquatic organisms at environmentally relevant concentrations. This article is protected by copyright. All rights reserved.
    Keywords:  Apoptosis; Lipid peroxidation; Myclobutanil; Propiconazole; Tebuconazole; Zebrafish
    DOI:  https://doi.org/10.1002/etc.4429
  51. J Ethnopharmacol. 2019 Mar 26. pii: S0378-8741(18)33248-3. [Epub ahead of print]
    Rezaee-Khorasany A, Razavi BM, Taghiabadi E, Yazdi AT, Hosseinzadeh H.
      ETHNOPHARMACOLOGICAL RELEVANCE: Saffron (Crocus sativus L.) is considered in the Iranian traditional medicine because of many therapeutic properties such as sedative agent, strengthen the stomach and liver, improving the uterus disorders and infectious wounds. The detoxification of alcohol was one of the most important of saffron effects in ancient medicine.AIM OF THE STUDY: In the current research, the protective effects of saffron aqueous extract (Aq. Ext.) versus oxidative stress, apoptosis, inflammation, histopathological and biochemical abnormalities induced by ethanol were evaluated.
    MATERIALS & METHODS: The male Wistar rats were divided into seven groups consisted of 6 rats in control (distilled water), ethanol (5 g/kg - 50% v/v), Aq. Ext. (40, 80 and 160 mg/kg) plus ethanol, Aq. Ext. 80 and 160 mg/kg. Animals were treated for four weeks and at the end of treatment period, histopathological damages, biochemical markers, apoptosis, levels of MDA and GSH, TNF-α and IL-6 were evaluated.
    RESULTS: Ethanol induced nephrotoxicity and hepatotoxicity as evidenced by histopathological damages and biochemical abnormalities. The level of MDA was significantly enhanced while GSH content was remarkably reduced in ethanol-treated rats, but protective groups restored them. Also, the levels of TNF-α and IL-6 were regulated by Aq. Ext. Furthermore, the effects of ethanol on histopathological and biochemical parameters were improved by Aq. Ext. The ethanol increased the expression of Bax/Bcl2 ratio, caspase-3, -8, and -9. Real-time PCR and western blot analysis proved that Aq. Ext. treatment inhibited apoptosis induced by ethanol through decreasing the Bax/Bcl2 ratio (mRNA and protein) and caspases-3, -8, and -9 levels in the kidney and liver.
    CONCLUSION: The results of this research demonstrated that Aq. Ext. could exert protective effects against ethanol toxicity in rat kidney and liver via antioxidant, anti-apoptosis, and anti-inflammatory effects.
    Keywords:  Apoptosis; Ethanol; Hepatotoxicity; Nephrotoxicity; Oxidative stress; Saffron
    DOI:  https://doi.org/10.1016/j.jep.2019.03.048
  52. Phytochemistry. 2019 Mar 26. pii: S0031-9422(18)30720-9. [Epub ahead of print]162 173-182
    Li Y, Sun Y, Zhu M, Zhu R, Zhang J, Zhou J, Wang T, Qiao Y, Lou H.
      Eight previously undescribed sacculatane diterpenoids, epiphyllins A-H, and one unknown bibenzyl-based isopentene along with seven known compounds were isolated from the Chinese liverwort Pellia epiphylla (L.) Corda. Their structures were established unequivocally on the basis of spectroscopic data and CD measurement. The quinine reductase-inducing activity evaluation demonstrated that epiphyllins A-D, 1β-hydroxysacculatanolide and pellianolactone B displayed moderate antioxidant effect. Further investigation of pellianolactone B revealed its protective effects on H2O2-induced oxidative insults and apoptosis in PC12 cells.
    Keywords:  Apoptosis; Epiphyllins A−H; Liverworts; PC12 cells; Pellia epiphylla (L.) Corda; Pelliaceae; Sacculatane diterpenoids
    DOI:  https://doi.org/10.1016/j.phytochem.2019.03.007
  53. Free Radic Biol Med. 2019 Mar 26. pii: S0891-5849(19)30264-3. [Epub ahead of print]
    Chen C, Zhou Y, Hu C, Wang Y, Yan Z, Li Z, Wu R.
      Endometriosis is associated with inflammatory reaction, and reactive oxidative species (ROS) are highly pro-inflammatory factors. Mitochondria are responsible for the production of ROS and energy. However, little is known about how mitochondria regulate ROS generation and energy metabolism in endometriosis. In our study, we investigated mitochondrial structure and function of ectopic endometrial stromal cells (ESCs) in ovarian endometriosis. We found mitochondria in ectopic ESCs generated more ROS and energy than controlled groups. Mitochondrial superoxide dismutase (SOD2), as an antioxidant enzyme, was found highly expressed in ectopic endometrium compared with normal endometrium. Due to its antioxidant role, SOD2 promoted the development of endometriosis by maintaining functional mitochondria to support high energetic metabolism of ectopic ESCs. We also showed that SOD2 promoted cell proliferation and migration in ovarian endometriosis. Inhibiting SOD2 expression reduced proliferation and migration of ectopic ESCS, and increased cell apoptosis. Therefore, understanding the role of mitochondrial dysfunction and SOD2 in ovarian endometriosis may provide new strategies to treat this disease.
    Keywords:  Mitochondria; Mitochondrial superoxide dismutase; Ovarian endometriosis; Oxidative stress; Reactive oxidative species
    DOI:  https://doi.org/10.1016/j.freeradbiomed.2019.03.027
  54. Aging (Albany NY). 2019 Mar 26.
    Zhang Y, Gao J, Sun W, Wen X, Xi Y, Wang Y, Wei C, Xu C, Li H.
      Hydrogen sulfide (H2S) reduces ischemia/reperfusion (I/R) injury and apoptosis and restores the cardioprotective effects of ischemic post-conditioning (PC) in aged cardiomyocytes by inhibiting oxidative stress and endoplasmic reticulum stress and increasing autophagy. However, the mechanism is unclear. In the present study, we observed a loss of PC-mediated cardioprotection of aged cardiomyocytes. NaHS (a H2S donor) exerted significant protective effects against H/R-induced cell damage, apoptosis, production of cleaved caspase-3 and caspase-9, and release of cytochrome c. NaHS also reversed the H/R-induced reduction in cell viability and increased HB-EGF expression, cellular HB-EGF content, and EGFR phosphorylation. Additionally, NaHS increased expression of Bcl-2, c-myc, c-fos and c-jun, and the phosphorylation of ERK1/2, PI3K, Akt and GSK-3β. PC alone did not provide protection to H/R-treated aged cardiomyocytes, but it was significantly restored by supplementation of NaHS. The beneficial effects of NaHS during PC were inhibited by EGFR knockdown, AG1478 (EGFR inhibitor), PD98059 (ERK1/2 inhibitor) or LY294002 (PI3K inhibitor). These results suggest that exogenous H2S restores PC-mediated cardioprotection by up-regulating HB-EGF/EGFR signaling, which activates the ERK1/2-c-myc (and fos and c-jun) and PI3K-Akt- GSK-3β pathways in the aged cardiomyocytes.
    Keywords:  H9C2 cells; HB-EGF/EGFR pathway; aged cardiomyocytes; hydrogen sulfide; post-conditioning
    DOI:  https://doi.org/10.18632/aging.101866
  55. J Cell Mol Med. 2019 Mar 28.
    Zhang T, Zhu X, Wu H, Jiang K, Zhao G, Shaukat A, Deng G, Qiu C.
      It is well established that cancer cells depend upon aerobic glycolysis to provide the energy they need to survive and proliferate. However, anti-glycolytic agents have yielded few positive results in human patients, in part due to dose-limiting side effects. Here, we discovered the unexpected anti-cancer efficacy of Polydatin (PD) combined with 2-deoxy-D-glucose (2-DG), which is a compound that inhibits glycolysis. We demonstrated in two breast cell lines (MCF-7 and 4T1) that combination treatment with PD and 2-DG induced cell apoptosis and inhibited cell proliferation, migration and invasion. Furthermore, we determined the mechanism of PD in synergy with 2-DG, which decreased the intracellular reactive oxygen (ROS) levels and suppressed the PI3K/AKT pathway. In addition, the combined treatment inhibited the glycolytic phenotype through reducing the expression of HK2. HK2 deletion in breast cancer cells thus improved the anti-cancer activity of 2-DG. The combination treatment also resulted in significant tumour regression in the absence of significant morphologic changes in the heart, liver or kidney in vivo. In summary, our study demonstrates that PD synergised with 2-DG to enhance its anti-cancer efficacy by inhibiting the ROS/PI3K/AKT/HIF-1α/HK2 signalling axis, providing a potential anti-cancer strategy.
    Keywords:  2-deoxy-D-glucose; PI3K/AKT; Polydatin; breast cancer; glycolysis
    DOI:  https://doi.org/10.1111/jcmm.14276
  56. Aging Cell. 2019 Mar 24. e12951
    Chen L, Yang R, Qiao W, Zhang W, Chen J, Mao L, Goltzman D, Miao D.
      We tested the hypothesis that 1,25-dihydroxyvitamin D3 [1α,25(OH)2 D3 ] has antiaging effects via upregulating nuclear factor (erythroid-derived 2)-like 2 (Nrf2), reducing reactive oxygen species (ROS), decreasing DNA damage, reducing p16/Rb and p53/p21 signaling, increasing cell proliferation, and reducing cellular senescence and the senescence-associated secretory phenotype (SASP). We demonstrated that 1,25(OH)2 D3 -deficient [1α(OH)ase-/- ] mice survived on average for only 3 months. Increased tissue oxidative stress and DNA damage, downregulated Bmi1 and upregulated p16, p53 and p21 expression levels, reduced cell proliferation, and induced cell senescence and the senescence-associated secretory phenotype (SASP) were observed. Supplementation of 1α(OH)ase-/- mice with dietary calcium and phosphate, which normalized serum calcium and phosphorus, prolonged their average lifespan to more than 8 months with reduced oxidative stress and cellular senescence and SASP. However, supplementation with exogenous 1,25(OH)2 D3 or with combined calcium/phosphate and the antioxidant N-acetyl-l-cysteine prolonged their average lifespan to more than 16 months and nearly 14 months, respectively, largely rescuing the aging phenotypes. We demonstrated that 1,25(OH)2 D3 exerted an antioxidant role by transcriptional regulation of Nrf2 via the vitamin D receptor (VDR). Homozygous ablation of p16 or heterozygous ablation of p53 prolonged the average lifespan of 1α(OH)ase-/- mice on the normal diet from 3 to 6 months by enhancing cell proliferative ability and reducing cell senescence or apoptosis. This study suggests that 1,25(OH)2 D3 plays a role in delaying aging by upregulating Nrf2, inhibiting oxidative stress and DNA damage,inactivating p53-p21 and p16-Rb signaling pathways, and inhibiting cell senescence and SASP.
    Keywords:  Nrf2; aging; cell senescence; p16 and p53; vitamin D
    DOI:  https://doi.org/10.1111/acel.12951
  57. Molecules. 2019 Mar 26. pii: E1184. [Epub ahead of print]24(6):
    Guan P, Sun ZM, Luo LF, Zhao YS, Yang SC, Yu FY, Wang N, Ji ES.
      Iron-induced oxidative stress has been found to be a central player in the pathogenesis of kidney injury. Recent studies have indicated H₂ can be used as a novel antioxidant to protect cells. The present study was designed to investigate the protective effects of H₂ against chronic intermittent hypoxia (CIH)-induced renal injury and its correlation mechanism involved in iron metabolism. We found that CIH-induced renal iron overloaded along with increased apoptosis and oxidative stress. Iron accumulates mainly occurred in the proximal tubule epithelial cells of rats as showed by Perl's stain. Moreover, we found that CIH could promote renal transferrin receptor and divalent metal transporter-1 expression, inhibit ceruloplasmin expression. Renal injury, apoptosis and oxidative stress induced by CIH were strikingly attenuated in H₂ treated rats. In conclusion, hydrogen may attenuate CIH-induced renal injury at least partially via inhibiting renal iron overload.
    Keywords:  chronic intermittent hypoxia; hydrogen; iron; kidney
    DOI:  https://doi.org/10.3390/molecules24061184
  58. Redox Biol. 2019 Mar 08. pii: S2213-2317(19)30028-X. [Epub ahead of print]24 101153
    Wang X, Gao M, Zhang J, Ma Y, Qu W, Liang J, Wu H, Wen H.
      The early immature CD34+ acute myeloid leukemia (AML) cell subpopulation-acute myeloid leukemia progenitor cells (APCs), is often resistant to conventional chemotherapy, making them largely responsible for the relapse of AML. However, to date, the eradication of APCs remains a major challenge. We previously reported a naturally occurring secolignan- Peperomin E (PepE) and its analog 6-methyl (hydroxyethyl) amino-2, 6-dihydropeperomin E (DMAPE) that selectively target and induce oxidative stress-mediated apoptosis in KG-1a CD34+ cells (an APCs-like cell line) in vitro. We therefore further evaluated the efficacy and the mechanism of action of these compounds in this study. We found that PepE and DMAPE have similar potential to eliminate primary APCs, with no substantial toxicities to the normal cells in vitro and in vivo. Mechanistically, these agents selectively inhibit TrxR1, an antioxidant enzyme aberrantly expressed in APCs, by covalently binding to its selenocysteine residue at the C-terminal redox center. TrxR1 inhibition mediated by PepE (DMAPE) leads to the formation of cellular selenium compromised thioredoxin reductase-derived apoptotic protein (SecTRAP), oxidation of Trx, induction of oxidative stress and finally activation of apoptosis of APCs. Our results demonstrate a potential anti-APCs molecular target - TrxR1 and provide valuable insights into the mechanism underlying PepE (DMAPE)-induced cytotoxicity of APCs, and support the further preclinical investigations on PepE (DMAPE)-related therapies for the treatment of relapsed AML.
    Keywords:  Acute myeloid leukemia progenitor cells; Orally bioavailable analog; Oxidative-mediated apoptosis; Peperomin E; Thioredoxin reductase 1
    DOI:  https://doi.org/10.1016/j.redox.2019.101153
  59. Small. 2019 Mar 26. e1900011
    Wang H, Lin Q, Yin L, Yang Y, Qiu Y, Lu C, Yang H.
      Organic framework polymers have attracted much interest due to the enormous potential design space offered by the atomically precise spatial assembly of organic molecular building blocks. The morphology control of organic frameworks is a complex issue that hinders the development of organic frameworks for practical applications. Biomimetic self-assembly is a promising approach for designing and fabricating multiple-functional nanoarchitectures. A bioinspired hollow flower-like organic framework nanosphere heterostructure comprised of carbon nitride and polydopamine (g-C3N4@PDA) is successfully synthesized via a mild and green method. This heterostructure can effectively avoid the agglomeration of nanosheets to better access the hollow nanospheres with high open-up specific surface area. The electron delocalization of g-C3N4 and PDA under visible light can largely promote photoelectron transfer and enhance the photocatalytic activity of the g-C3N4@PDA. Furthermore, the g-C3N4@PDA can effectively enhance the generation of reactive oxygen species under irradiation, which can lead to cell apoptosis and enhance the performance for cancer therapy. Therefore, the as-prepared g-C3N4@PDA provides a paradigm of highly efficient photocatalyst that can be used as nanomedicine toward cancer therapy. This study could open up a new avenue for exploiting more other potential hollow nanosphere organic frameworks.
    Keywords:  carbon nitride; organic frameworks; photocatalysis; photodynamic therapy; polydopamine
    DOI:  https://doi.org/10.1002/smll.201900011
  60. Cell Death Dis. 2019 Mar 27. 10(4): 289
    Li Y, Qin Y, Yang C, Zhang H, Li Y, Wu B, Huang J, Zhou X, Huang B, Yang K, Wu G.
      The authors have retracted this article [1] because of numerous errors in the figures. Specifically.
    DOI:  https://doi.org/10.1038/s41419-019-1482-8
  61. Eur J Med Chem. 2019 Mar 17. pii: S0223-5234(19)30245-4. [Epub ahead of print]171 180-194
    Mrozek-Wilczkiewicz A, Malarz K, Rejmund M, Polanski J, Musiol R.
      Thiosemicarbazones (TSC) are a subclass of iron-chelating agents that are believed to have an anticancer activity. The high potential for the application of this compound class can be illustrated by a fact that three TSC have entered clinical trials. The ability to chelate metal ions results in several biochemical changes in the cellular metabolism and growth. An important factor that determines the antitumor activity of TSC is a level of iron regulatory proteins and the antioxidant potential that is specific for each type of cancer cell. However, despite the increasing interest in TSC, their mechanism of anticancer activity is still unclear. For a more effective and rational design, it is crucial to determine and describe the abovementioned issues. In this report, we describe a series of new TSC that are designed on the four main structural scaffolds. The anticancer activity of these compounds was evaluated against a panel of cancer cell lines including colon and breast cancers and gliomas. Special attention was paid to the metal-dependent proteins. The impact of the tested TSC on the cell cycle and redox homeostasis was also determined. These results confirm a p53-independent mechanism of apoptosis.
    Keywords:  Anticancer activity; Apoptosis; Cell cycle inhibition; Iron chelators; Reactive oxygen species; Thiosemicarbazones
    DOI:  https://doi.org/10.1016/j.ejmech.2019.03.027
  62. Neurotoxicology. 2019 Mar 20. pii: S0161-813X(18)30373-5. [Epub ahead of print]
    Sun C, Mo M, Wang Y, Yu W, Song C, Wang X, Chen S, Liu Y.
      This study investigated the expression and role of immunoproteasome (i-proteasome) in a cell model of Parkinson's disease (PD). The cytotoxicity of rotenone was measured by CCK-8 assay. The i-proteasome β1i subunit PSMB9 was suppressed by a specific shRNA or transfected with an overexpression plasmid in the SH-SY5Y cells. Under the exposure to rotenone or not, the expression of constitutive proteasome β subunits, i-proteasome βi subunits, antigen presentation related proteins, α-syn and TH were detected by Western blot in PSMB9-silenced or -overexpressed cells, and the proteasomal activities were detected by fluorogenic peptide substrates. The location of i-proteasome βi subunits and α-syn were detected by immunofluorescence staining. The levels of ROS, GSH and MDA were measured by commercial kits. Cell apoptosis was detected by flow cytometry. Besides impairing the constitutive proteasomes, rotenone induced the expression of βi subunits of i-proteasome and antigen presentation related proteins such as TAP1, TAP2 and MHC-I. Silencing or overexpressing PSMB9 had no obvious effect on the levels of other subunits, but could regulate the chymotrypsin-like activity of 20S proteasome and the expression of TAP1, TAP2 and MHC-I. Three βi subunits (PSMB9, PSMB10, PSMB8) of i-proteasome were all co-localized with α-syn. PSMB9 knockdown aggravated accumulation of α-syn, degradation of TH, release of ROS, increased level of MDA, decreased level of GSH and eventually promoted apoptosis in SH-SY5Y cells after rotenone treatment, while over-expression of PSMB9 could attenuate these toxic effects of rotenone. I-proteasome is activated in SH-SY5Y cells treated with rotenone and may play a neuroprotective role.
    Keywords:  Immunoproteasome; PSMB9; Parkinson’s disease; Rotenone
    DOI:  https://doi.org/10.1016/j.neuro.2019.03.004
  63. Can J Physiol Pharmacol. 2019 Mar 27. 1-11
    El-Sayed RM, Ahmed HI, Abd El-Lateef AES, Ali AA.
      Hepatic injury is one of the most common complications associated with cisplatin (CIS) use. Recently, liver protection lines are being discovered to stop the hepatic cell death due to inflammatory and apoptotic perturbations. l-arginine has protective effects in several models of liver injury. This study was designed to investigate the possible protective effect of l-arginine against CIS-induced acute hepatic injury in rats. Rats were divided into 4 groups: control, l-arginine, CIS, l-arginine + CIS. Liver function, oxidative stress, inflammatory cytokines, and apoptosis markers were assessed. l-arginine pretreatment protected the liver against CIS-induced toxicity as indicated by significantly alleviating the changes in liver function along with restoration of the antioxidant status. This finding was confirmed with the markedly improved pathological changes. l-arginine showed anti-inflammatory effect through the reduction of liver expression of iNOS, TNF-α, and NF-κβ, which were ameliorated to significant levels. Furthermore, l-arginine administration downregulated the liver expression of the apoptotic marker, caspase-3. The results recommend l-arginine as a hepatoprotective agent against CIS toxicity. Mostly, this hepatoprotective effect of l-arginine involved anti-inflammatory and anti-apoptotic activities.
    Keywords:  -arginine; NF-κβ; TNF-α; apoptose; apoptosis; cisplatin; cisplatine; hepatic injury; iNOS; lésions hépatiques
    DOI:  https://doi.org/10.1139/cjpp-2018-0706
  64. Nano Lett. 2019 Mar 25.
    Zhang K, Tu M, Gao W, Cai X, Song F, Chen Z, Zhang Q, Wang J, Jin C, Shi J, Yang X, Zhu Y, Gu W, Hu B, Zheng Y, Zhang H, Tian M.
      Ischemic stroke is a devastating disease and one of the leading causes of mortality worldwide. Overproduction of reactive oxygen and nitrogen species (RONS) following ischemic insult is known as a key factor in exacerbating brain damage. Thus, RONS scavengers that can block excessive production of RONS have great therapeutic potential. Herein, we propose an efficient treatment strategy in which an artificial nanozyme with multienzyme activity drives neuroprotection against ischemic stroke primarily by scavenging RONS. Specifically, through a facile, Bi3+-assisted, template-free synthetic strategy, we developed hollow Prussian blue nanozymes (HPBZs) with multienzyme activity to scavenge RONS in a rat model of ischemic stroke. The comprehensive characteristics of HPBZs against RONS were explored. Apart from attenuating oxidative stress, HPBZs also suppressed apoptosis and counteracted inflammation both in vitro and in vivo, thereby contributing to increased brain tolerance of ischemic injury with minimal side effects. This study provides a proof of concept for a novel class of neuroprotective nanoagents that might be beneficial for treatment of ischemic stroke and other RONS-related disorders.
    DOI:  https://doi.org/10.1021/acs.nanolett.8b04729
  65. J Trace Elem Med Biol. 2019 May;pii: S0946-672X(18)30630-8. [Epub ahead of print]53 1-7
    Cengiz M, Cetik Yildiz S, Demir C, Şahin İK, Teksoy Ö, Ayhanci A.
      This study aims to examine cyclophosphamide (CP) exsposure associated toxicity on rat livers and the likely defensive effects of boric acid (BA). The rats used in this study were divided into four groups: control group, CP group, BA group, and BA + CP group. The present study was carried out using routine histological H&E stain, immunohistochemical stain caspase-3 as apoptotic marker, serum biochemical analysis for liver function markers (alanine transaminase (ALT), aspartate transaminase (AST) and alkalen phosphatase (ALP)), oxidative stress markers (total oxidant status (TOS), oxidative stress index (OSI) and total antioxidant capacity marker (TAC)). In the CP group, the levels of ALT, AST, ALP, TOS, OSI and caspase-3 increased whereas TAC levels decreased compared with the control group. In the BA + CP group, the levels of ALT, AST, ALP, TOS, OSI and caspase-3 decreased whereas TAC levels increased compared with the CP group. The histopathological evaluation of light microscope images and immunohistochemical caspase-3 activity in the BA + CP group were found to be decrease compared with those in the CP group. In conclusion, BA was successful in defending the liver against apoptosis and histopathological changes that are attributable to CP.
    Keywords:  Apoptosis; Boric acid; Cyclophosphamide; Liver injury; Oxidative stress
    DOI:  https://doi.org/10.1016/j.jtemb.2019.01.013
  66. J Trace Elem Med Biol. 2019 May;pii: S0946-672X(18)30504-2. [Epub ahead of print]53 34-40
    Soares ATG, Rodrigues LBL, Salgueiro WG, Dal Forno AHC, Rodrigues CF, Sacramento M, Franco J, Alves D, Oliveira RP, Pinton S, Ávila DS.
      Organic selenium compounds have several pharmacological activities already described, as anti-inflammatory and antitumor activities, which have been attributed to their antioxidant effects. Because they are promising in pharmacology, the synthesis of these compounds has increased significantly. As many new molecules are synthesized the use of a simple model like Caenorhabditis elegans is highly advantageous for initial evaluation of the toxicity and therapeutic potential of these molecules. The objective of this study was to evaluate the toxicity and antioxidant capacity of a series of selenotriazoles compounds in C. elegans. The animals were exposed to the compounds in liquid medium for only 30 min at the first larval stage (L1). The compounds had no toxic effects at the concentrations tested. Treatment with selenotriazoles (10 μM) partially reversed the stress induced by the pesticide paraquat (1 mM). Se-Tz Ia compound partially increased the survival of worms treated with H2O2 (0.5 mM). The compounds also increased the longevity of mev-1 mutants, which have a reduced life span by the production of excessive reactive oxygen species (ROS) in the mitochondria caused by a mutation in complex II of the electron transport chain. In addition, the compounds reduced the levels of ROS determined by the fluorescent probe DCF-DA as well as also reduced catalase enzyme activity in these animals. Based on the results found, it is possible to conclude that the compounds have antioxidant activity mainly in oxidative stress condition generated by a mitochondrial dysfunction in C. elegans.
    Keywords:  Caenorhabditis elegans; Mitochondria; Oxidative stress; Selenium; Selenotriazoles
    DOI:  https://doi.org/10.1016/j.jtemb.2019.01.017
  67. Dev Comp Immunol. 2019 Mar 22. pii: S0145-305X(18)30561-5. [Epub ahead of print]97 28-37
    Dai YJ, Cao XF, Zhang DD, Li XF, Liu WB, Jiang GZ.
      The aim of this article is to investigate the mechanism of lipotoxicity induced by high-fat diets (HFD) in Megalobrama amblycephala. In the present study, fish (average initial weight 40.0 ± 0.35 g) were fed with two fat levels (6% and 11%) diets with four replicates for 60 days. At the end of the feeding trial, fish were challenged by thioacetamide (TAA) and survival rate was recorded for the next 96 h. The result showed that long-term HFD feeding induced a significant increase (P < 0.05) in the levels of aspartate aminotransferase (GOT) and alanine aminotransferase (GPT) in plasma. In addition, liver histopathological analysis showed an increased dilation of the blood vessels, erythrocytes outside of the blood vessels and vacuolization in fish fed with high-fat diet. After TAA challenge, compared with group fed with normal-fat diets (NFD), fish fed with HFD showed a significantly (P < 0.05) low survival rate. After feeding Megalobrama amblycephala with HFD for 60 days, the protein content and gene expression of pro-inflammatory factors were significantly elevated (P < 0.05). The protein and gene relative expressions of a Caspase-3, Caspase-9 and CD68 were significantly increased (P < 0.05), while antioxidant-related enzyme activities were significantly reduced (P < 0.05) in the liver of fish fed with HFD. In addition, HFD feeding also induced genotoxicity. Comet assay showed a significantly (P < 0.05) elevated DNA damage in blunt snout bream fed with HFD. Compared with normal-fat diets (NFD) group, the protein expression of γH2AX and gene expressions involved in cell cycle arrest were significantly increased (P < 0.05) in fish fed with HFD. Data in this research showed that lipotoxicity induced by HFD was likely mediated by chronic inflammation regulating macrophage recruitment, apoptosis and DNA damage. The study was valuable to understand the mechanism by which liver injury is induced in fish fed with HFD.
    Keywords:  Apoptosis; DNA damage; High-fat diets; Inflammation; Liver injury; Oxidative stress
    DOI:  https://doi.org/10.1016/j.dci.2019.03.009
  68. J Photochem Photobiol B. 2019 Mar 14. pii: S1011-1344(18)31310-1. [Epub ahead of print]194 32-45
    Antunovic M, Matic I, Nagy B, Caput Mihalic K, Skelin J, Stambuk J, Josipovic P, Dzinic T, Paradzik M, Marijanovic I.
      Sun or therapy-related ultraviolet B (UVB) irradiation induces different cell death modalities such as apoptosis, necrosis/necroptosis and autophagy. Understanding of mechanisms implicated in regulation and execution of cell death program is imperative for prevention and treatment of skin diseases. An essential component of death-inducing complex is Fas-associated protein with death domain (FADD), involved in conduction of death signals of different death modalities. The purpose of this study was to enlighten the role of FADD in the selection of cell death mode after narrow-band UVB (NB-UVB) irradiation using specific cell death inhibitors (carbobenzoxy-valyl-alanyl-aspartyl-[O-methyl]- fluoromethylketone (zVAD-fmk), Necrostatin-1 and 3-Methyladenine) and FADD-deficient (FADD-/-) mouse embryonic fibroblasts (MEFs) and their wild type (wt) counterparts. The results imply that lack of FADD sensitized MEFs to induction of receptor-interacting protein 1 (RIPK1)-dependent apoptosis by the generation of reactive oxygen species (ROS), but without activation of the proteins p53, Bax and Bcl-2 as well as without the enrolment of calpain-2. Autophagy was established as a contributing factor to NB-UVB-induced death execution. By contrast, wt cells triggered intrinsic apoptotic pathway that was resistant to the inhibition by zVAD-fmk and Necrostatin-1 pointing to the mechanism overcoming the cell survival. These findings support the role of FADD in prevention of autophagy-dependent apoptosis.
    Keywords:  Autophagy; Cell death inhibitors; FADD; NB-UVB irradiation; RIPK1-dependent apoptosis
    DOI:  https://doi.org/10.1016/j.jphotobiol.2019.03.007
  69. Nanotechnology. 2019 Mar 25.
    Zhang M, Zhao L, Du F, Wu Y, Cai R, Xu L, Jin H, Zou S, Gong A, Du F.
      Excessive reactive oxygen species can lead to irreversible damage for human body in vivo, therefore it is highly desirable to exploit efficient antioxidant. Recently, cerium oxide nanoparticles have attracted extensive attention in the field of biomedicine due to their excellent antioxidant properties. In this study, Cerium-doped carbon quantum dots (Ce-doped CQDs) with hydroxyl radical scavenging capacity were synthesized for first time by one-step hydrothermal carbonization method. The resultant Ce-doped CQDs with the average particle size of 2.5 nm possessed properties of good water solubility, colloid stability and strong fluorescence, which are similar to traditional carbon quantum dots. Meanwhile, Ce-doped CQDs had good biocompatibility and negligible cytotoxicity. Taking advantage of inherent ultra-small size, Ce-doped CQDs exhibited highly Ce3+/Ce4+ ratio at the surface of particles. The radical scavenging capability of Ce-doped CQDs was proved by a simple photometric system in vitro, which provided direct evidence for its antioxidant potency. Furthermore, Ce-doped CQDs had a high ability to protect cells from hydrogen peroxide-induced damage by scavenging hydroxyl radicals. These results suggest that Ce-doped CQDs as a new ROS scavenger may provide potential prospects for the treatment of oxidative stress-related diseases.
    Keywords:  Antioxidant; Carbon quantum dots; Cerium; Radical Scavenging
    DOI:  https://doi.org/10.1088/1361-6528/ab12ef
  70. J Ethnopharmacol. 2019 Mar 21. pii: S0378-8741(18)30212-5. [Epub ahead of print]237 108-115
    Murugesan S, Pandiyan A, Saravanakumar L, Moodley K, Mackraj I.
      ETHNOPHARMACOLOGICAL RELEVANCE: Tulbaghia violacea Harv. (TVL) is a folk medicine, native to South Africa which has previously shown antioxidant, anti-hypertensive and anti-diabetic effects.THE AIM OF THE STUDY: The aim of the current study was to investigate the protective role of wild garlic or TVL on isoproterenol (ISO)-induced myocardial necrosis in rats.
    MATERIALS AND METHODS: Animal (n = 6 each group) were pre and co-treated with TVL (60 mg/kg body weight) daily for 30 days. Myocardial necrosis was administrated by subcutaneous injection of ISO (85 mg/kg body weight) into rats on 29th and 30th day. On the 31st day, rats were anaesthetized and blood, heart samples were obtained for the biochemical, histopathological and molecular study. The specific protein target analysis from TVL was done by reverse docking study (reverse pharmacophore mapping) using PharmMapper.
    RESULTS: The levels of cardiac markers, lipid peroxidation products, and heart rate were considerably increased in ISO-induced myocardial necrosis in rats whilst plasma enzymatic antioxidants were significantly decreased. Myocardial necrotic mRNA genes were increased in ISO-induced myocardial necrosis in rats compared to controls. Pre and co-treatment with TVL and ramipril of myocardial necrosis in rats showed significant effects on all the biochemical and molecular studies evaluated. TVL reduced heart rate, prevented oxidative stress and downregulated the Fas-receptor and caspase-mediated apoptosis-signaling pathway, and heart muscle damage in myocardial necrosis in rats. The specific target protein [disulfide, bis (2-sulfhydrylethyl] from TVL mediates the protective effects.
    CONCLUSION: Wild garlic or TVL extract has shown a protective effect on ISO-induced myocardial necrosis in rats by increasing antioxidant production confirmed with docking studies.
    Keywords:  Bis (2-sulfhydrylethyl); Disulfide; Fas-receptor; Isoproterenol; Oxidative stress; Tulbaghia violacea
    DOI:  https://doi.org/10.1016/j.jep.2019.03.049
  71. Environ Sci Pollut Res Int. 2019 Mar 26.
    Al-Brakati AY, Fouda MS, Tharwat AM, Elmahallawy EK, Kassab RB, Abdel Moneim AE.
      In the current report, we examined the potential beneficial role of soursop fruit extract (SSFE) on liver injury induced by a single paracetamol (APAP) overdose (2000 mg/kg). Thirty-five Wistar albino rats were randomly divided into five groups as follows: control, SSFE, APAP, SSFE+APAP, and silymarin (SIL)+APAP. APAP intoxication was found to elevate alanine aminotransferase, aspartate aminotransferase, alkaline phosphatase, and total bilirubin levels. Moreover, it increased the levels of malondialdehyde, nitrites, and nitrates and depleted glutathione, superoxide dismutase, catalase, glutathione reductase, and glutathione peroxidase. APAP intoxication inactivated the nuclear factor erythroid 2-related factor 2 (Nrf2) defense pathway and upregulated the expression of heme oxygenase-1 (HO-1). APAP administration enhanced the activation of nuclear factor-kappa B (NF-κB), the elevation of tumor necrosis factor-alpha and interleukin 1-beta levels, and the upregulation of inducible nitric oxide synthase mRNA expression. In addition, APAP activated the overexpression of Bax protein, increased release of cytochrome c, and the downregulation of Bcl-2 protein. Finally, APAP-induced overexpression of transforming growth factor-beta (TGF-β) further suggested enhanced liver damage. On the other hand, SSFE pretreatment attenuated these biochemical, molecular, and histopathological alterations in the liver, which might be partially due to the regulation of hepatic Nrf2/HO-1 and downregulation of NF-κB and TGF-β.
    Keywords:  Apoptosis; Fibrosis; Inflammation; Liver; Oxidative stress; Paracetamol exposure; Soursop
    DOI:  https://doi.org/10.1007/s11356-019-04935-3