bims-aporos Biomed news
on Apoptosis and reactive oxygen species
Issue of 2019‒01‒20
fifty-seven papers selected by
Gavin McStay
Staffordshire University


  1. Cytotechnology. 2019 Jan 18.
    Lee JS, Lee HA, Han JS.
      This study investigated the protective effects of Sargassum sagamianum extract (SSE) on INS-1 pancreatic β cells against high glucose-induced oxidative stress and apoptosis. Treatment with glucose at high concentrations (30 mM) caused β cell apoptosis, whereas treatment with SSE protected the β cells from high glucose-induced damage, by recovering the cell viability. Treatment with SSE at concentrations of 10-100 μg/mL decreased lipid peroxidation and intracellular reactive oxygen species and nitric oxide levels, and increased cell viability and insulin secretion in high glucose pretreated INS-1 cells in a dose-dependent manner. Moreover, SSE treatment significantly reduced the expression of pro-apoptotic Bax, cytochrome c, caspase-3, and caspase-9, while the expression of anti-apoptotic Bcl-2 increased. The type of cell death was examined by annexin V/propidium iodide staining, which revealed that SSE treatment markedly reduced high glucose-induced apoptosis. These findings suggest that SSE could be useful as a functional food, protecting pancreatic β cells against high glucose-induced oxidative stress and apoptosis.
    Keywords:  Apoptosis; INS-1; Pancreatic β cells; Sargassum sagamianum
    DOI:  https://doi.org/10.1007/s10616-019-00295-5
  2. Exp Biol Med (Maywood). 2019 Jan 12. 1535370218823549
    He J, Zhang X, Lian C, Wu J, Fang Y, Ye X.
      IMPACT STATEMENT: Nrf2 is an essential part of the defense mechanism of vertebrates and protects them from surrounding stress via participation in stimulated expression of detoxification as well as antioxidant enzymes. It also exerts a role in defending hosts from different stress in the environment, including reactive oxygen species. Our study investigates the role of exendin-4 on Nrf2 pathway as well as cell death in pancreatic β-cell and in non-obese diabetic mice. Result of study indicates exendin-4 mediates activation of Keap1-Nrf2-ARE pathway and may serve as a potential agent to treat type I diabetes mellitus. In our research, we observed excessive reactive oxygen species production, low level of cell death, and PKC phosphorylation on exendine-4 treatment. Nrf2 knockdown led to suppression of reactive oxygen species generation as well as increasing apoptosis. Moreover, siRNA-mediated Nrf2 down-regulation attenuated the suppressive effect of exendin-4 in pancreatic β-cell viability, via modulating apoptosis promoting- and counteracting-proteins, Bax, and Bcl-2.
    Keywords:  IDM; Keap1-Nrf2-ARE; PKC; ROS; apoptosis; exendin-4
    DOI:  https://doi.org/10.1177/1535370218823549
  3. Oncol Lett. 2019 Jan;17(1): 303-313
    Fu C, Li J, Aipire A, Xia L, Yang Y, Chen Q, Lv J, Wang X, Li J.
      Cistanche tubulosa has various biological functions. In the present study, the antitumor effect of water-soluble phenylethanoid glycosides of C. tubulosa (CTPG-W) on esophageal cancer was investigated. Eca-109 cells were treated with CTPG-W and the cell viability was measured by MTT assay. The apoptosis, cell cycle, mitochondrial membrane potential (Δψm) and reactive oxygen species were analyzed by flow cytometry. The levels of proteins in apoptotic pathways were detected by western blot analysis. It was determined that CTPG-W significantly reduced the viability of Eca-109 cells through the induction of apoptosis and cell cycle arrest. Following CTPG-W treatment, the Δψm of Eca-109 was notably decreased, which is associated with the upregulated levels of B-cell lymphoma-2 (Bcl-2)-associated X and downregulated levels of Bcl-2. Consequently, the levels of cytochrome c and c-Jun NH2-terminal kinase were increased, which upregulated the levels of cleaved-poly (ADP-ribose) polymerase and cleaved-caspase-3, -7 and -9, but not caspase-8. Correspondingly, the levels of reactive oxygen species in Eca-109 cells demonstrated notable changes. These results indicated that CTPG-W induced apoptosis of Eca-109 cells through a mitochondrial-dependent pathway.
    Keywords:  Cistanche tubulosa; apoptosis; cell cycle; mitochondrial-dependent pathway; phenylethanoid glycosides
    DOI:  https://doi.org/10.3892/ol.2018.9635
  4. Int J Mol Sci. 2019 Jan 11. pii: E267. [Epub ahead of print]20(2):
    Peerapanyasut W, Kobroob A, Palee S, Chattipakorn N, Wongmekiat O.
      Mitochondrial impairment ensuing from oxidative imbalance is related to adverse consequences of bisphenol A (BPA), a globally utilized industrial chemical. Recent evidence reveals sirtuin 3 (SIRT3) as a key regulator of mitochondrial homeostasis; however, its role in BPA toxicity remains unidentified. This study explored the potential benefits of N-acetylcysteine (NAC), an effective antioxidant, against BPA toxicity in the kidney and liver, and examined whether SIRT3 was involved in this condition. Male Wistar rats were fed with vehicle, BPA (5, 50 mg/kg), BPA (50 mg/kg) plus NAC (100 mg/kg) and were evaluated after 5 weeks. NAC treatment significantly diminished BPA-induced kidney and liver functional disorders, histopathological alterations, oxidative stress, and apoptosis. The increased mitochondrial reactive oxygen species, the disrupted membrane potential, the swelling, and the impaired mitochondrial fission caused by BPA were also mitigated upon concurrent treatment with NAC. The benefits of NAC were associated with enhanced AMPK-PGC-1α-SIRT3 signaling protein expressions, which led to decreased acetylation of superoxide dismutase 2 (SOD2) and increased expression of mitochondrial antioxidant manganese superoxide dismutase (MnSOD). The findings demonstrate the efficacy of NAC in protecting BPA-induced kidney and liver injury, which, in part, is mediated by activating SIRT3 and improving mitochondrial function, dynamics, and oxidative imbalance.
    Keywords:  N-acetylcysteine; bisphenol A; mitochondria; oxidative stress; sirtuin 3; toxicity
    DOI:  https://doi.org/10.3390/ijms20020267
  5. Oncogenesis. 2019 Jan 15. 8(2): 7
    Chakraborty S, Balan M, Flynn E, Zurakowski D, Choueiri TK, Pal S.
      Any imbalance between reactive oxygen species (ROS) generation and the anti-oxidant capacity lead to cellular oxidative stress. Many chemotherapeutic agents mediate their cytotoxic functions through the generation of ROS. c-Met, a receptor tyrosine kinase, is over-expressed in renal cancer and plays very crucial role(s) in its growth and survival. Here, we show that c-Met activation protected renal cancer cells from ROS, oxidative stress and cytotoxicity induced by the anti-cancer agent sorafenib (used for renal cancer treatment); and it markedly attenuated sorafenib-induced DNA damage. Activated c-Met promoted the anti-apoptotic proteins (Bcl-2 and Bcl-xL) and inhibited apoptotic cleaved caspase-3. We found that the cytoprotective function of c-Met against sorafenib-induced ROS generation and apoptosis was mediated primarily through the activation of anti-oxidant Nrf2-HO-1. c-Met promoted the nuclear localization of Nrf2 and hindered its binding with the inhibitory protein Keap1. Silencing of Nrf2 attenuated the protective action of c-Met against sorafenib-induced oxidative stress. To evaluate the physiological significance of our findings, in a tumor xenograft model, we observed that a combination treatment with pharmacological inhibitors of c-Met and it's anti-oxidant downstream effecter HO-1 markedly reduced the growth of renal tumor in vivo; it increased the oxidative stress, DNA damage and apoptotic markers in the tumor xenografts, along with reduced tumor vessel density. Our observations indicate that the c-Met-Nrf2-HO-1 pathway plays a vital role in relieving ROS-mediated oxidative stress of renal tumors. Targeting this pathway can significantly increase the oxidative stress to promote apoptotic death of cancer cells.
    DOI:  https://doi.org/10.1038/s41389-018-0116-9
  6. Int J Nanomedicine. 2019 ;14 257-270
    Behzadi E, Sarsharzadeh R, Nouri M, Attar F, Akhtari K, Shahpasand K, Falahati M.
      Background: Recently, nanomaterials have moved into biological and medicinal implementations like cancer therapy. Therefore, before clinical trials, their binding to plasma proteins like human serum albumin (HSA) and their cytotoxic effects against normal and cancer cell lines should be addressed.Methods: Herein, the interaction of magnesium oxide nanoparticles (MgO NPs) with HSA was studied by means of fluorescence spectroscopy, circular dichroism (CD) spectroscopy, and docking studies. Afterwards, the cytotoxic impacts of MgO NPs on human leukemia cell line (K562) and peripheral blood mononucleated cells (PBMCs) were evaluated by MTT and flow cytometry assays to quantify reactive oxygen species (ROS) generation and apoptosis.
    Results: It was demonstrated that MgO NPs spontaneously form a static complex with HSA molecules through hydrophobic interactions. Docking study based on the size of NPs demonstrated that different linkages can be established between MgO NPs and HSA. The CD investigation explored that MgO NPs did not alter the secondary structure of HSA. Cellular studies revealed that MgO NPs induced cytotoxicity against K562 cell lines, whereas no adverse effects were detected on PBMCs up to optimum applied concentration of MgO NPs. It was exhibited that ROS production mediated by IC50 concentrations of MgO NPs caused apoptosis-associated cell death. The pre-incubation of K562 with ROS scavenger (curcumin) inhibited the impact of MgO NPs -based apoptosis on cell fate, revealing the upstream effect of ROS in our system.
    Conclusion: In summary, MgO NPs may exhibit strong plasma distribution and mediate apoptosis by ROS induction in the cancer cell lines. These data demonstrate a safe aspect of MgO NPs on the proteins and normal cells and their application as a distinctive therapeutic approach in the cancer treatment.
    Keywords:  K562 cells; albumin; anticancer; magnesium oxide; nanoparticles; spectroscopy
    DOI:  https://doi.org/10.2147/IJN.S186428
  7. Mol Reprod Dev. 2019 Jan 13.
    Zhang T, Zhao X, Hai R, Li R, Zhang W, Zhang J.
      The low efficiency of in vitro embryo production is associated with oxidative stress induced by suboptimal culture conditions. p66Shc is a 66-kDa protein of the ShcA (Src homologous-collagen homolog) adaptor protein family, which is involved in signaling pathways involved in oxidative stress regulation, apoptosis induction, and aging. However, the functional role of p66Shc during the preimplantation development of sheep embryos is not understood. Our results showed that early-cleavage (≤28 hr) embryos had a higher developmental potential than late-cleavage (>28 hr) embryos. The poor quality of these late-cleavage embryos was associated with increased the transcripts and protein of p66Shc and decreased mitochondrial activity. In addition, exogenous hydrogen peroxide-induced oxidative stress significantly increased p66Shc protein abundance and suppressed embryonic development, which was ameliorated by antioxidant treatment. Notably, oxidative stress induced the nuclear localization of p66Shc and phosphorylated (Ser-36) p66Shc. Collectively, these observations suggest that p66Shc may be playing an important role in the regulation of oxidative stress during the preimplantation development of sheep embryos.
    Keywords:  mitochondria; p66Shc; reactive oxygen species; redox; sheep embryo
    DOI:  https://doi.org/10.1002/mrd.23110
  8. Eur J Pharmacol. 2019 Jan 15. pii: S0014-2999(19)30023-8. [Epub ahead of print]
    Xiang Y, Fan X, Zhao M, Guo Q, Guo S.
      Accumulating evidence has shown that casein kinase 2 interacting protein-1 (CKIP-1) is a pivotal regulator of apoptosis and oxidative stress. However, whether CKIP-1 is involved in regulating neuronal injury during the progression of cerebral ischemia/reperfusion injury remains unknown. In the present study, we aimed to investigate the potential role and underlying mechanism of CKIP-1 in regulating neuronal apoptosis and oxidative stress induced by oxygen-glucose deprivation/reoxygenation (OGD/R) treatment in vitro. Herein, we found that OGD/R treatment resulted in a significant increase in CKIP-1 expression in cultured hippocampal neurons. The silencing of CKIP-1 exacerbated OGD/R-induced neuronal apoptosis and production of reactive oxygen species. By contrast, CKIP-1 overexpression reduced the apoptosis and reactive oxygen species production induced by the OGD/R treatment. Mechanistically, CKIP-1 inhibited the expression of Kelch-like ECH-associated protein 1 (Keap1) and promoted the expression of nuclear factor E2-related factor 2 (Nrf2). In addition, CKIP-1 increased the activation of antioxidant response element and the expression of downstream antioxidant genes. However, Keap1 overexpression or Nrf2 knockdown partially reversed the neuroprotective effect of CKIP-1 overexpression. Taken together, our results demonstrate that CKIP-1 overexpression alleviates OGD/R-induced neuronal injury by enhancing the Nrf2-mediated anti-oxidative stress signaling pathway, revealing a neuroprotective role of CKIP-1. Our study suggests CKIP-1 as a potential therapeutic target for neuroprotection.
    Keywords:  CKIP-1; Keap1; Nrf2; oxygen-glucose deprivation/reoxygenation
    DOI:  https://doi.org/10.1016/j.ejphar.2019.01.015
  9. Oxid Med Cell Longev. 2018 ;2018 5123147
    Rehman ZU, Meng C, Sun Y, Safdar A, Pasha RH, Munir M, Ding C.
      Reactive species (RS), generally known as reactive oxygen species (ROS) and reactive nitrogen species (RNS), are produced during regular metabolism in the host and are required for many cellular processes such as cytokine transcription, immunomodulation, ion transport, and apoptosis. Intriguingly, both RNS and ROS are commonly triggered by the pathogenic viruses and are famous for their dual roles in the clearance of viruses and pathological implications. Uncontrolled production of reactive species results in oxidative stress and causes damage in proteins, lipids, DNA, and cellular structures. In this review, we describe the production of RS, their detoxification by a cellular antioxidant system, and how these RS damage the proteins, lipids, and DNA. Given the widespread importance of RS in avian viral diseases, oxidative stress pathways are of utmost importance for targeted therapeutics. Therefore, a special focus is provided on avian virus-mediated oxidative stresses. Finally, future research perspectives are discussed on the exploitation of these pathways to treat viral diseases of poultry.
    DOI:  https://doi.org/10.1155/2018/5123147
  10. Reprod Biomed Online. 2019 Jan 08. pii: S1472-6483(19)30008-2. [Epub ahead of print]
    Hezavehei M, Kouchesfahani HM, Shahverdi A, Sharafi M, Salekdeh GH, Eftekhari-Yazdi P.
      RESEARCH QUESTION: Can sublethal stress induced by nitric oxide on fresh human spermatozoa protect the functional properties of post-thaw human spermatozoa?DESIGN: Semen samples were obtained from 46 donors. Twenty semen samples were used to determine toxicity level of nitric oxide by incubation of semen with different concentrations of nitric oxide (0.01 to 400 μM). Then, 26 semen samples were cryopreserved with optimized ranges of nitric oxide: control (NO-0.00), 0.01 μM nitric oxide (NO-0.01), 0.1 μM nitric oxide (NO-0.1), 1 μM nitric oxide (NO-1), 10 μM nitric oxide (NO-10), 100 μM nitric oxide (NO-100). Frozen-thawed spermatozoa were assessed for motion characteristics, viability, morphology, apoptosis-like changes, caspase 3 activity, DNA fragmentation and intracellular reactive oxygen species levels. Fertilization potential was investigated by heterologous piezo-intracytoplasmic sperm injection (piezo-ICSI) of human spermatozoa into mouse oocytes.
    RESULTS: In fresh spermatozoa, nitric oxide did not induce a negative effect, except a significant reduction in motility and viability at 200 µM and 400 µM (P < 0.05). Cryopreservation significantly reduced sperm motility and increased reactive oxygen species, apoptosis-like changes, caspase 3 activity, and DNA damage (P < 0.05). NO-0.01 significantly increased total and progressive motility versus the other groups (P < 0.05). The lowest percentage of caspase 3 activity was in the NO-0.01 and NO-0.1 compared with the other freezing groups. In the fertilization trial, the rate of two-cell embryo formation after heterologous piezo-ICSI was higher (P < 0.05) in NO-0.01 (69%) versus controls (42%).
    CONCLUSIONS: Sublethal oxidative stress induced by nitric oxide might improve human sperm function after cryopreservation.
    Keywords:  Apoptosis; Freezing; Motility; Nitric oxide; Sublethal stress
    DOI:  https://doi.org/10.1016/j.rbmo.2018.11.029
  11. Oncol Lett. 2019 Jan;17(1): 757-764
    Qiu M, Zhang S, Ke L, Tang H, Zeng X, Liu J.
      The aim of the present study was to investigate the influence of the nitric oxide donor prodrug JS-K (C13H16N6O8) on Taxol-induced apoptosis in prostate cancer cells, and to investigate a potential reactive oxygen species (ROS)-associated mechanism. The effect of JS-K on the anticancer activity of Taxol was assessed in prostate cancer cells; cell viability, colony formation, apoptosis, ROS generation and expression levels of apoptosis-associated proteins were investigated. The function of ROS accumulation in the combined effects of JS-K and Taxol was determined using the antioxidant N-acetylcysteine (NAC) and the pro-oxidant oxidized glutathione (GSSG). The results of the present study demonstrated that JS-K was able to increase Taxol-induced suppression of prostate cancer cell proliferation, apoptosis, ROS accumulation and upregulation of apoptosis-associated proteins. Furthermore, NAC reversed the effect of JS-K on Taxol-induced apoptosis and conversely, the pro-oxidant GSSG exacerbated the effect of JS-K on Taxol-induced apoptosis in prostate cancer cells. In conclusion, JS-K enhances the chemosensitivity of prostate cancer cells to Taxol, via the upregulation of intracellular ROS.
    Keywords:  JS-K; Taxol; chemosensitivity; prostate cancer cells; reactive oxygen species
    DOI:  https://doi.org/10.3892/ol.2018.9684
  12. Exp Ther Med. 2019 Jan;17(1): 663-670
    Guo Z, Yuan Y, Guo Y, Wang H, Song C, Huang M.
      Nischarin (NISCH) is a cytoplasmic protein known to serve an inhibitory role in breast cancer cell apoptosis, migration and invasion. Recently, NISCH has been reported to be involved in the regulation of spinal cord injury (SCI). However, the molecular mechanism is still unclear. Oxidative stress contributes to tissue injury and cell apoptosis during the development of various diseases, including SCI. The aim of the present study was to investigate the role of NISCH in the regulation of apoptosis induced by oxidative stress in PC12 cells. H2O2 was used to establish an oxidative stress model in PC12 cells. Apoptosis levels were examined using flow cytometry analysis, and the expression of NISCH, Bcl-2, Bcl-2-associated X (Bax) and caspase-3 were examined using western blot and immunofluorescence staining analyses. The results demonstrated that treatment with 100 µM H2O2 significantly increased the apoptotic rate and expression of NISCH in PC12 cells. At 48 h following incubation with 100 µM H2O2, NISCH downregulation partially inhibited apoptosis of PC12 cells. In addition, the expression of Bcl-2 was significantly reduced and the expression of Bax and caspase-3 were significantly increased by H2O2 treatment. These effects were also partially inhibited by the downregulation of NISCH. The authors of the present study therefore hypothesize that NISCH may function as a pro-apoptotic protein that participates in the regulation of oxidative stress, and NISCH downregulation may protect cells from oxidative stress-induced apoptosis.
    Keywords:  Bcl-2/Bcl-2-associated X signaling; apoptosis; hydrogen peroxide; nischarin; oxidative stress
    DOI:  https://doi.org/10.3892/etm.2018.7017
  13. Biochem Biophys Res Commun. 2019 Jan 12. pii: S0006-291X(19)30021-X. [Epub ahead of print]
    Wang Z, Li J, Wang Y, Liu Q.
      Cell cycle dysregulation is typical in human cancers, and CDK4/6 inhibitors targeting cell cycle have potential antiapoptosis effect. The aim of this work is to identify the regulatory effect of Palbociclib on apoptosis of H9c2 cells induced by high glucose (HG) and to elucidate the fundamental mechanisms. It was observed that Palbociclib decreased intracellular ROS production, augmented mitochondrial membrane potential and hindered apoptosis of H9c2 cells. Palbociclib increased the Bcl-2/Bax ratio, diminished the expressions of Bax and cleaved-caspase-3, and affected the RB phosphorylation and p53 expression. Altogether, the anti-apoptotic efficacy of Palbociclib could be attributed in part to the modulation of the mitochondria apoptotic pathway.
    Keywords:  Apoptosis; Cell cycle; High glucose; Palbociclib; ROS generation
    DOI:  https://doi.org/10.1016/j.bbrc.2019.01.015
  14. Eur Rev Med Pharmacol Sci. 2019 Jan;pii: 16786. [Epub ahead of print]23(1): 378-388
    Yang Y, Wu ZZ, Cheng YL, Lin W, Qu C.
      OBJECTIVE: Age-related macular degeneration (AMD) is mainly characterized by dysfunction of retinal pigment epithelium (RPE) cells. This study aimed to investigate the protective effects of resveratrol on oxidative damaged RPE cells.MATERIALS AND METHODS: Human D407 cells were divided into normal control (NC), H2O2 treated (H2O2, treating with H2O2 at a final concentration of 200 mol/l) and resveratrol treatment groups (treating with resveratrol at a concentration of 12.5, 25, 50 and 100 mg/l). Malondialdehyde (MDA) and superoxide dismutase (SOD) activities were examined using enzyme-linked immunosorbent assay (ELISA). 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide (MTT) and cell count kit-8 (CCK-8) were used to examine cell viability. Cell cycle phase distribution and apoptosis of D407 cells were evaluated using flow cytometry assay. B-cell lymphoma-2 (Bcl-2) and cleaved caspase 3 expression were detected using quantitative real-time PCR (qRT-PCR) and Western blot assay, respectively.
    RESULTS: Resveratrol significantly decreased inhibitive ratios of D407 cell growth compared to that of H2O2 group (p<0.05). Resveratrol significantly increased SOD activity compared to that of H2O2 group (p<0.05). Resveratrol significantly reduced MDA activity compared to that of H2O2 group (p<0.05). Resveratrol affected cell cycle phase distribution of D407 cells compared to that of H2O2 group (p<0.05). Resveratrol significantly decreased the early stage and late stage apoptosis rates compared to that of H2O2 group (p<0.05). Resveratrol significantly enhanced Bcl-2 levels and decreased cleaved caspase 3 levels compared to that of H2O2 group (p<0.05).
    CONCLUSIONS: Resveratrol protected against the oxidative damage of RPE cells by modulating SOD/MDA activity and activating Bcl-2 expression.
    DOI:  https://doi.org/10.26355/eurrev_201901_16786
  15. J Virol. 2019 Jan 16. pii: JVI.01784-18. [Epub ahead of print]
    Zhang Y, Sun R, Geng S, Shan Y, Li X, Fang W.
      Our previous studies have demonstrated that porcine circovirus type 2 (PCV2) triggers unfolded protein response (UPR) in PK-15 cells by activating the PERK/eIF2α pathway of endoplasmic reticulum stress (ER stress) that, in turn, facilitates viral replication. PCV2 is found to cause oxidative stress and up-regulation of cytoplasmic Ca2+ The virus is reported to employ its ORF3 to induce apoptosis. We wonder if and how PCV2-induced UPR would lead to apoptosis independent of ORF3. Using an ORF3-deficient PCV2 mutant (ΔORF3), apoptotic responses in infected PK-15 and PAM cells were still apparent, though lower than its parental PCV2 strain. We hypothesized that apoptosis induced by ΔORF3 might result from UPR. We found that ΔORF3-induced apoptosis was significantly reduced when the infected cells were treated with a selective PERK blocker GSK2606414 (GSK) or a general ER stress attenuator 4-phenylbutyrate (4-PBA). Such treatments also ameliorated elevation of cytoplasmic Ca2+ and reactive oxygen species (ROS) in PK-15 and PAM cells, two predisposing factors of apoptosis via disruption of the ER-mitochondrial units. Treatment of ΔORF3-infected cells with GSK and 4-PBA also decreased mitochondrial Ca2+ load and increased mitochondrial membrane potential (MMP). By transient expression of the structural protein capsid (Cap) in combination with PERK silencing, we found that Cap induced collapse of MMP and mitochondrial apoptosis could result from UPR and elevation of Ca2+ and ROS that were inhibitable by down-regulation of PERK. We propose that PCV2-driven ER stress is Cap-dependent and could lead to mitochondrial apoptotic response independent of ORF3 via perturbation of intracellular Ca2+ homeostasis and accumulation of ROS.IMPORTANCE Porcine circovirus type 2 (PCV2) encoded protein ORF3, a putative protein having pro-apoptotic activity. Our early studies have shown that PCV2 infection triggers ER stress via selective activation of the PERK pathway, a branch of ER stress pathways, in permissive cells for enhanced replication and its infection increased level of cytosolic Ca2+ and ROS. Here we clearly show that PCV2 infection or capsid protein (Cap) expression induces ORF3-independent apoptosis via increased cytosolic and mitochondrial Ca2+ and cellular ROS as a result of activation of PERK pathway.
    DOI:  https://doi.org/10.1128/JVI.01784-18
  16. Exp Clin Endocrinol Diabetes. 2019 Jan 14.
    Xu S, Bao W, Men X, Liu Y, Sun J, Li J, Liu H, Cai H, Zhang W, Lou J, Peng L.
      Demyelination resulting from Schwann cell injury is a main pathological feature of diabetic neuropathy, and a key contributor to this process may be inflammation due to advanced glycation end products (AGEs). Therefore, protection by anti-inflammation agents is anticipated. In this study, we showed that interleukin-10 (IL-10), an anti-inflammatory cytokine, inhibits apoptosis of Schwann cells induced by AGEs in vitro. We isolated and cultured Schwann cells from rat sciatic nerves. As detected by flow cytometry, apoptosis of Schwann cells markedly increased following incubation with AGEs for 48 h. However, pretreatment with IL-10 inhibited AGE-induced apoptosis. The effect of IL-10 on NF-κB, which is a very important regulator of inflammation, was also evaluated, and results showed high levels of phospho-NF-κB and nuclear localization of NF-κB in cells incubated with AGEs but low levels of phospho-NF-κB and cytoplasmic localization in the cells incubated with IL-10, indicating the activation of NF-κB by AGEs and inhibition of NF-κB by IL-10. Moreover, incubating Schwann cells with an NF-κB inhibitor (caffeic acid phenethyl ester) for 30 min before adding AGEs mimicked IL-10, lowering the amount of reactive oxygen species and activity of caspase-3 and also decreasing apoptosis in Schwann cells. These results indicate that IL-10 may protect Schwann cells against AGE-induced apoptosis by attenuating oxidative stress via the inhibition of activation of NF-κB.
    DOI:  https://doi.org/10.1055/a-0826-4374
  17. Free Radic Biol Med. 2019 Jan 10. pii: S0891-5849(18)31303-0. [Epub ahead of print]
    Kikuchi R, Iwai Y, Tsuji T, Watanabe Y, Koyama N, Yamaguchi K, Nakamura H, Aoshiba K.
      The tumor microenvironment has previously been reported to be hypercapnic (as high as ~84mmHg), although its effect on tumor cell behaviors is unknown. In this study, high CO2 levels, ranging from 5% to 15%, protected lung cancer cells from anticancer agents, such as cisplatin, carboplatin and etoposide, by suppressing apoptosis. The cytoprotective effect of a high CO2 level was independent of acidosis and was due to mitochondrial metabolic reprogramming that reduced mitochondrial respiration, as assessed by oxygen consumption, oxidative phosphorylation, mitochondrial membrane and oxidative potentials, eventually leading to reduced reactive oxidant species production. In contrast, high CO2 levels did not affect cisplatin-mediated DNA damage responses or the expression of Bcl-2 family proteins. Although high CO2 levels inhibited glycolysis, this inhibition was not mechanistically involved in high CO2-mediated reductions in mitochondrial respiration, because a high CO2 concentration inhibited isolated mitochondria. A cytoprotective effect of high CO2 levels on mitochondria DNA-depleted cells was not noted, lending support to our conclusion that high CO2 levels act on mitochondria to reduce the cytotoxicity of anticancer agents. High CO2-mediated cytoprotection was also noted in a 3D culture system. In conclusion, the hypercapnic tumor microenvironment reprograms mitochondrial respiratory metabolism causing chemoresistance in lung cancer cells. Thus, tumor hypercapnia may represent a novel target to improve chemosensitivity.
    Keywords:  CO(2); Carboplatin; Cisplatin; Reactive oxygen species
    DOI:  https://doi.org/10.1016/j.freeradbiomed.2019.01.014
  18. Oxid Med Cell Longev. 2018 ;2018 6121328
    Yuan YG, Zhang S, Hwang JY, Kong IK.
      Silver nanoparticles (AgNPs) are widely used metal nanoparticles in health care industries, particularly due to its unique physical, chemical, optical, and biological properties. It is used as an antibacterial, antiviral, antifungal, and anticancer agent. Camptothecin (CPT) and its derivatives function as inhibitors of topoisomerase and as potent anticancer agents against a variety of cancers. Nevertheless, the combined actions of CPT and AgNPs in apoptosis in human cervical cancer cells (HeLa) have not been elucidated. Hence, we investigated the synergistic combinatorial effect of CPT and AgNPs in human cervical cancer cells. We synthesized AgNPs using sinigrin as a reducing and stabilizing agent. The synthesized AgNPs were characterized using various analytical techniques. The anticancer effects of a combined treatment with CPT and AgNPs were evaluated using a series of cellular and biochemical assays. The expression of pro- and antiapoptotic genes was measured using real-time reverse transcription polymerase chain reaction. The findings from this study revealed that the combination of CPT and AgNPs treatment significantly inhibited cell viability and proliferation of HeLa cells. Moreover, the combination effect significantly increases the levels of oxidative stress markers and decreases antioxidative stress markers compared to single treatment. Further, the combined treatment upregulate various proapoptotic gene expression and downregulate antiapoptotic gene expression. Interestingly, the combined treatment modulates various cellular signaling molecules involved in cell survival, cytotoxicity, and apoptosis. Overall, these results suggest that CPT and AgNPs cause cell death by inducing the mitochondrial membrane permeability change and activation of caspase 9, 6, and 3. The synergistic cytotoxicity and apoptosis effect seems to be associated with increased ROS formation and depletion of antioxidant. Certainly, a combination of CPT and AgNPs could provide a beneficial effect in the treatment of cervical cancer compared with monotherapy.
    DOI:  https://doi.org/10.1155/2018/6121328
  19. PLoS One. 2019 ;14(1): e0210274
    Chang TC, Wei PL, Makondi PT, Chen WT, Huang CY, Chang YJ.
      Advanced colorectal cancer (CRC) survival rates are still low despite advances in cytotoxic and targeted therapies. The development of new effective or alternative therapies is therefore urgently needed. Bromelain, an extract of pineapple, was shown to have anticancer effects, but its mechanisms in CRC have not been fully explored. Therefore, the roles of bromelain in CRC progression were investigated using different CRC cell lines, a zebrafish model, and a xenograft mouse model. The anticancer mechanisms were explored by assessing the role of bromelain in inducing reactive oxygen species (ROS), superoxide, autophagosomes, and lysosomes. The role of bromelain in the induction of apoptosis was also assessed. It was found that bromelain inhibited CRC cell growth in cell lines and tumor growth in the zebrafish and xenograft mouse models. It also induced high levels of ROS and superoxide, plus autophagosome and lysosome formation. High levels of apoptosis were also induced, which were associated with elevated amounts of apoptotic proteins like apoptotic induction factor, Endo G, and caspases-3, -8, and -9 according to a qPCR analysis. In a Western blot analysis, increases in levels of ATG5/12, beclin, p62, and LC3 conversion rates were found after bromelain treatment. Levels of cleaved caspase-3, caspase-8, caspase-9, and poly(ADP ribose) polymerase (PARP)-1 increased after bromelain exposure. This study explored the role of bromelain in CRC while giving insights into its mechanisms of action. This compound can offer a cheap alternative to current therapies.
    DOI:  https://doi.org/10.1371/journal.pone.0210274
  20. Curr Pharm Des. 2019 Jan 12.
    Coppolino G, Leonardi G, Andreucci M, Bolignano D.
      BACKGROUND: ROS are highly reactive and cause in single cells: protein alteration, DNA damage, cellular senescence and apoptosis; while the effect of ROS in biological tissues leads to a harmful oxidation effect on all their biochemical components: lipids, proteins, carbohydrates, and nucleic acids. Oxidative stress plays a role in the pathophysiology of renal impairment and is a mediator of CKD progression; furthermore during substitutive therapy with haemodialysis or peritoneal dialysis and in case of transplantation, organism continues to be exposed to oxidation causing the development of major systemic comorbidities in particular cardiovascular diseases.METHODS: In this review we summarized some information regarding the link between kidney and oxidative stress. The kidneys maintain plasma homeostasis by filtering plasma and reabsorbing approximately 99% of the filtrate back into the plasma. This enormous reabsorption work requires a great consume of molecular oxygen (O2) due to active transport.
    CONCLUSION: Accruing evidences indicated the kidney as a fundamental organ in reactive oxygen species (ROS) production.
    Keywords:  Chronic Kidney Disease (CKD) ; Oxidative stress; renal function
    DOI:  https://doi.org/10.2174/1381612825666190112165206
  21. Nanomaterials (Basel). 2019 Jan 11. pii: E91. [Epub ahead of print]9(1):
    Zhang C, Wang Y, Zhao Y, Liu H, Zhao Y, Li X, Lin Q.
      The chemotherapy of stimuli-responsive drug delivery systems (SDDSs) is a promising method to enhance cancer treatment effects. However, the low efficiency of chemotherapy drugs and poor degradation partly limit the application of SDDSs. Herein, we report doxorubicin (DOX)-loading mixed micelles for biotin-targeting drug delivery and enhanced photothermal/photodynamic therapy (PTT/PDT). Glutathione (GSH)-responsive mixed micelles were prepared by a dialysis method, proportionally mixing polycaprolactone-disulfide bond-biodegradable photoluminescent polymer (PCL-SS-BPLP) and biotin-polyethylene glycol-cypate (biotin-PEG-cypate). Chemically linking cypate into the mixed micelles greatly improved cypate solubility and PTT/PDT effect. The micelles also exhibited good monodispersity and stability in cell medium (~119.7 nm), low critical micelles concentration, good biodegradation, and photodecomposition. The high concentration of GSH in cancer cells and near-infrared light (NIR)-mediated cypate decomposition were able to achieve DOX centralized release. Meanwhile, the DOX-based chemotherapy combined with cypate-based NIR-triggered hyperthermia and reactive oxygen species could synergistically induce HepG2 cell death and apoptosis. The in vivo experiments confirmed that the micelles generated hyperthermia and achieved a desirable therapeutic effect. Therefore, the designed biodegradable micelles are promising safe nanovehicles for antitumor drug delivery and chemo/PTT/PDT combination therapy.
    Keywords:  biodegradable; cypate; photodynamic therapy; photothermal therapy; stimuli-responsive drug delivery systems
    DOI:  https://doi.org/10.3390/nano9010091
  22. Int J Mol Sci. 2019 Jan 14. pii: E318. [Epub ahead of print]20(2):
    Kang SJ, Rhee WJ.
      Autophagy is a natural physiological process, and it induces the lysosomal degradation of intracellular components in response to environmental stresses, including nutrient starvation. Although an adequate autophagy level helps in cell survival, excessive autophagy triggered by stress such as starvation leads to autophagy-mediated apoptosis. Chinese hamster ovary (CHO) cells are widely used for producing biopharmaceuticals, including monoclonal antibodies. However, apoptosis induced by high stress levels, including nutrient deficiency, is a major problem in cell cultures grown in bioreactors, which should be overcome. Therefore, it is necessary to develop a method for suppressing excessive autophagy and for maintaining an appropriate autophagy level in cells. Therefore, we investigated the effect of silkworm storage protein 1 (SP1), an antiapoptotic protein, on autophagy-mediated apoptosis. SP1-expressing CHO cells were generated to assess the effect and molecular mechanism of SP1 in suppressing autophagy. These cells were cultured under starvation conditions by treatment with Earle's balanced salt solution (EBSS) to induce autophagy. We observed that SP1 significantly inhibited autophagy-mediated apoptosis by suppressing caspase-3 activation and reactive oxygen species generation. In addition, SP1 suppressed EBSS-induced conversion of LC3-I to LC3-II and the expression of autophagy-related protein 7. Notably, basal Beclin-1 level was significantly low in the SP1-expressing cells, indicating that SP1 regulated upstream events in the autophagy pathway. Together, these findings suggest that SP1 offers a new strategy for overcoming severe autophagy-mediated apoptosis in mammalian cells, and it can be used widely in biopharmaceutical production.
    Keywords:  apoptosis; autophagy; silkworm; storage protein 1
    DOI:  https://doi.org/10.3390/ijms20020318
  23. Int J Reprod Biomed (Yazd). 2018 Sep;16(9): 577-586
    Kaushik K, Kaushal N, Kalla NR.
      Background: Increased scrotal temperature can disrupt spermatogenesis leading to male infertility. Germ cells being heat sensitive maintain their genetic integrity via protective mechanism originated from the cell itself or by means of cell death. However, qualitative differentiation of how reactive oxygen species and antioxidant enzymes regulate signaling pathways of cellular damage including DNA fragmentation at varied temperatures remains unexplored.Objective: The study was designed to evaluate the effects of heat mediated oxidative stress on male germ cells. Also, the time-dependent qualitative variation in the germ cell death was studied.
    Materials and Methods: Thirty male Wistar rats were randomly segregated into five major groups (n=6/each) i.e., Control, 30, 45, 60, and 90-min counterparts according to heat treatment protocol. Quantitation of DNA and DNA ladder studies was performed along with various biochemical parameter like lipid peroxidation (LPO), glutathione, catalase, superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), glutatione reductase (GR), glutathione-s-transferase (GST).
    Results: Animals receiving heat treatment for 30-min and 45-min revealed systematic and gradual response to heat stress; whereas, 60-min and 90-min treated animals showed a typical and abrupt change of the internal milieu of germ cells. Laddering and smearing effect of damaged DNA in 30 and 45 min and 60 and 90 min heat treated animals was seen respectively.
    Conclusion: As the duration of heat treatment increases, the rate of apoptosis reaches an optimum level, and a further increase in the duration of heat treatment converted the mode of cell death from apoptosis to necrosis, implicitly due to severe oxidative attack.
    Keywords:  Antioxidant enzymes; Apoptosis; Heat stress; Necrosis; Testes
  24. Med Sci Monit. 2019 Jan 15. 25 453-459
    Shan S, Tian L, Fang R.
      BACKGROUND Chlorogenic acid (CGA), a dietary polyphenol derived from many plants, has been previously reported to exert neuroprotective properties. However, its pharmacological role in Parkinson's disease (PD) and the underlying mechanisms are unclear. MATERIAL AND METHODS In the present study, we investigated the beneficial effects of CGA against the toxicity of 6-hydroxydopamine (6-OHDA) in animal and cellular models. One week after 6-OHDA administration, the behavioral activities of rats were determined by rotarod test and apomorphine-induced rotational test. The viability and apoptosis of SH-SY5Y cells following 6-OHDA exposure were determined by MTT assay and annexin V-FITC/PI double staining, respectively. The activities of antioxidant enzymes in the rat striatal tissues and SH-SY5Y cells were detected by ELISA. RESULTS The results demonstrated that 6-OHDA-induced PD-like behavioral impairments of rats were significantly forestalled by CGA administration. The increased apoptosis and reduced activities of antioxidant enzymes in the striatum of 6-OHDA-lesioned rats were also attenuated by CGA. Moreover, in an in vitro experiment, the impaired viability and enhanced apoptosis of 6-OHDA-injured SH-SY5Y cells were significantly restored by CGA pretreatment. In addition, CGA also obstructed 6-OHDA-induced ROS production and endoplasmic reticulum (ER) stress in SH-SY5Y cells. CONCLUSIONS Taken together, these data show that CGA might be an effective neuroprotective compound that mitigates oxidative stress and ER stress in PD.
    DOI:  https://doi.org/10.12659/MSM.911166
  25. Anticancer Drugs. 2019 Jan 11.
    Liese J, Hinrichs TM, Lange M, Fulda S.
      Hepatocellular carcinoma (HCC) is the most common liver malignancy, and the lack of effective chemotherapies underlines the need for novel therapeutic approaches for this disease. Recently, we discovered a novel synergistic induction of cell death by combining sorafenib, the only routinely used palliative chemotherapeutic agent, and the triterpenoid oleanolic acid (OA). However, the underlying mechanisms of action have remained obscure. Here, we report that sorafenib and OA acted in concert to trigger mitochondria-mediated apoptotic cell death, which is dependent on reactive oxygen species (ROS). Sorafenib/OA cotreatment significantly increased ROS production, which was prevented by the ROS scavengers α-tocopherol and MnTBAP. Importantly, rescue experiments showed that ROS were required for sorafenib/OA-induced apoptosis as ROS scavengers protected HCC cells against cell death. In addition, sorafenib and OA cotreatment cooperated to decrease myeloid cell leukaemia-1 expression and to activate Bak, two events that were prevented by ROS scavengers. Bak activation was accompanied by the loss of mitochondrial membrane potential, followed by PARP cleavage, DNA fragmentation and, finally, apoptotic cell death in HCC cells. By providing new insights into the molecular regulation of sorafenib/OA-mediated and ROS-dependent cell death, our study contributes toward the development of novel treatment strategies to overcome sorafenib resistance in HCC.
    DOI:  https://doi.org/10.1097/CAD.0000000000000750
  26. J Mol Neurosci. 2019 Jan 15.
    Zhou B, Liu HY, Zhu BL.
      We attempted to explore the possible effects of SOCS3 (suppressor of cytokine signaling 3)-modified bone marrow mesenchymal stem cells (BMSCs) on the hypoxic injury of rat adrenal gland pheochromocytoma (PC-12) cells. PC12 cells were cultured with EGFP (enhanced green fluorescent protein)-BMSCs and SOCS3-BMSCs respectively under hypoxia in vitro and classified into control, hypoxia, EGFP-BMSCs, and SOCS3-BMSC groups. CCK-8, Hoechst 33258 staining, and Annexin V-FITC/PI staining were assessed to measure the viability and apoptosis of hypoxia-induced PC12 cells. The JAK/STAT3 pathway- and apoptosis-related proteins were identified by Western blot. Finally, rat models of permanent middle cerebral artery occlusion (pMCAO) were established to verify the potential influences of SOCS3-BMSCs in vivo. SOCS3-modified BMSCs can stably express SOCS3 protein. EGFP-BMSCs, especially SOCS3-BMSCs, can improve cell viability and SOD content, and reduce cell apoptosis, LDH viability, and MDA content in hypoxia-induced PC12 cells (all P < 0.05). Besides, EGFP-BMSCs and SOCS3-BMSCs decreased cleaved caspase-3 level and increased Bcl-2/Bax of hypoxia-induced PC12 cells, while SOCS3-BMSCs could also elevate SOCS3 protein and reduce p-STAT3 protein level in hypoxia-induced PC12 cells (all P < 0.05). In vivo experiments confirmed that EGFP-BMSCs, particularly SOCS3-BMSCs, could ameliorate infarct size and inhibit neuronal apoptosis of different degrees in pMACO rats (all P < 0.05). SOCS3-modified BMSCs can alleviate oxidative stress, improve cell viability, and reduce neuronal apoptosis by downregulation of JAK/STAT3 pathway, thereby exerting the neuroprotective role in ischemic brain injury.
    Keywords:  BMSCs; Hypoxia; JAK/STAT3; PC12 cells; SOCS3
    DOI:  https://doi.org/10.1007/s12031-018-1243-7
  27. Biochem Biophys Res Commun. 2019 Jan 08. pii: S0006-291X(18)32851-1. [Epub ahead of print]
    Jianrong S, Yanjun Z, Chen Y, Jianwen X.
      Ischemic stroke is the second most common cause of death, a major cause of acquired disability in adults. However, the pathogenesis that contributes to ischemic stroke has not been fully understood. Dual-specificity phosphatase 14 (DUSP14, also known as MKP6) is a MAP kinase phosphatase, playing important role in regulating various cellular processes, including oxidative stress and inflammation. However, its effects on cerebral ischemia/reperfusion (IR) are unclear. In the study, we found that DUSP14 expression was decreased responding to IR surgery. Over-expressing DUSP14 reduced the infarction volume of cerebral IR mice. Cognitive dysfunction was also improved in mice with DUSP14 over-expression. Promoting DUSP14 expression markedly reduced the activation of glial cells, as evidenced by the decreases in GFAP and Iba-1 expressions in mice with cerebral IR injury. In addition, inflammatory response induced by cerebral IR injury was inhibited in DUSP14 over-expressed mice, as proved by the reduced expression of tumor necrosis factor (TNF)-α and interleukin 1β (IL-1β). Furthermore, oxidative stress was markedly reduced by DUSP14 over-expression through elevating nuclear factor-erythroid 2 related factor 2 (Nrf-2) signaling pathway. Importantly, we found that DUSP14 could interact with Nrf-1, which thereby protected mice against cerebral IR injury. In vitro, we also found that repressing Nrf-2 expression abrogated DUSP14 over-expression-reduced inflammation and ROS generation. Consistent with the anti-inflammatory effect of DUSP14, reducing the production of reactive oxygen species (ROS) also down-regulated TNF-α and IL-1β expressions. Collectively, elevated DUSP14 alleviated brain damage from cerebral IR injury through Nrf-2-regulated anti-oxidant signaling pathway, and the restraining of inflammatory response. These results suggested that DUSP14 might be a potential therapeutic target to prevent ischemic stroke.
    Keywords:  Cerebral IR injury; DUSP14; Inflammation; Nrf-2; Oxidative stress
    DOI:  https://doi.org/10.1016/j.bbrc.2018.12.170
  28. Chem Biol Interact. 2019 Jan 14. pii: S0009-2797(18)31532-1. [Epub ahead of print]
    Xia X, Xiang X, Huang F, Zhang Z, Han L.
      Canolol (4-vinylsyringol), extracted form crude canola oil, is the promising drug toward cancer prevention and treatment. The current studies focus on the role of COX-2 signaling pathway in canolol-induced apoptosis in cancer cells. It is still unknown whether mitochondria and MAPK signaling pathways are involved. To elucidate the roles of above signaling pathways in canolol-induced apoptosis in cancer cells, human cervical carcinoma cell line HeLa and HeLa xenograft tumor model are adopted. Canolol induced apoptosis of HeLa cells and inhibited tumor growth with low systemic adverse effect, accompanying with excess generation of intracellular ROS and lysosome rupture. The results in vitro and in vivo confirmed that canolol-induced apoptosis involved MAPK signaling pathways mediated mitochondrial signaling pathway activation. In conclusion, these data showed that canolol induced apoptosis in HeLa cells through ROS-MAPK mediated mitochondrial signaling pathway, providing a view of the potential application of canolol as an anticancer agent.
    Keywords:  Apoptosis; Canolol; MAPK; Mitochondria; ROS
    DOI:  https://doi.org/10.1016/j.cbi.2019.01.016
  29. Exp Ther Med. 2019 Jan;17(1): 551-557
    Xie Y, Ji R, Han M.
      Myocardial infarction is a leading cause of mortality worldwide, while myocardial ischemia and timely reperfusion contribute to myocardial injury. The mitochondria are involved in the injury and mediate the apoptosis of cardiomyocytes. In order to develop novel therapeutic approaches for myocardial infarction, the present study evaluated the myocardial protective effects of eriodictyol and investigated relevant mechanisms in H9c2 cardiomyocytes. As a result, eriodictyol was observed to improve the H9c2 cardiomyocyte viability and block the leakage of cytosolic lactate dehydrogenase under hypoxia/reoxygenation. In addition, the dysfunction of mitochondria induced by hypoxia/reoxygenation was ameliorated by eriodictyol through suppressing the overload of intracellular Ca2+, preventing overproduction of reactive oxygen species, blocking mitochondrial permeability transition pore opening, increasing mitochondrial membrane potential level and decreasing ATP depletion. Finally, the apoptosis of H9c2 cardiomyocyte induced by hypoxia/reoxygenation was prevented by eriodictyol through upregulation of the expression of B-cell lymphoma-2 (Bcl-2) and downregulation of the expression levels of Bcl-2-associated X protein and caspase-3. These results provided evidence for further investigation on myocardial protection and the treatment of myocardial infarction using eriodictyol.
    Keywords:  H9c2 cardiomyocyte; eriodictyol; hypoxia/reoxygenation; mitochondrial dysfunction; myocardial protection
    DOI:  https://doi.org/10.3892/etm.2018.6918
  30. Redox Biol. 2019 Jan 03. pii: S2213-2317(18)31101-7. [Epub ahead of print]21 101095
    Yu L, Liang Q, Zhang W, Liao M, Wen M, Zhan B, Bao H, Cheng X.
      The induction of mitochondrial reactive oxygen species (mtROS) by hyperglycemia is a key event responsible for endothelial activation and injury. Heat shock protein 22 (HSP22) is a stress-inducible protein associated with cytoprotection and apoptosis inhibition. However, whether HSP22 prevents hyperglycemia-induced vascular endothelial injury remains unclear. Here, we investigated whether HSP22 protects the vascular endothelium from hyperglycemia-induced injury by reducing mtROS production. We used a high-fat diet and streptozotocin injection model to induce type 2 diabetes mellitus (T2DM, metabolic syndrome) and exposed human umbilical vein endothelial cells (HUVECs) to high glucose following overexpression or silencing of HSP22 to explore the role of HSP22. We found that HSP22 markedly inhibited endothelial cell activation and vascular lesions by inhibiting endothelial adhesion and decreasing cytokine secretion. We performed confocal microscopy and flow cytometry assays using HUVECs and showed that HSP22 attenuated mtROS and mitochondrial dysfunction in hyperglycemia-stimulated endothelial cells. Mechanistically, using the mtROS inhibitor MitoTEMPO, we demonstrated that HSP22 suppressed endothelial activation and injury by eliminating hyperglycemia-mediated increases in mtROS. Furthermore, we found that HSP22 maintained the balance of mitochondrial fusion and fission by mitigating mtROS in vitro. HSP22 attenuated the development of vascular lesions by suppressing mtROS-mediated endothelial activation in a T2DM mouse model. This study provides evidence that HSP22 may be a promising therapeutic target for vascular complications in T2DM.
    Keywords:  HSP22; Hyperglycemia; Inflammation; Oxidative stress; T2DM; mtROS
    DOI:  https://doi.org/10.1016/j.redox.2018.101095
  31. Free Radic Biol Med. 2019 Jan 09. pii: S0891-5849(18)32332-3. [Epub ahead of print]
    Florean C, Song S, Dicato M, Diederich M.
      Redox changes and generation of reactive oxygen species (ROS) are part of normal cell metabolism. While low ROS levels are implicated in cellular signaling pathways necessary for survival, higher levels play major roles in cancer development as well as cell death signaling and execution. A role for redox changes in apoptosis has been long established; however, several new modalities of regulated cell death have been brought to light, for which the importance of ROS production as well as ROS source and targets are being actively investigated. In this review, we summarize recent findings on the role of ROS and redox changes in the activation and execution of two major forms of regulated cell death, necroptosis and ferroptosis. We also discuss the potential of using modulators of these two forms of cell death to exacerbate ROS as a promising anticancer therapy.
    Keywords:  ROS; anticancer therapy; cancer; ferroptosis; necroptosis; redox changes; regulated cell death
    DOI:  https://doi.org/10.1016/j.freeradbiomed.2019.01.008
  32. Future Oncol. 2019 Jan 16.
    Wang L, Yang X, Yang C, Gao J, Zhao Y, Cheng C, Zhao G, Liu S.
      AIM: This study investigated the effect and mechanism of cold atmospheric plasma (CAP)-activated media on A431 and HaCaT cells.MATERIALS & METHODS: Phosphate-buffered saline (PBS) and Dulbecco's Modified Eagle's Medium (DMEM) were treated by CAP to get CAP-activated media. A431 and HaCaT were incubated by CAP-activated media for 2 h. MTT, Annexin-V and propidium iodide (PI), Western blot and reactive oxygen species (ROS) detection assay were utilized to demonstrate the effect.
    RESULTS: The CAP-activated media decreased the proliferation of A431 cells in a dose/time-dependent manner. And the CAP-activated media could induce apoptosis in A431 cells. CAP-activated phosphate-buffered saline could increase intracellular ROS level but not CAP-activated DMEM.
    CONCLUSION: CAP-activated media could induce apoptosis in A431 cells by production of ROS.
    Keywords:  CAP-activated media; cold atmospheric plasma; cutaneous squamous cell carcinoma; reactive oxygen species
    DOI:  https://doi.org/10.2217/fon-2018-0419
  33. J Pharmacol Sci. 2018 Dec 22. pii: S1347-8613(18)30223-8. [Epub ahead of print]
    Jiang P, Chen L, Sun J, Li J, Xu J, Liu W, Feng F, Qu W.
      Recent studies suggested that Chotosan has ameliorative effects on vascular dementia through antioxidative pathways. Nevertheless, no systematic pharmacological research was conducted to evaluate the contribution of nuclear factor-E2-related factor 2 (Nrf2), a crucial regulator of antioxidative system, on Chotosan-induced neuroprotection invascular dementia. The present study aimed to investigate the neuroprotective effect of Chotosan on vascular dementia and reveal the possible molecular mechanism involving Nrf2. We found that Chotosan treatment could ameliorate memory impairment and reduce neuron cell loss induced by common carotid artery occlusion surgery. Furthermore, Chotosan could significantly reverse reactive oxygen species production, neuronal apoptosis and microglia over-activation in hippocampus. In addition, Chotosan enhanced Nrf2 expression and its nuclear translocation as well as its downstream antioxidant protein expression, NAD(P)H/quinone oxidoreductase 1 and heme oxygenase-1. These findings suggest that Chotosan exert neuroprotection in an animal model of vascular dementia via activating Nrf2-mediated antioxidant pathway. Chotosan may serve as a potential candidate and promising Nrf2 activator for treating vascular dementia.
    Keywords:  Chotosan; Cognitive deficits; Nrf2; Oxidative stress; Vascular dementia
    DOI:  https://doi.org/10.1016/j.jphs.2018.12.003
  34. Cell Stress Chaperones. 2019 Jan 17.
    Zhou J, Shi M, Li M, Cheng L, Yang J, Huang X.
      Sirtuin 3 (Sirt3)-modified mitochondrial fission participates in the progression of several types of cancers. However, its role in tongue cancer requires investigation. The aim of our study is to determine whether Sirt3 knockdown regulates the viability of tongue cancer cells via modulating mitochondrial fission. Two types of tongue cancer cells were used in the present study, and siRNA was transfected into the cells to suppress Sirt3 expression. Mitochondrial function and cell apoptosis were determined via immunofluorescence, Western blotting, ELISA, and qPCR assays. A pathway blocker was applied to verify the role of the JNK-Fis1 signaling pathway in regulation of mitochondrial fission. The present study showed that loss of Sirt3 promoted tongue cancer cell death in a manner dependent on mitochondrial apoptosis. Mitochondrial oxidative stress, energy metabolism disorder, mitochondrial cyt-c liberation, and mitochondrial apoptosis activation were observed after Sirt3 silencing. Furthermore, we demonstrated that Sirt3 knockdown activated mitochondrial stress via triggering Fis1-related mitochondrial fission and that inhibition of Fis1-related mitochondrial fission abrogated the pro-apoptotic effect of Sirt3 knockdown on tongue cancer cells. To this end, we found that Sirt3 modulated Fis1 expression via the c-Jun N-terminal kinases (JNK) signaling pathway and that blockade of the JNK pathway attenuated mitochondrial stress and repressed apoptosis in Sirt3 knockdown cells. Altogether, our results identified a tumor-suppressive role for Sirt3 deficiency in tongue cancer via activation of the JNK-Fis1 axis and subsequent initiation of fatal mitochondrial fission. Given these findings, strategies to repress Sirt3 activity and enhance the JNK-Fis1-mitochondrial fission cascade have clinical benefits for patients with tongue cancer.
    Keywords:  JNK-Fis1 signaling pathway; Mitochondrial dysfunction; Mitochondrial fission; Sirt3; Tongue cancer
    DOI:  https://doi.org/10.1007/s12192-019-00970-8
  35. PLoS One. 2019 ;14(1): e0210248
    Kim J, Pajarillo E, Rizor A, Son DS, Lee J, Aschner M, Lee E.
      Long-term exposure to elevated levels of manganese (Mn) causes manganism, a neurodegenerative disorder with Parkinson's disease (PD)-like symptoms. Increasing evidence suggests that leucine-rich repeat kinase 2 (LRRK2), which is highly expressed in microglia and macrophages, contributes to the inflammation and neurotoxicity seen in autosomal dominant and sporadic PD. As gene-environment interactions have emerged as important modulators of PD-associated toxicity, LRRK2 may also mediate Mn-induced inflammation and pathogenesis. In this study, we investigated the role of LRRK2 in Mn-induced toxicity using human microglial cells (HMC3), LRRK2-wild-type (WT) and LRRK2-knockout (KO) RAW264.7 macrophage cells. Results showed that Mn activated LRRK2 kinase by phosphorylation of its serine residue at the 1292 position (S1292) as a marker of its kinase activity in macrophage and microglia, while inhibition with GSK2578215A (GSK) and MLi-2 abolished Mn-induced LRRK2 activation. LRRK2 deletion and its pharmacological inhibition attenuated Mn-induced apoptosis in macrophages and microglia, along with concomitant decreases in the pro-apoptotic Bcl-2-associated X (Bax) protein. LRRK2 deletion also attenuated Mn-induced production of reactive oxygen species (ROS) and the pro-inflammatory cytokine TNF-α. Mn-induced phosphorylation of mitogen-activated protein kinase (MAPK) p38 and ERK signaling proteins was significantly attenuated in LRRK2 KO cells and GSK-treated cells. Moreover, inhibition of MAPK p38 and ERK as well as LRRK2 attenuated Mn-induced oxidative stress and cytotoxicity. These findings suggest that LRRK2 kinase activity plays a critical role in Mn-induced toxicity via downstream activation of MAPK signaling in macrophage and microglia. Collectively, these results suggest that LRRK2 could be a potential molecular target for developing therapeutics to treat Mn-related neurodegenerative disorders.
    DOI:  https://doi.org/10.1371/journal.pone.0210248
  36. Toxicol Mech Methods. 2019 Jan 14. 1-32
    Chen X, Peng X, Luo Y, You J, Yin D, Xu Q, He H, He M.
      Cardiotoxicity limits the clinical applications of doxorubicin (Dox), which mechanism might be excess generation of intracellular ROS. Quercetin (Que) is a flavonoid that possesses anti-oxidative activities, exerts myocardial protection. We hypothesized that the cardioprotection against Dox injury of Que involved 14-3-3γ, and mitochondria. To investigate the hypothesis, we treated primary cardiomyocytes with Dox and determined the effects of Que pretreatment with or without 14-3-3γ knockdown. We analyzed various cellular and molecular indexes. Our data showed that Que attenuated Dox-induced toxicity in cardiomyocytes by upregulating 14-3-3γ expression. Que pretreatment increased cell viability, SOD, catalase, and GPx activities, GSH levels, MMP and the GSH/GSSG ratio; decreased LDH and caspase-3 activities, MDA and ROS levels, mPTP opening and the percentage of apoptotic cells. However, Que's cardioprotection were attenuated by knocking down 14-3-3γ expression using pAD/14-3-3γ-shRNA. In conclusion, Que protects cardiomyocytes against Dox injury by suppressing oxidative stress, and improving mitochondrial function via 14-3-3γ.
    Keywords:  14-3-3γ; Quercetin; cardiotoxicity; doxorubicin; mitochondria; oxidative stress
    DOI:  https://doi.org/10.1080/15376516.2018.1564948
  37. Biochimie. 2019 Jan 09. pii: S0300-9084(19)30004-5. [Epub ahead of print]158 165-171
    Kim SH, Kil IS, Kwon OS, Kang BS, Lee DS, Lee HS, Lee JH, Park JW.
      The potent cytotoxicity of reactive oxygen species (ROS) can cause various diseases, however, it may also serve as a powerful chemotherapeutic strategy capable of killing cancer cells. Oxalomalate (OMA, α-hydroxy-β-oxalosuccinic acid), a tricarboxylic acid intermediate, is a well-known competitive inhibitor of two classes of NADP+-dependent isocitrate dehydrogenase (IDH) isoenzymes, which serve as the major antioxidants and redox regulators in the mitochondria and cytosol. In this study, we investigated the therapeutic effects of OMA in melanoma and elucidated the associated underlying mechanisms of action using in vitro and in vivo models. OMA targeting IDH enzymes suppressed melanoma growth through activation of apoptosis and inhibition of angiogenesis. Mechanistically, our findings showed that OMA activated p53-mediated apoptosis through ROS-dependent ATM-Chk2 signaling and reduced the expression of vascular endothelial growth factor through ROS-dependent E2F1-mediated hypoxia inducible factor-1α degradation. In particular, OMA-induced suppression of IDH activity resulted in induction of ROS stress response, ultimately leading to apoptotic cell death and antiangiogenic effects in melanoma cells. Thus, OMA might be a potential candidate drug for melanoma skin cancer therapy.
    Keywords:  Antiangiogenic effects; IDH; Oxalomalate; ROS
    DOI:  https://doi.org/10.1016/j.biochi.2019.01.004
  38. Cancer Manag Res. 2019 ;11 483-500
    Ray A, Jena S, Dash B, Sahoo A, Kar B, Patnaik J, Panda PC, Nayak S, Mahapatra N.
      Background: Hedychium coronarium Koen. (Zingiberaceae) is traditionally used as medicine in countries such as India, China, and Vietnam for treatment of various ailments including cancer. However, in spite of its implied significance in cancer treatment regimes, there are no reports so far involving the anticancerous attributes of H, coronarium ethanol extract (HCEE) on cancer cells and a more comprehensive study on its mechanism is still lacking.Materials and methods: The cytotoxicity of HCEE was evaluated by MTT and clonogenic survival assay. Annexin V/propidium iodide (PI), Hoechst 33342 staining, and TUNEL assay were performed to detect apoptosis. Cell cycle analysis was performed using PI staining. JC-1 and 2',7'-dichlorodihydrofluorescein diacetate assay were used to check the levels of MMP and ROS, respectively. Western blot analysis was carried out to measure the expression levels of proteins. Migration and invasion activity were assessed by wound healing and Transwell membrane assay, respectively.
    Results: Antiproliferative effect of HCEE was investigated in various cancerous and normal cell lines. Among these, HCEE significantly inhibited the survival of HeLa cells without affecting the viability of normal human umbilical vein endothelial cells. Annexin V/PI, Hoechst staining, and TUNEL assay showed HCEE induced apoptosis in HeLa cells in a dose-dependent manner. HCEE promoted cell cycle arrest at G1 phase in HeLa cells by upregulating the levels of p53 and p21 and downregulating the levels of cyclin D1, CDK-4, and CDK-6. Moreover, HCEE treatment upregulated the expression of Bax and downregulated the expression of Bcl-2. Additionally, HCEE activated the caspase cascade by increasing the activities of caspase-9, caspase-8, and caspase-3. The expression levels of Fas ligand and Fas were also upregulated. Further, HCEE inhibited the migratory potential of HeLa cells by downregulating MMP-2 and MMP-9 expression levels.
    Conclusion: Our results indicate H. coronarium exerts antiproliferative and apoptotic effects against HeLa cells, and therefore may be used for treatment against cervical cancer.
    Keywords:  HeLa cell line; Hedychium coronarium extract; apoptosis; cervical cancer; cytotoxicity
    DOI:  https://doi.org/10.2147/CMAR.S190004
  39. Cancer Chemother Pharmacol. 2019 Jan 16.
    Saad EA, Waly HM.
      PURPOSE: Searching for novel anticancer therapeutics which are effective and primarily less toxic is urgently needed. Drug encapsulation provides more protection of drug within the body with more stable drug circulation levels thus avoiding drug peak-related adverse effects. We aimed first to develop and characterize a nano-particulate drug delivery system using poly(lactic-co-glycolic acid) (PLGA) for the new compound N-butylpyridoquinoxaline 1,4-dioxide (NBPQD), and second to investigate its anticancer effect and the probable mechanism.METHODS: NBPQD-PLGA nano-particles were prepared and their shape, size, zeta potential, encapsulation efficiency (EE%), drug loading (DL%), drug release, anticancer activity against six human cancer cell lines, DNA binding ability, and flow cytometric analyses of apoptosis, cell cycle and caspase-3 activity were investigated.
    RESULTS AND CONCLUSIONS: NBPQD-PLGA nano-particles were spherical with diameter around 54 nm. Zeta potential, EE%, and DL% values were - 20.4 mV, 88% and 21.8%, respectively. Nano-particles exhibited higher marked anticancer activities (much lower IC50s) and changed the anticancer potency pattern towards all the studied cell lines compared to free NBPQD with superior potency against colorectal carcinoma (HCT-116, IC50 of 12.2 µg/mL). NBPQD-PLGA acts by induction of cancer cell apoptosis through oxidative stress, DNA damage, and activating a caspase-3 signaling pathway.
    Keywords:  Anticancer; Caspase-3; Cell cycle; Cytotoxicity; DNA damage; Quinoxaline
    DOI:  https://doi.org/10.1007/s00280-019-03770-0
  40. Exp Ther Med. 2019 Jan;17(1): 519-524
    Wang L, Gu W, Shi Y, Chen Y, Tan Y.
      The diaphragmatic fatigue that results from airflow obstruction is associated with the severe morbidity of patients with chronic obstructive pulmonary disease. Astragaloside IV (AS-IV) has antioxidant, anti-apoptotic and anti-inflammatory activities in various cell types. The present study aimed to evaluate the protective effects of AS-IV in diaphragmatic muscle cells. Diaphragmatic muscle cells extracted from neonatal rats were treated with a series of AS-IV concentrations (5, 10 or 20 mg/l) and the AKT inhibitor GSK690693 in the presence of interleukin-8 (IL-8). Cell proliferation and AKT phosphorylation were measured using Cell Counting Kit-8 and western blot assays, respectively. Cell apoptosis and reactive oxygen species (ROS) production were evaluated using flow cytometric analysis. Caspase activity and concentrations of proinflammatory factors (tumor necrosis factor-α, IL-6 and IL-8) were assessed using a caspase colorimetric assay and ELISA, respectively. IL-8 treatment resulted in decreased rates of cell proliferation and increased rates of AKT phosphorylation, cell apoptosis, caspase 3/9 activity, ROS production and proinflammatory factor production. AS-IV and GSK690693 treatment reversed the effects of IL-8. The effects of AS-IV were dose-dependent. The present results suggested that AS-IV is a candidate for the treatment of diaphragmatic fatigue due to its antioxidant, anti-apoptotic and anti-inflammatory activity.
    Keywords:  AKT; astragaloside IV; diaphragmatic fatigue; interleukin-8; oxidative stress
    DOI:  https://doi.org/10.3892/etm.2018.6940
  41. Exp Ther Med. 2019 Jan;17(1): 459-464
    Hao J, Zhao X, Yu W, Huang X, Huang Y.
      Renal fibrosis is a significant characteristic of chronic kidney diseases. Surfactant protein A (SP-A) is a recently identified fibrosis-associated factor in lung fibrosis; however, whether SP-A has the same role in renal fibrosis has remained elusive. The aim of the present study was to investigate the role of SP-A and its receptor calreticulin (CRT) in the pathogenesis of kidney fibrosis. The HK-2 human tubular epithelial cell line was cultured and treated with SP-A and SP-A + anti-CRT. The production of reactive oxygen species (ROS) at 30, 60 and 120 min was examined. Furthermore, cell apoptosis was assessed using an Annexin V assay and the expression of various proteins was measured using western blot analysis. In addition, the cell culture supernatants were collected and the expression of type I collagen was examined using ELISA. Compared with the control group, SP-A treatment significantly increased the ROS production, type I collagen secretion and cell apoptosis, which was partially inhibited by addition of anti-CRT. Furthermore, downregulation of matrix metalloproteinase (MMP)2 and -9 as well as upregulation of tissue inhibitor of metalloproteinase 1 indicated that SP-A treatment increased the degree of fibrosis in HK-2 cells, while addition of anti-CRT alleviated the fibrotic conditions. Finally, SP-A treatment significantly increased the expression of phosphorylated (p)-p38, p-p-65 and NADPH oxidase 2, which was partially inhibited by addition of anti-CRT. In conclusion, SP-A may participate in the pathogenesis of kidney fibrosis through binding to CRT and activate the mitogen-activated protein kinase/nuclear factor-κB-associated oxidative stress signaling pathway.
    Keywords:  MAPK; NF-κB; SP-A; calreticulin; kidney fibrosis; oxidative stress
    DOI:  https://doi.org/10.3892/etm.2018.6919
  42. Sci Rep. 2019 Jan 18. 9(1): 242
    Song HS, Song YH, Singh N, Kim H, Jeon H, Kim I, Kang SC, Chi KW.
      We report herein on the design, synthesis and biological activity of Ru-based self-assembled supramolecular bowls as a potent anticancer therapeutic in human hepatocellular cancer. The potent complex induces production of reactive oxygen species (ROS) by higher fatty acid β-oxidation and down-regulation of glucose transporter-mediated pyruvate dehydrogenase kinase 1 via reduced hypoxia-inducible factor 1α. Also, overexpressed acetyl-CoA activates the tricarboxylic acid cycle and the electron transport system and induces hypergeneration of ROS. Finally, ROS overexpressed through this pathway leads to apoptosis. Furthermore, we demonstrate that the naphthalene derived molecular bowl activates classical apoptosis via crosstalk between the extrinsic and intrinsic signal pathway. Our work into the mechanism of Ru-based self-assembled supramolecular bowls can provide valuable insight into the potential for use as a promising anticancer agent.
    DOI:  https://doi.org/10.1038/s41598-018-36755-9
  43. Photodiagnosis Photodyn Ther. 2019 Jan 12. pii: S1572-1000(18)30383-1. [Epub ahead of print]
    Li X, Bo X, Guo Y, Xiao Y, Xiao S.
      Neuroblastoma is the most common extracranial solid tumor of childhood. Advancements in treatments have improved survival rate of child suffering from this ailment. Novel therapeutic techniques may further reduce cancer related mortality. One of the several emerging therapeutic options is Photodynamic Therapy (PDT) that uses light activated photosensitizer (PS) inducing cell death by apoptosis and/or necrosis. Nanotechnology has contributed to improving photosensitizer for PDT, and increasing the efficiency of therapy using porphyrins and their derivatives. Efforts have been done to develop better mechanism to improve PS and consequently PDT effect. In this study, we investigated the efficacy of the PDT using porphyrins (TPOR) and TPOR/(CB[7])4 (TPOR: CB[7] = 1: 4). Here we report the PDT effect of TPOR and TPOR/(CB[7])4 in the treatment of the human neuroblastoma cell line (SH-SY5Y). The TPOR and TPOR/(CB[7])4 not show more significant dark-cytotoxicity and TPOR/(CB[7])4 had a more strong photodynamic effects than TPOR through generating reactive oxygen species (ROS) under irradiation with a 525 nm laser. The high photodynamic efficiency of TPOR/(CB[7])4 suggests that it has the potential to be a PDT agent.
    Keywords:  Cucurbit[7]uril; PDT; phototoxicity; porphyrins
    DOI:  https://doi.org/10.1016/j.pdpdt.2019.01.017
  44. J Exp Zool B Mol Dev Evol. 2019 Jan 17.
    Zhan C, Liu W, Hegazy AM, Zhang T, Kawan A, Zhang X.
      Obtaining oocytes from the adult female zebrafish (Danio rerio) ovary has enormous importance in the studies of developmental biology, toxicology, and genetics. It is vital to establish a simple and effective approach to ensure the quantity and quality of oocytes, which will enable the success of follow-up experimental investigation finally. Usually, oocytes are separated with mechanical or enzymatic methods, however, little studies have been done with concerns about the comparative effects. The present study separated zebrafish oocytes of Stage III with five frequently used methods, including stripping, pipetting, hyaluronidase (1.6 mg/ml), collagenase (0.4 mg/ml), and trypsin (0.1%). The cell viability, oxidative stress, mitogen-activated protein kinase (MAPK) protein phosphorylation, and apoptosis levels were selected as main biomarkers to evaluate the oocytes health status. The results showed that both trypsin and hyaluronidase isolation significantly upregulated germinal vesicle breakdown (GVBD) rates and downregulated p38 MAPK activity simultaneously. GVBD rates and survival rates were decreased notably in oocytes separated by the collagenase method. Above results indicate that zebrafish oocytes in vitro are sensitive to enzymatic treatments and the enzymatic isolation is not the suitable mean for collecting zebrafish oocytes although it is time-saving. The mechanical strategy of pipetting remarkably increased the reactive oxygen species and malondialdehyde level in isolated oocytes. Interestingly, oocytes separated with stripping show less physiological and biochemical damages. Therefore, stripping isolation is comparatively recommended as the optimum method for separating and collecting numerous intact and healthy zebrafish oocytes in vitro for the subsequent developmental research.
    Keywords:  GVBD; ROS; isolation; oocyte; stripping; viability
    DOI:  https://doi.org/10.1002/jez.b.22841
  45. Oncol Lett. 2019 Jan;17(1): 42-54
    Zhao H, Zhuang Y, Li R, Liu Y, Mei Z, He Z, Zhou F, Zhou Y.
      The present study examined the radiation biological response of cancer cells to different fractional irradiation doses and investigates the optimal fractional irradiation dose with improved biological effects. Radiobiological studies were performed at the molecular and cellular levels to provide insights into DNA damage and repair, and the apoptosis mechanism of cells that were exposed to different doses of X-ray irradiation (0, 2, 4, 6, 8, 10, 12.5, 15 and 20 Gy). Evidence of increased reactive oxygen species (ROS), DNA double strand breaks (DSB), cellular apoptosis, G2/M phase proportion and inhibition of cell proliferation were observed following irradiation. Differences in the ROS amount and apoptotic percentages of cells between the 2 and 4 Gy groups were insignificant. Compared with 0 Gy, the expression of the apoptosis suppression protein B-cell lymphoma-2 was decreased following at increased irradiation doses. However, apoptosis-associated protein Bcl-2-associated X (Bax), caspase-9 and BH3 interacting domain death agonist (Bid) were elevated following irradiation, compared with the control group (0 Gy). Furthermore, the expression levels of Bax in the 6, 8, 10 and 12.5 Gy groups were significantly increased, compared with the other groups. Caspase-9 expression with 2, 4, 6 and 8 Gy were increased compared with other groups, and the Bid levels with 6 and 8 Gy were also increased compared with other groups. G2/M phase arrest was associated with the increase of checkpoint kinase 1 and reduction of cyclin dependent kinase 1. DNA damage repair was associated with the protein Ku70 in the 2, 8, 10, 12.5, 15 and 20 Gy groups were less than other group. Compared with other group, Ku80 levels were reduced in the 6 and 8 Gy groups, and Rad51 levels were reduced in the 2, 8 and 10 Gy groups. The expression of hypoxia inducible factor-1α, c-Myc and glucose transporter 1 (GLUT1) demonstrated an increasing trend following irradiation in a dose-dependent manner, but the expression of pyruvate kinase M2, in the 2-10 Gy irradiation groups, and GLUT1, in the 12.5, 15 and 20 Gy irradiation groups, were reduced, compared with the other groups. Considering the DNA damage repair and apoptosis mechanisms at molecular and cellular levels, it was concluded that 2, 6, 8 and 10 Gy may be the optimal fractional dose that can promote cell apoptosis, and inhibit DNA damage repair and glycolysis.
    Keywords:  DNA damage repair; HeLa cells; apoptosis; fractional dose; glycolysis; irradiation
    DOI:  https://doi.org/10.3892/ol.2018.9566
  46. Vet Microbiol. 2019 Feb;pii: S0378-1135(18)30935-0. [Epub ahead of print]229 153-158
    Cardoso TC, Okamura LH, Baptistella JC, Borsanelli AC, Baptistiolli L, Ferreira HL, Gameiro R, Flores EF.
      Bovine alphaherpesvirus 1 (BHV1) and 5 (BHV5) are known to establish latent infections in sensory neurons of the trigeminal ganglion, yet leukocytes and tonsils have also been described as sites of latency in experimentally infected cattle. Little information is available on which immune cells are susceptible to BHV5 infection and how the infection may affect cell bioenergy. The aim of this study was to determine whether primary bovine monocyte-derived macrophages are susceptible to BHV5 infection and to evaluate parameters such as cell survival, virus replication and nitric oxide (NO) production. In addition, production of reactive oxygen species and mitochondrial damage were also analyzed. BHV5 infected cells underwent low rates of apoptosis but activated mitochondrial membrane depolarization and complex I. Additionally, production of high NO levels upon monocyte derived-macrophage infection did not interfere with the production of progeny virus. Overall, our findings revealed that primary cultures of bovine monocyte-derived macrophages support BHV5 replication in vitro and that mitochondrial dysfunction induced by infection apparently does not interfere with virus replication.
    Keywords:  BHV5; Cell bioenergy; Innate immune cells; Mitochondrion
    DOI:  https://doi.org/10.1016/j.vetmic.2019.01.004
  47. J Inorg Biochem. 2019 Jan 10. pii: S0162-0134(18)30653-6. [Epub ahead of print]193 1-8
    Yao Q, Qi J, Zheng Y, Qian K, Wei L, Maimaitiyiming M, Cheng Z, Wang Y.
      We synthesized five iron chelator derived from 2,6-diacetylpyridine bis(acylhydrazones) and proved their iron complexes structure by X-ray single crystal diffraction. These ligands have a significant anticancer proliferative activity and low cytotoxicity against normal cells. The Fe(III) complexes show reduced cytotoxic activity compared to the metal-free ligands. Anticancer mechanism studies indicate that ligands with a potential anticancer proliferation activity by inhibiting the activity of ribonucleotide reductase. Ligand rather than iron complexes regulate the expression of cell cycle associated proteins and inhibit cell cycle arrest in S phase. Apoptosis mechanism results showed that both ligand and iron complexes did not significantly promote apoptosis.
    Keywords:  Anticancer activity; Apoptosis; Cell cycle; Chelator; Reactive oxygen species
    DOI:  https://doi.org/10.1016/j.jinorgbio.2019.01.003
  48. ACS Nano. 2019 Jan 14.
    Li S, Liu R, Jiang X, Qiu Y, Song X, Huang G, Fu N, Lin L, Song J, Chen X, Yang H.
      The exploitation of gas therapy platforms holds great promise as a "green" approach for selective cancer therapy, however, it is often associated with some challenges, such as uncontrolled or insufficient gas generation and unclear therapeutic mechanisms. In this work, a gas therapy approach based on near-infrared (NIR) light triggered sulfur dioxide (SO2) generation was developed, and the therapeutic mechanism as well as in vivo anti-tumor therapeutic efficacy was demonstrated. A SO2 prodrug-loaded rattle-structured upconversion@silica nanoparticles (RUCSNs) was constructed to enable high loading capacity without obvious leakage, and to convert NIR light into ultraviolet (UV) light so as to activate the prodrug for SO2 generation. In addition, SO2 prodrug-loaded RUCSNs showed high cell uptake, good biocompatibility, intracellular tracking ability, and high NIR light triggered cytotoxicity. Furthermore, the cytotoxic SO2 was found to induce cell apoptosis accompanied with the increase of intracellular reactive oxygen species (ROS) levels and the damage of nuclear DNA. Moreover, efficient inhibition of tumor growth was achieved, associated with significantly prolonged survival of mice. Such NIR light-triggered SO2 therapy may provide an effective strategy to stimulate further development of synergistic cancer therapy platforms.
    DOI:  https://doi.org/10.1021/acsnano.8b08700
  49. Curr Pharm Des. 2019 Jan 18.
    Quintana J, Estévez F.
      Sesquiterpene lactones are naturally occurring compounds that have attracted considerable attention because to their vast array of biological activities. These plant-derived compounds contain the α-methylene-γ-butyrolactone functional group, which is the structural requirement for their pharmacological activities. Many of them exhibit cancer cell cytotoxicity and are promising anticancer agents through multiple mechanisms of action. Sesquiterpene lactones are alkylating agents that form covalent adducts in vivo and inhibit enzymes and key proteins. They are also potent apoptotic inducers in several cancer cells. This kind of cell death is recognized as a property that is useful for identifying anticancer drugs. This review focuses on the advances on cytotoxic sesquiterpene lactones and specifically the signal transduction pathways of cell death triggered in cancer cells.
    Keywords:  Apoptosis; Caspases; Cell cycle arrest; Cell death; Cytotoxicity; Mitogen-activated protein kinases; Reactive oxygen species.; Sesquiterpene lactones
    DOI:  https://doi.org/10.2174/1381612825666190119114323
  50. Mol Neurobiol. 2019 Jan 16.
    Zhang J, Feng J, Ma D, Wang F, Wang Y, Li C, Wang X, Yin X, Zhang M, Dagda RK, Zhang Y.
      Protein kinase A (PKA) is a ser/thr kinase that is critical for maintaining essential neuronal functions including mitochondrial homeostasis, bioenergetics, neuronal development, and neurotransmission. The endogenous pool of PKA is targeted to the mitochondrion by forming a complex with the mitochondrial scaffold A-kinase anchoring protein 121 (AKAP121). Enhanced PKA signaling via AKAP121 leads to PKA-mediated phosphorylation of the fission modulator Drp1, leading to enhanced mitochondrial networks and thereby blocking apoptosis against different toxic insults. In this study, we show for the first time that AKAP121/PKA confers neuroprotection in an in vitro model of oxidative stress induced by exposure to excess glutamate. Unexpectedly, treating mouse hippocampal progenitor neuronal HT22 cells with an acute dose or chronic exposure of glutamate robustly elevates PKA signaling, a beneficial compensatory response that is phenocopied in HT22 cells conditioned to thrive in the presence of excess glutamate but not in parental HT22 cells. Secondly, redirecting the endogenous pool of PKA by transiently transfecting AKAP121 or transfecting a constitutively active mutant of PKA targeted to the mitochondrion (OMM-PKA) or of an isoform of AKAP121 that lacks the KH and Tudor domains (S-AKAP84) are sufficient to significantly block cell death induced by glutamate toxicity but not in an oxygen deprivation/reperfusion model. Conversely, transient transfection of HT22 neuronal cells with a PKA-binding-deficient mutant of AKAP121 is unable to protect against oxidative stress induced by glutamate toxicity suggesting that the catalytic activity of PKA is required for AKAP121's protective effects. Mechanistically, AKAP121 promotes neuroprotection by enhancing PKA-mediated phosphorylation of Drp1 to increase mitochondrial fusion, elevates ATP levels, and elicits an increase in the levels of antioxidants GSH and superoxide dismutase 2 leading to a reduction in the level of mitochondrial superoxide. Overall, our data supports AKAP121/PKA as a new molecular target that confers neuroprotection against glutamate toxicity by phosphorylating Drp1, to stabilize mitochondrial networks and mitochondrial function and to elicit antioxidant responses.
    Keywords:  A-kinase anchoring protein 121; Dynamin-related protein 1 (Drp1); HT22; Mitochondrion; Oxidative stress; Protein kinase A
    DOI:  https://doi.org/10.1007/s12035-018-1464-3
  51. Evid Based Complement Alternat Med. 2018 ;2018 2545024
    Cheng Y, Song J, Hu P, Zhu Y.
      Tetrahydroxystilbene glucoside (TSG) is extracted from a famous Chinese herbal medicine which is widely used as an antiaging agent in history. Lots of studies gave evidence that TSG exhibited benefits to brain, like improvement of learning and memory and synaptic plasticity. Moreover, the polyphenolic structure of TSG enables its capability to prevent cerebral ischemia/reperfusion injury (IRI) by reducing apoptosis and ROS/RNS generation. Due to its antioxidant profile, TSG had been demonstrated to alleviate cardiac toxicity by regulating biochemical indexes and ROS. However, whether TSG exhibited cardioprotective effects via mitochondrial energy metabolic functions, which played crucial role in IRI, remained unclear. Here, we used an in vitro aging model of cardiomyocytes to evaluate the effects of TSG on transient hypoxia-pretreated hypoxia/reoxygenation (H/R) injury and mitochondrial energy metaolism. Our results showed that TSG enhanced cardioprotective effect of transient hypoxia on H/R by reducing excessive ROS production and calcium overloading. Significant improvements of mitochondrial respiratory functions and ketone body metabolism elucidated that TSG restored the effect of transient hypoxia on H/R injury in aging cardiomyocytes via upregulating mitochondrial energy metabolism.
    DOI:  https://doi.org/10.1155/2018/2545024
  52. PLoS One. 2019 ;14(1): e0210513
    Wei PL, Huang CY, Chang YJ.
      The poor prognosis of hepatocellular carcinoma (HCC) has been attributed to a high frequency of tumor metastasis and recurrence even after successful surgical resection. With less than 30% of patients benefiting from curative treatment, alternative treatment regimens for patients with advanced HCC are needed. Propyl gallate (PG), a synthetic antioxidant used in preserving food and medicinal preparations, has been shown to induce cancer cell death, but the anticancer effects of PG in HCC are unclear. In the present study, we demonstrated that PG inhibited HCC cell proliferation in vitro and in zebrafish models in vivo in a dose- and time-dependent manner. PG also induced cell apoptosis and increased the number of necrotic cells in a time- and dose-dependent manner as determined using a high-content analysis system. We found that PG also increased the intracellular levels of superoxide and reactive oxidative stress as well as the formation of autophagosomes and lysosomes. Regarding the molecular mechanism, PG did not alter the levels of autophagy-related 5 (ATG5), ATG5/12 or Beclin-1 but increased the rate of the LC3-I to LC3-II conversion, suggesting autophagy induction. PG exposure increased the levels of the pro-apoptotic proteins cleaved caspase-3, cleaved PARP, Bax, and Bad and a decreased level of the anti-apoptotic protein Bcl-2. In conclusion, we demonstrate that PG inhibits HCC cell proliferation through enhanced ROS production and autophagy activation. Finally, PG-treated cells induced cell apoptosis and may be a new candidate for HCC therapy.
    DOI:  https://doi.org/10.1371/journal.pone.0210513
  53. Cancer Sci. 2019 Jan 18.
    Gu C, Luo J, Lu X, Tang Y, Ma Y, Yun Y, Cao J, Cao J, Huang Z, Zhou X, Zhang S.
      Radiotherapy has been widely used for the clinical management of esophageal squamous cell carcinoma. However, radioresistance remains a serious concern that prevents the efficacy of ESCC radiotherapy. REV7, the structural subunit of eukaryotic DNA polymerase ζ, has multiple functions in bypassing DNA damage and modulating mitotic arrest in human cell lines. However, the expression and molecular function of REV7 in ESCC progression remains unclear. In this study, we first examined the expression of REV7 in clinical ESCC samples, and we found higher expression of REV7 in ESCC tissues compared to matched adjacent or normal tissues. Knockdown of REV7 resulted in decreased colony formation and increased apoptosis in irradiated Eca-109 and TE-1 cells coupled with decreased tumor weight in a xenograft nude mouse model post-irradiation. Conversely, overexpression of REV7 resulted in the radioresistance in vitro and in vivo. Moreover, silencing of REV7 induced increased reactive oxygen species (ROS) levels post irradiation. To elucidate the underlying mechanism, proteomic analysis of REV7-interacting proteins revealed that REV7 interacted with peroxiredoxin 2 (PRDX2), a well-known antioxidant protein. Existence of REV7-PRDX2 complex and its augmentation post irradiation were further validated by immunoprecipitation and immunofluorescence assays. REV7 knockdown significantly disrupted the presence of nuclear PRDX2 post irradiation, which resulted in oxidative stress. REV7-PRDX2 complex also assembled onto DNA double strand breaks (DSBs), whereas REV7 knockdown evidently increased DSBs that were unmerged by PRDX2. Taken together, this study sheds light on REV7-modulated radiosensitivity through interacting with PRDX2, which provides a novel target for ESCC radiotherapy. This article is protected by copyright. All rights reserved.
    Keywords:  DNA double strand breaks; PRDX2; REV7; radioresistance; reactive oxygen species
    DOI:  https://doi.org/10.1111/cas.13946
  54. Am J Physiol Renal Physiol. 2019 Jan 16.
    Tao M, Shi Y, Tang L, Wang Y, Fang L, Jiang W, Lin T, Qiu A, Zhuang S, Liu N.
      Extracellular signal-regulated kinases 1 and 2 (ERK1/2) are serine/threonine kinases and function as regulators of cellular proliferation and differentiation. Recently, we demonstrated that inhibition of ERK1/2 alleviates the development and progression of hyperuricemia nephropathy (HN). However, its potential roles in uric acid-induced tubular epithelial-mesenchymal transition (EMT) and tubular epithelial cell injury are unknown. In this study, we showed that hyperuricemic injury induced EMT as characterized by down-regulation of E-cadherin and up-regulation of Vimentin and Snail1 in a rat model of HN. This was coincident with epithelial cells arrested at the G2/M phase of cell cycle, activation of Notch1/Jagged-1 and Wnt/β-catenin signaling pathways, and upregulation of matrix metalloproteinases 2 (MMP2) and MMP9. Administration of U0126, a selective inhibitor of ERK1/2, blocked all these responses. U0126 was also effective in inhibiting renal tubular cell injury as shown by decreased expression of lipocalin-2 (Lcn2) and kidney injury molecule-1 (Kim-1) and active forms of caspase-3. U0126 or ERK1/2 siRNA can inhibit tubular cell EMT and cell apoptosis as characterized with decreased expression of cleaved caspase-3. Moreover, ERK1/2 inhibition suppressed hyperuricemic injury-induced oxidative stress as indicated by decreased malondialdehyde (MDA) and increased superoxide dismutase (SOD). Collectively, ERK1/2 inhibition-elicited renal protection is associated with inhibition of EMT through inactivation of multiple signaling pathways and matrix metalloproteinases, as well as attenuation of renal tubule injury by enhancing cellular resistance to oxidative stress.
    Keywords:  ERK1/2; cell apoptosis; epithelial-mesenchymal transition; hyperuricemic nephropathy; oxidative stress
    DOI:  https://doi.org/10.1152/ajprenal.00480.2018
  55. J Biomed Mater Res A. 2019 Jan 16.
    Liu CG, Kankala RK, Liao HY, Chen AZ, Wang SB.
      Spurred by the current advancements in engineering various intelligent nanoparticle-based drug delivery systems, conjugation of drugs with the stimuli-responsive molecular switches has become one of the most efficient approaches to deliver a drug cargo in spatiotemporal controlled fashions. In this study, we fabricated an innovative pH-triggered Hydrazone-carboxylate complex of doxorubicin (Dox), which was subsequently encapsulated in the layered double hydroxide (LDH) nanoparticles for effective cancer therapeutics. These two-dimensional (2D) biodegradable matrices efficiently delivered Dox by pH-triggered release in the acidic lysosomal environment and their subsequent escape to cytosol. Moreover, the delivered Dox molecules and high positively-charged surfaces of LDHs facilitated the cancer cell ablation via enhancing the cathepsins-mediated cell apoptosis assisted by free radical species generation. The critical advancements in the nanoparticle-based designs and substantial ablation of tumor cells through a free radical attack indicate that the designed pH-triggered drug composites can be used for efficient cancer therapeutics. This article is protected by copyright. All rights reserved.
    Keywords:  Cathepsin-mediated effect; Doxorubicin; Ion-exchange; Layered double hydroxides; pH-responsive release
    DOI:  https://doi.org/10.1002/jbm.a.36610
  56. Amino Acids. 2019 Jan 17.
    Jafri AJA, Agarwal R, Iezhitsa I, Agarwal P, Ismail NM.
      This study aimed to evaluate effect of TAU on NMDA-induced changes in retinal redox status, retinal cell apoptosis and retinal morphology in Sprague-Dawley rats. Taurine was injected intravitreally as pre-, co- or post-treatment with NMDA and 7 days post-treatment retinae were processed for estimation of oxidative stress, retinal morphology using H&E staining and retinal cell apoptosis using TUNEL staining. Treatment with TAU, particularly pre-treatment, significantly increased retinal glutathione, superoxide dismutase and catalase levels compared to NMDA-treated rats; whereas, the levels of malondialdehyde reduced significantly. Reduction in retinal oxidative stress in TAU pre-treated group was associated with significantly greater fractional thickness of ganglion cell layer within inner retina and retinal cell density in inner retina. TUNEL staining showed significantly reduced apoptotic cell count in TAU pre-treated group compared to NMDA group. It could be concluded that TAU protects against NMDA-induced retinal injury in rats by reducing retinal oxidative stress.
    Keywords:  NMDA; Oxidative stress; Retina; Taurine
    DOI:  https://doi.org/10.1007/s00726-019-02696-4
  57. Sci Rep. 2019 Jan 17. 9(1): 195
    Riahi-Chebbi I, Souid S, Othman H, Haoues M, Karoui H, Morel A, Srairi-Abid N, Essafi M, Essafi-Benkhadir K.
      Resistance to 5-Fluorouracil chemotherapy is a major cause of therapeutic failure in colon cancer cure. Development of combined therapies constitutes an effective strategy to inhibit cancer cells and prevent the emergence of drug resistance. For this purpose, we investigated the anti-tumoral effect of thirteen phenolic compounds, from the Tunisian quince Cydonia oblonga Miller, alone or combined to 5-FU, on the human 5-FU-resistant LS174-R colon cancer cells in comparison to parental cells. Our results showed that only Kaempferol was able to chemo-sensitize 5-FU-resistant LS174-R cells. This phenolic compound combined with 5-FU exerted synergistic inhibitory effect on cell viability. This combination enhanced the apoptosis and induced cell cycle arrest of both chemo-resistant and sensitive cells through impacting the expression levels of different cellular effectors. Kaempferol also blocked the production of reactive oxygen species (ROS) and modulated the expression of JAK/STAT3, MAPK, PI3K/AKT and NF-κB. In silico docking analysis suggested that the potent anti-tumoral effect of Kaempferol, compared to its two analogs (Kaempferol 3-O-glucoside and Kampferol 3-O-rutinoside), can be explained by the absence of glucosyl groups. Overall, our data propose Kaempferol as a potential chemotherapeutic agent to be used alone or in combination with 5-FU to overcome colon cancer drug resistance.
    DOI:  https://doi.org/10.1038/s41598-018-36808-z