bims-aporos Biomed news
on Apoptosis and reactive oxygen species
Issue of 2018‒12‒23
forty papers selected by
Gavin McStay
Staffordshire University


  1. Int J Mol Med. 2018 Dec 12.
    Zhao H, Wang R, Ye M, Zhang L.
      Oxidative stress serves a vital function in the pathogenesis of age‑related macular degeneration (AMD); genipin (GP) possesses antioxidative properties. The present study aimed to investigate the effects of GP on retinal pigment epithelial (RPE) cells induced by H2O2 and the underlying mechanism. ARPE‑19 cells were subjected to H2O2 treatment to induce oxidative damage. Cell viability was determined via an MTT assay. Reactive oxygen species (ROS) levels and cell apoptosis were detected by flow cytometry. Nuclear factor‑erythroid 2‑related factor‑2 (Nrf2) signaling‑associated and the expression of apoptosis‑associated factors were measured using reverse transcription‑quantitative polymerase chain reaction assay and western blotting. The results revealed that 200 µM H2O2 and 30 µM GP were determined to be the optimal concentrations for subsequent experimentation. GP reversed the inhibitory effects of H2O2 by promoting cell viability, attenuating ROS accumulation and cell apoptosis, and increased the expression of Nrf2, heme oxygenase‑1 (HO‑1) and NAD(P)H: Quinine oxidoreductase 1 (NQO1); Nrf2 silencing inhibited HO‑1 and NQO1 expression. In addition, Nrf2 silencing enhanced the effects of H2O2 by promoting ROS production and cell apoptosis. Compared with H2O2, Nrf2 silencing further decreased the expression levels of B‑cell lymphoma‑2 (Bcl‑2), but increased that of Bcl‑2‑associated X protein and cleaved‑caspase‑3. The results of the present study revealed that Nrf2 silencing attenuated the protective effects of GP on H2O2‑induced injury in ARPE‑19 cells by promoting apoptosis and oxidation. Collectively, GP attenuated oxidative damage induced by H2O2 in ARPE‑19 cells. Furthermore, the molecular mechanism may be associated with the Nrf2 signaling pathway. The findings of the present study nay provide insight into a potential therapeutic agent for the treatment of AMD.
    DOI:  https://doi.org/10.3892/ijmm.2018.4027
  2. Cell Physiol Biochem. 2018 Dec 14. 51(6): 2938-2954
    Shen J, Cao S, Sun X, Pan B, Cao J, Che D, Jin S, Cao Y, Tian Y, Yu Y.
      BACKGROUND/AIMS: Sonodynamic therapy (SDT) is expected to be a new method to solve the clinical problems caused by advanced metastasis in patients with lung cancer. The use of ultrasound has the advantage of being noninvasive, with deep-penetration properties. This study explored the anti-tumor effect of SDT with a new sonosensitizer, sinoporphyrin sodium (DVDMS), on the human small cell lung cancer H446 cell line in vitro and in vivo.METHODS: Absorption of DVDMS was detected by a fluorescence spectrophotometer, and DVDMS toxicity was determined using a Cell Counting Kit-8. Mitochondrial membrane potential (MMP) was assessed using the JC-1 fluorescent probe. Cell apoptosis was measured by flow cytometry, and apoptosis-related proteins were detected by western blotting. The expression of cytokines was measured using an enzyme-linked immunosorbent assay and quantitative real-time PCR. To verify the in vitro results, we detected tumor volumes and weight changes in a xenograft nude mouse model after DVDMS-SDT. Hematoxylin and eosin staining was used to observe changes to the tumor, heart, liver, spleen, lung, and kidney of the mice, and immunohistochemistry was used to examine changes in the expression of tumor CD34 and receptor-interacting protein kinase-3 (RIP3), while terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling was used to observe apoptosis in tumor tissues.
    RESULTS: DVDMS-SDT-treated H446 cells increased the rate of cellular apoptosis and the levels of reactive oxygen species (ROS), cleaved caspase-3, cleaved caspase-8, cleaved caspase-9, and caspase-10, and decreased the levels of MMP, RIP3, B-cell lymphoma 2, vascular endothelial growth factor, and tumor necrosis factor-α. The sonotoxic effect was mediated by ROS and was reduced by a ROS scavenger (N-acetyl-L-cysteine). In the in vivo mouse xenograft model, DVDMS-SDT showed efficient anti-cancer effects with no visible side effects.
    CONCLUSION: DVDMS-SDT induced apoptosis in H446 cells, in part by targeting mitochondria through the mitochondria-mediated apoptosis signaling pathway, and the extrinsic apoptosis pathway was also shown to be involved. Both apoptosis and changes in RIP3 expression were closely related to the generation of ROS. DVDMS-SDT will be advantageous for the management of small cell lung cancer due to its noninvasive characteristics.
    Keywords:  Apoptosis; H446 cells; Reactive oxygen species; Sinoporphyrin sodium; Sonodynamic therapy
    DOI:  https://doi.org/10.1159/000496045
  3. Iran J Pharm Res. 2018 ;17(4): 1476-1487
    Aghvami M, Eshghi P, Zarei MH, Arefi H, Sattari F, Zarghi A, Pourahmad J.
      B-acute lymphoblastic leukemia (B-ALL) is the frequent pediatric malignity. Chemotherapy is the most practical approaches to deal with such malignancies. Microtubule-targeted agents are one of the most strategic drugs which formerly used in chemotherapy. Although colchicine-binding anti-tubulin agents exhibited promising effects in clinical trials, their exact mechanism of action is not fully understood. In this study, the effects of two newly synthesized of colchicine derivatives were investigated on cell viability of cancerous and normal lymphocytes. The viability test was carried out by MTT assay. Apoptosis vs. necrosis was measured by double staining with annexin V/PI, and caspase-3 as the ultimate mediator of apoptotic measured through the colorimetric assay. Parameters of mitochondrial damage (ROS formation, MMP (Mitochondrial Membrane Potential) decline, mitochondrial swelling, and cytochrome c release following treatment by colchicine derivatives. By focusing on mitochondrial parameters, we showed that following treatment by two newly synthesized colchicine derivatives, apoptosis is triggered in cancerous B-lymphocytes. We demonstrated these compounds could activate apoptosis in cancerous lymphocytes by augmentation of reactive oxygen species (ROS), a decline in mitochondrial membrane potential (MMP), mitochondrial swelling, release of cytochrome c, and also caspase-3 activation. Considering the obtained evidence, these inhibitors could be the new therapeutic strategies in ALL treatment.
    Keywords:  Apoptosis; B-acute lymphoblastic leukemia; Caspase cascade; Microtubules inhibitors; Mitochondria; VDAC
  4. Biochimie. 2018 Dec 18. pii: S0300-9084(18)30354-7. [Epub ahead of print]
    Mai HN, Sharma G, Sharma N, Shin EJ, Kim DJ, Pham DT, Trinh QD, Jang CG, Nah SY, Jeong JH, Kim HC.
      Cocaine, an addictive drug, is known to induce hepatotoxicity via oxidative damage and proapoptosis. Since p53, a tumor suppressor gene, plays a major role in inducing oxidative stress and apoptosis, we examined the role of p53 inhibition against cocaine-induced hepatotoxicity. Cocaine treatment significantly increased oxidative parameters (i.e., reactive oxygen species, 4-hydroxylnonenal, and protein carbonyl) in the liver of wild type (WT) mice. We found that the pharmacological (i.e. pifithrin-α) and genetic (i.e. p53 knockout) inhibition of p53 significantly attenuates cocaine-induced hepatotoxicity. Cocaine treatment increased alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels in the serum of mice, signifying hepatic damage. Consistently, these increases were attenuated by inhibition of p53, implying protection against cocaine-induced hepatic damage. In addition, cocaine treatment significantly increased PKCδ, cleaved PKCδ and p53 levels in the liver of WT mice. These increases were followed by the interaction between p53 and PKCδ, and pro-apoptotic consequences (i.e., cytosolic release of cytochrome c, activation of caspase-3, increase in Bax level and decreases in Bcl-2 and Bcl-xL levels). These changes were attenuated by p53 depletion, reflecting that the critical role of PKCδ in p53-mediated apoptotic potentials. Combined, our results suggest that the inhibition of p53 is important for protection against oxidative burdens, pro-apoptotic events, and hepatic degeneration induced by cocaine.
    Keywords:  Cocaine; Liver; Oxidative stress; Pro-apoptosis; Protein kinase Cδ; p53 knockout mice
    DOI:  https://doi.org/10.1016/j.biochi.2018.12.009
  5. Toxicol In Vitro. 2018 Dec 18. pii: S0887-2333(18)30317-5. [Epub ahead of print]
    Shabalala SC, Dludla PV, Muller CJF, Nxele X, Kappo AP, Louw J, Johnson R.
      Aspalathin (ASP) is a C-dihydrochalcone abundantly found in Aspalathus linearis. While we have provide evidence that ASP can protect H9c2 cardiomyoblasts against doxorubicin (Dox)-induced apoptosis through regulation of autophagy, the complete mechanism involved in the cardioprotective effect of this dihydrochalcone remains to be explored. Here we provide evidence that ASP reverses Dox-induced apoptosis through the amelioration of oxidative stress in H9c2 cardiomyoblasts. Cultured cells were treated with 0.2 μM Dox or co-treated with either 20 μM dexrazoxane (Dexra) or 0.2 μM ASP daily for five days, to a final dose of 1 μM Dox, 100 μM Dexra and 1 μM ASP, respectively. Superoxide dismutase, catalase, glutathione, malondialdehyde and dichloro-dihydro-fluorescein diacetate fluorescence were used as end-point measurements for oxidative stress, while JC-1 and TUNEL labeling were performed to assess mitochondria depolarization and apoptosis. Co-treatment with ASP attenuated Dox-induced cardiotoxicity by improving endogenous antioxidant levels and mitochondrial membrane potential, while inhibiting reactive oxygen species production and cellular apoptosis. These findings suggested that ASP can prevent Dox-induced oxidative stress and apoptosis and needs further assessment to confirm its therapeutic potential to prevent Dox-induced cardiotoxicity.
    Keywords:  Apoptosis; Aspalathin; Cardiotoxicity; Doxorubicin; Oxidative stress
    DOI:  https://doi.org/10.1016/j.tiv.2018.12.012
  6. Molecules. 2018 Dec 19. pii: E3372. [Epub ahead of print]23(12):
    Shen YH, Wang LY, Zhang BB, Hu QM, Wang P, He BQ, Bao GH, Liang JY, Wu FH.
      Ethyl rosmarinate (RAE) is one of the active constituents from Clinopodium chinense (Benth.) O. Kuntze, which is used for diabetic treatment in Chinese folk medicine. In this study, we investigated the protective effect of RAE on high glucose-induced injury in endothelial cells and explored its underlying mechanisms. Our results showed that both RAE and rosmarinic acid (RA) increased cell viability, decreased the production of reactive oxygen species (ROS), and attenuated high glucose-induced endothelial cells apoptosis in a dose-dependent manner, as evidenced by Hochest staining, Annexin V⁻FITC/PI double staining, and caspase-3 activity. RAE and RA both elevated Bcl-2 expression and reduced Bax expression, according to Western blot. We also found that LY294002 (phosphatidylinositol 3-kinase, or PI3K inhibitor) weakened the protective effect of RAE. In addition, PDTC (nuclear factor-κB, or NF-κB inhibitor) and SP600125 (c-Jun N-terminal kinase, or JNK inhibitor) could inhibit the apoptosis in endothelial cells caused by high glucose. Further, we demonstrated that RAE activated Akt, and the molecular docking analysis predicted that RAE showed more affinity with Akt than RA. Moreover, we found that RAE inhibited the activation of NF-κB and JNK. These results suggested that RAE protected endothelial cells from high glucose-induced apoptosis by alleviating reactive oxygen species (ROS) generation, and regulating the PI3K/Akt/Bcl-2 pathway, the NF-κB pathway, and the JNK pathway. In general, RAE showed greater potency than RA equivalent.
    Keywords:  apoptosis; ethyl rosmarinate; high glucose; rosmarinic acid; vascular endothelial cell
    DOI:  https://doi.org/10.3390/molecules23123372
  7. Chemosphere. 2018 Dec 08. pii: S0045-6535(18)32383-X. [Epub ahead of print]219 517-525
    Jiang J, Lv L, Wu S, An X, Wang F, Liu X, Zhao X.
      Kresoxim-methyl (KM) is a broad spectrum strobilurin fungicide that has been used widely on crops around the world. In the present study, we aimed to investigate the toxic effects of KM using various sublethal endpoints during zebrafish (Danio rerio) larval development. Results showed that the LC50 values of KM to zebrafish at multiple life stages (embryo, larvae, juvenile and adult) were 0.340, 0.224, 0.328 and 0.436 mg/L, respectively. The transcription patterns of 45 genes involved in hypothalamic-pituitary-thyroid/gonadal (HPT/HPG) axis, oxidative stress and apoptosis revealed KM could affect zebrafish larval development at multiple pathways. The activities of aromatase, superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPX), caspase 3 (Cas3) and caspase 9 (Cas9), and the levels of estradiol (E2), vitellogenin (VTG), thyroid hormones (T3 and T4), reactive oxygen species (ROS) and ATP after embryos exposed to KM for 3 d, 6 d and 10 d were correlated well with the transcription of the corresponding molecules involved in these pathways. In addition to providing the first description of the toxic effects induced by KM during larval development, the results of present study also provided the potential mechanisms of KM on multi-level biomarker responses in larval zebrafish.
    Keywords:  Apoptosis; Endocrine disruption; Kresoxim-methyl; Oxidative stress; Zebrafish
    DOI:  https://doi.org/10.1016/j.chemosphere.2018.12.061
  8. Chem Biol Interact. 2018 Dec 18. pii: S0009-2797(18)30977-3. [Epub ahead of print]
    Saavedra E, Del Rosario H, Brouard I, Quintana J, Estévez F.
      In this study, we investigated the effects of synthetic 6'-benzyloxy-4-bromo-2'-hydroxychalcone on viabilities of seven human leukaemia cells. It was cytotoxic against U-937, HL-60, K-562, NALM-6, MOLT-3 cells, and also against Bcl-2-overexpressing U-937/Bcl-2 cells and P-glycoprotein-overexpressing K-562/ADR, but had no significant cytotoxic effects against quiescent or proliferating human peripheral blood mononuclear cells. This chalcone is a potent apoptotic inducer in human leukaemia U-937 cells. Cell death was (i) mediated by the activation and the cleavage of initiator and executioner caspases and poly(ADP-ribose) polymerase; (ii) prevented by the pan-caspase inhibitor z-VAD-fmk, and by the selective caspase-3/7, -6 and -8 inhibitors, and by a cathepsins B/L inhibitor; (iii) associated with the release of mitochondrial proteins, including cytochrome c and Smac/DIABLO; (iv) accompanied by dissipation of the mitochondrial membrane potential, (v) partially blocked by the inhibition of p38MAPK and (vi) mostly abrogated by catalase. In conclusion, the synthetic chalcone is cytotoxic against several types of human leukaemia cell with apoptosis being induced by activation of the extrinsic pathway and the generation of reactive oxygen species.
    Keywords:  Apoptosis; Caspases; Chalcone; Cytotoxicity
    DOI:  https://doi.org/10.1016/j.cbi.2018.12.010
  9. Mol Med Rep. 2018 Dec 12.
    Li J, Zheng J.
      Theaflavins (TFs) are the main bioactive polyphenols in tea and contribute to protection against oxidative stress. Excessive reactive oxygen species (ROS) accumulation can lead to the disruption of cartilage homeostasis. The present study examined the potential effects of TFs on H2O2‑induced cartilage degeneration in vitro. Cell Counting kit (CCK‑8) was used to determine cell viability, and flow cytometric analysis was used to detect ROS, apoptosis and DNA damage. Reverse transcription‑quantitative polymerase chain reaction (RT‑qPCR) and western blotting were used to detect the expression levels of target factors. The present study revealed that TFs effectively reduced the expression of catabolic factors, including matrix metalloproteinase‑13, interleukin‑1 and cartilage glycoprotein 39. TFs inhibited ROS generation in cartilage degeneration, and suppressed apoptosis and DNA damage caused by oxidative stress. TFs also downregulated the expression levels of cleaved caspase‑3 and B‑cell lymphoma 2‑associated X protein, and the DNA damage‑related genes, ATR serine/threonine kinase and ATM serine/threonine kinase. Furthermore, TFs enhanced the activity of glutathione peroxidase 1 and catalase, but reduced the expression levels of phosphorylated (p)‑AKT serine/threonine kinase (AKT) and p‑Forkhead box O3 (FOXO3)a. Conversely, the effects of TFs on apoptosis and DNA damage were reversed by persistent activation of AKT. In conclusion, TFs prevented cartilage degeneration via AKT/FOXO3 signaling in vitro. The present study suggested that TFs may be a potential candidate drug for the prevention of cartilage degeneration.
    DOI:  https://doi.org/10.3892/mmr.2018.9745
  10. Biol Trace Elem Res. 2018 Dec 18.
    Peng W, Xu S, Zhang J, Zhang Y.
      Increasing evidence has suggested an important role played by reactive oxygen species (ROS) in the pathogenesis of fluorosis. Accumulating evidence demonstrates that vitamin C administration ameliorate sodium fluoride (NaF)-induced oxidative stress. However, the potentially beneficial effects of vitamin C against NaF-induced cytotoxicity and the underlying molecular mechanisms of this protection are not fully understood. Here, we found that NaF stimulated cytotoxicity, increased mitochondrial reactive oxygen species (mROS) production, and induced apoptosis in F9 embryonic carcinoma cells. Consistent with this finding, NaF exposure was associated with decreased Sirtuin 1 (Sirt1) protein expression, thus promoted the acetylation of manganese superoxide dismutase (SOD2), a key enzyme involved in regulating mROS production. However, all NaF-induced mitochondrial oxidative injuries were efficiently ameliorated by overexpression of Sirt1 or incubation with Mito-TEMPO (a SOD2 mimetic). Moreover, pretreatment with vitamin C enhanced the expression of Sirt1 and decreased NaF-induced mitochondrial oxidative stress and apoptosis. Knockdown of Sirt1 blocked the vitamin C-mediated reduction in mROS and apoptosis via inhibiting Sirt1-SOD2 signaling. Importantly, sodium-dependent vitamin C transporter 2 (SVCT-2) siRNA was found to partially block the ability of vitamin C to promote Sirt1/SOD2 signaling. In summary, our data indicate that Sirt1 plays a pivotal role in the ability of vitamin C to stimulate SOD2 activity and attenuate mitochondrial oxidative stress, which partially through vitamin C receptor in NaF-induced F9 cells injury.
    Keywords:  F9 cells; NaF; SOD2; SVCT-2; Sirt1; Vitamin C
    DOI:  https://doi.org/10.1007/s12011-018-1599-0
  11. J Gastroenterol Hepatol. 2018 Dec 21.
    Zhou J, Zhao R, Ye T, Yang S, Li Y, Yang F, Wang G, Xie Y, Li Q.
      BACKGROUND AND AIM: Colorectal cancer is one of the most common malignant disease worldwide with highly metastatic potential. Identification of effective therapeutic treatment overcoming such disease is urgent need. Our study focuses on hinokiflavone as an anti-tumor agent against colorectal cancer.METHODS: MTT assay, cell colony formation assay, Hoechst staining, flow cytometer, western blot analysis, real-time PCR, migration and invasion assay were performed to identify effects of hinokiflavone on cell proliferation, apoptosis and metastasis. CT26-tumor bearing mice model was conducted to explore the anti-tumor activity of hinokiflavone in vivo. Immunohistochemistry staining was used to detect the protein expression of Ki-67, cleaved caspase-3 and MMP9 in treated tumors. Acute toxicity was evaluated by serological and hematological analyses and drug side-effects on organs was evaluated by H&E staining.
    RESULTS: Hinokiflavone reduced the proliferation, migration, invasion and promoted the apoptosis in colorectal tumor cells in vitro. Treatment of Hinokiflavone at a tolerable and safe dose (50 mg/kg) significantly suppressed tumor growth in mice bearing CT26 tumors by reducing tumor proliferation, metastasis and inducing apoptosis. Mechanically, treatment of Hinokiflavone induced apoptosis by loss of mitochondrial transmembrane potential and increased reactive oxygen species generation.
    CONCLUSIONS: Hinokiflavone suppressed colorectal tumor cell proliferation, induced apoptosis via the reactive oxygen species-mitochondria-mediated apoptotic pathway, and inhibited tumor cell migration and invasion. Anti-tumor activity of Hinokiflavone was also validated in mice model without observed toxicity. Our findings suggested the plant derived hinokiflavone could be used as an anti-tumor agent against colorectal cancer.
    Keywords:  anti-tumor effect; apoptosis; colorectal cancer; hinokiflavone; metastasis
    DOI:  https://doi.org/10.1111/jgh.14581
  12. Can J Physiol Pharmacol. 2018 Dec 21.
    Cui W, Leng B, Wang G.
      Klotho protein secreted in the blood could act as a hormone to regulate various target organs and had a protective effect on the cardiovascular system. Numerous studies had shown that klotho protein had anti-oxidative stress, anti-inflammatory and anti-apoptotic effects on vascular endothelial cells. The purpose of this study was to investigate the protective mechanism of Klotho protein on oxidative damage of vascular endothelial cells induced by H2O2. Klotho protein significantly enhanced HUVECs viability and increased the activities of antioxidant enzymes (superoxide dismutase, catalase, and heme oxygenase-1 [HO-1]), scavenged reactive oxygen species (ROS), inhibited TNF-α, IL-6 secretion. Klotho protein also reduced the rate of apoptosis of cells and improved the function of vascular endothelial cells (increased NO secretion). Klotho protein activated nuclear translocation of NRF2 and increased HO-1 expression. Klotho protein also activated phosphorylation of AKT, whereas the addition of LY294002, a pharmacological inhibitor of PI3K, blocked KL (klotho protein)-induced Nrf2/HO-1 activation and cytoprotection. Klotho protein enhanced the antioxidant defense ability of the cells by activating the PI3K/AKT pathway, which upregulated the expression of NRF2/HO-1, thereby inhibiting H2O2-induced oxidative damage.
    DOI:  https://doi.org/10.1139/cjpp-2018-0277
  13. Aquat Toxicol. 2018 Dec 06. pii: S0166-445X(18)30676-3. [Epub ahead of print]207 120-131
    Zhao D, Zhang X, Li X, Ru S, Wang Y, Yin J, Liu D.
      Copper (Cu) is one of the most widespread environmental pollutants and is known to exert multiple toxic effects including reproductive toxicity. In this study, we investigated the toxic effect of Cu on reproduction of the red swamp crayfish (Procambarus clarkii), an economic crustacean species, by exposing adult male crayfish to 0.03 and 3.00 mg/L Cu2+ for 7 days. The results showed that Cu2+ exposure induced oxidative stress accompanied by elevated reactive oxygen species (ROS) and malondialdehyde (MDA) levels in testes, and resulted in decreased sperm quality and abnormal testicular structures with apoptotic germ cells and vacuolisation in Sertoli cells. To reveal the molecular mechanism of Cu2+-induced oxidative damage in crayfish testes, we sequenced, assembled and annotated the transcriptome for crayfish testes, using the Illumina sequencing approach. After the 3.00 mg/L Cu2+ treatment, 6745 genes with differentially expressed profile were identified, of which many genes were involved in cellular response to ROS based on Gene Ontology enrichment analysis. Further, KEGG analysis demonstrated that genes with up-regulated expression levels significantly enriched in mitochondria oxidative phosphorylation pathway, suggesting disturbed mitochondrial electron transport chain was probably a main source of Cu2+-induced ROS production in testes. This study represented the first use of transcriptome to investigate the toxic effect of Cu2+ on male crayfish reproduction, and the pathways identified underlying Cu2+ toxicity at molecular level provide a novel insight into the reproductive toxicity of Cu in crustaceans.
    Keywords:  Copper; Male crayfish; Oxidative damage; Reproductive toxicity; Transcriptome
    DOI:  https://doi.org/10.1016/j.aquatox.2018.12.006
  14. Oncol Rep. 2018 Dec 19.
    Kim JL, Lee DH, Jeong S, Kim BR, Na YJ, Park SH, Jo MJ, Jeong YA, Oh SC.
      Imatinib is a powerful tyrosine kinase inhibitor that specifically targets BCR‑ABL, c‑KIT, and PDGFR kinases, and is used in the treatment of chronic myelogenous leukemia, gastrointestinal stromal tumors, and other types of cancers. However, the possible anticancer effects of imatinib in gastric cancer have not yet been explored. The present study evaluated the in vitro effects of imatinib on gastric cancer cells and determined the molecular mechanism underlying these effects. We determined that imatinib induced mitochondria‑mediated apoptosis of gastric cancer cells by involving endoplasmic reticulum (ER) stress‑associated activation of c‑Jun NH2‑terminal kinase (JNK). We also found that imatinib suppressed cell proliferation in a time‑ and dose‑dependent manner. Cell cycle analysis revealed that imatinib‑treated AGS cells were arrested in the G2/M phase of the cell cycle. Moreover, imatinib‑treated cells exhibited increased levels of phosphorylated JNK, and of the transcription factor C/EBP homologous protein, an ER stress‑associated apoptotic molecule. Results of cell viability assays revealed that treatment with a combination of imatinib and chemotherapy agents irinotecan or 5‑Fu synergistically inhibited cell growth, compared with treatment with any of these drugs alone. These data indicated that imatinib exerted cytotoxic effects on gastric cancer cells by inducing apoptosis mediated by reactive oxygen species generation and ER stress‑associated JNK activation. Furthermore, we revealed that imatinib induced the apoptosis of gastric cancer cells by inhibiting platelet‑derived growth factor receptor signaling. Collectively, our results strongly support the use of imatinib in the treatment of treating gastric cancer.
    DOI:  https://doi.org/10.3892/or.2018.6945
  15. Mol Med Rep. 2018 Dec 12.
    Nazim UM, Yin H, Park SY.
      Tumor necrosis factor‑related apoptosis-inducing ligand (TRAIL) is well known as a transmembrane cytokine and has been proposed as one of the most effective anti‑cancer therapeutic agents, owing to its efficiency to selectively induce cell death in a variety of tumor cells. Suppression of autophagy flux has been increasingly acknowledged as an effective and novel therapeutic intervention for cancer. The present study demonstrated that the anti‑cancer and anti‑inflammatory drug celastrol, through its anti‑metastatic properties, may initiate TRAIL‑mediated apoptotic cell death in lung cancer cells. This sensitization was negatively affected by N‑acetyl‑l‑cysteine, which restored the mitochondrial membrane potential (ΔΨm) and inhibited reactive oxygen species (ROS) generation. Notably, treatment with celastrol caused an increase in microtubule‑associated proteins 1A/1B light chain 3B‑II and p62 levels, whereas co‑treatment of celastrol and TRAIL increased active caspase 3 and 8 levels compared with the control, confirming inhibited autophagy flux. The combined use of TRAIL with celastrol may serve as a safe and adequate therapeutic technique for the treatment of TRAIL‑resistant lung cancer, suggesting that celastrol‑mediated autophagy flux inhibition sensitized TRAIL‑initiated apoptosis via regulation of ROS and ΔΨm.
    DOI:  https://doi.org/10.3892/mmr.2018.9757
  16. Drug Chem Toxicol. 2018 Dec 21. 1-7
    Kianfar M, Nezami A, Mehri S, Hosseinzadeh H, Hayes AW, Karimi G.
      Acrylamide (ACR), a vinyl monomer that has multiple chemical and industrial applications, is a neurotoxic agent in human and animal. Fasudil is a potent Rho-kinase inhibitor which exhibits neuroprotective effects in some neuronal degenerative disorders. In this study, the potential protective effect of Fasudil on ACR-induced cytotoxicity in PC12 cells was evaluated. Our results showed that ACR increased the level of intracellular reactive oxygen species (ROS) and consequently upregulated the Bax/Bcl-2 ratio and significantly elevated the level of caspase-3 and 9 proteins in PC12 cells. Interestingly, pretreatment with Fasudil protected PC12 cells against ACR-induced toxicity mainly through the reduction of ROS production and modulation of proteins which involved in apoptosis pathway. Fasudil down-regulated the Bax/Bcl-2 ratio and the levels of caspase-3 and 9 proteins in cells exposed to ACR. In conclusion, the neuroprotective effect of Fasudil against ACR-induced toxicity in PC12 cells appears to be mediated through inhibition of ROS production and modulation of apoptosis.
    Keywords:  Acrylamide; Fasudil; apoptosis; neurotoxicity; rho-kinase inhibitor
    DOI:  https://doi.org/10.1080/01480545.2018.1536140
  17. Bioorg Chem. 2018 Dec 10. pii: S0045-2068(18)30983-0. [Epub ahead of print]84 468-477
    Wang Y, Wu C, Zhang Q, Shan Y, Gu W, Wang S.
      A series of novel β-pinene-based thiazole derivatives were synthesized and characterized by HRMS, 1H NMR, and 13C NMR analyses as potential antineoplastic agents. Derivatives were evaluated for their anticancer activities in vitro, and the data manifested that most target compounds showed potent anti-proliferative activities against three human cancer cell lines. Especially, compound 5g displayed excellent cytotoxic activity against Hela, CT-26, and SMMC-7721 cell lines with IC50 values of 3.48 ± 0.14, 8.84 ± 0.16, and 6.69 ± 0.15 µM, respectively. To determine the underlying mechanism of compound 5g on cell viability, DAPI staining, Annexin-V/PI staining, JC-1 staining, DCFDA staining, and Western blot analysis were performed. Our data showed that compound 5g inhibited cell proliferation by inducing apoptosis and cell cycle arrest of Hela cells at the G0/G1 phase in a dose dependent manner. Further studies revealed that compound 5g enhanced levels of reactive oxygen species (ROS), caused a decrease in mitochondrial membrane potential, increased the release of mitochondrial cytochrome C, and affected the expression of Bax, Bcl-2, caspase-3 and caspase-9. Thus, our findings indicated that compound 5g induced apoptosis in Hela through ROS-mediated mitochondrial dysfunction signaling pathways.
    Keywords:  Anticancer; Mitochondrial membrane potential; Reactive oxygen species; β-Pinene-based thiazole derivatives
    DOI:  https://doi.org/10.1016/j.bioorg.2018.12.010
  18. Int J Mol Med. 2018 Dec 18.
    Xu J, Zhang G, Tong Y, Yuan J, Li Y, Song G.
      Corilagin, a unique component of the tannin family, has been identified in several medicinal plants. In previous literature, corilagin exhibited a marked anticancer property in a variety of human cancer cells. However, the biological effects of corilagin on gastric cancer and the mechanisms involved remain to be fully elucidated. In the present study, it was reported that corilagin induced inhibition of cell growth in SGC7901 and BGC823 cells in a concentration‑dependent manner. It was found that corilagin exhibited less toxicity towards normal GES‑1 cells. Furthermore, the study showed that corilagin induced the apoptosis of gastric cancer cells mainly via activating caspase‑8, ‑9, ‑3 and poly ADP‑ribose polymerase proteins. Simultaneously, it was verified that corilagin triggered autophagy in gastric cancer cells and the inhibition of autophagy improved the activity of corilagin on cell growth suppression. In addition, corilagin significantly increased intracellular reactive oxygen species production, which is important in inhibiting the growth of gastric cancer cells. Finally, it was shown that necroptosis cannot be induced by corilagin‑incubation in SGC7901 and BGC823 cell lines. Consequently, these findings indicate that corilagin may be developed as a potential therapeutic drug for gastric cancer.
    DOI:  https://doi.org/10.3892/ijmm.2018.4031
  19. Med Sci Monit. 2018 Dec 17. 24 9177-9186
    Gao G, Ding H, Zhuang C, Fan W.
      BACKGROUND The purpose of this research was to investigate the effects of hesperidin on hydrogen peroxide (H₂O₂)-induced chondrocytes injury and cartilage degeneration in a rat model of osteoarthritis (OA). MATERIAL AND METHODS Chondrocytes were isolated from rat knee joints and treated with hesperidin alone or combined with H₂O₂. Then, Cell Counting Kit-8 (CCK-8) assay was used to assess cell viability. Activity of reactive oxygen species (ROS) and levels of malondialdehyde (MDA) were estimated. Cell apoptosis was assessed by flow cytometry assay. In addition, gene expression levels were measured for caspase 3, tumor necrosis factor-alpha (TNF-α), interleukin-1beta (IL-1β), collagen type II (Col2a1), aggrecan, (sex-determining region Y)-box 9 (SOX9), matrix metalloproteinase (MMP)-13, and inducible nitric oxide synthase (iNOS) through quantitative real-time polymerase chain reaction (qPCR). To examine the effects on cartilage destruction in vivo, hesperidin or vehicle control were orally administrated in a surgically-induced osteoarthritis (OA) model. RESULTS The results indicated that hesperidin pretreatment of chondrocytes reduce H₂O₂-induced cytotoxicity and apoptosis. Hesperidin pretreatment decreased the formation of MDA and intracellular ROS, including chondrocyte apoptosis. Hesperidin also reversed the activity of H₂O₂ on inhibiting the Col2a1, aggrecan, and SOX9 gene expression and increasing the gene expression of caspase 3, IL-1β, TNFα, iNOS, and MMP13. In addition, hesperidin administration markedly attenuated cartilage destruction and reduced IL-1β and TNF-α levels in a surgically-induced OA model. CONCLUSIONS Our study suggests that hesperidin can prevent H₂O₂-induced chondrocytes injury through its antioxidant effects in vitro and reduce cartilage damage in a rat model of OA.
    DOI:  https://doi.org/10.12659/MSM.913726
  20. PLoS One. 2018 ;13(12): e0209020
    Zhang L, Wu L, Si Y, Shu K.
      The influence of nanomaterials on the ecological environment is becoming an increasingly hot research field, and many researchers are exploring the mechanisms of nanomaterial toxicity on microorganisms. Herein, we studied the effect of two different sizes of nanosilver (10 nm and 50 nm) on the soil nitrogen fixation by the model bacteria Azotobacter vinelandii. Smaller size AgNPs correlated with higher toxicity, which was evident from reduced cell numbers. Flow cytometry analysis further confirmed this finding, which was carried out with the same concentration of 10 mg/L for 12 h, the apoptotic rates were20.23% and 3.14% for 10 nm and 50 nm AgNPs, respectively. Structural damage to cells were obvious under scanning electron microscopy. Nitrogenase activity and gene expression assays revealed that AgNPs could inhibit the nitrogen fixation of A. vinelandii. The presence of AgNPs caused intracellular reactive oxygen species (ROS) production and electron spin resonance further demonstrated that AgNPs generated hydroxyl radicals, and that AgNPs could cause oxidative damage to bacteria. A combination of Ag content distribution assays and transmission electron microscopy indicated that AgNPs were internalized in A. vinelandii cells. Overall, this study suggested that the toxicity of AgNPs was size and concentration dependent, and the mechanism of antibacterial effects was determined to involve damage to cell membranes and production of reactive oxygen species leading to enzyme inactivation, gene down-regulation and death by apoptosis.
    DOI:  https://doi.org/10.1371/journal.pone.0209020
  21. R Soc Open Sci. 2018 Nov;5(11): 180509
    Li Y, Guo M, Lin Z, Zhao M, Xia Y, Wang C, Xu T, Zhu B.
      The morbidity and mortality of hepatocellular carcinoma, the most common cancer, are increasing continuously worldwide. Galangin (Ga) has been demonstrated to possess anti-cancer effect, but the efficacy of Ga was limited by its low permeability and poor solubility. To develop aqueous formulation and improve the anti-cancer activity of Ga, surface decoration of functionalized selenium nanoparticles with Ga (Se@Ga) was synthesized in the present study. The aim of this study was to evaluate the anti-cancer effect of Se@Ga and the mechanism on HepG2 cells. Se@Ga-induced HepG2 cell apoptosis was confirmed by depletion of mitochondrial membrane potential, translocation of phosphatidylserine and caspase-3 activation. Furthermore, Se@Ga enhanced the anti-cancer activity of HepG2 cells through ROS-mediated AKT and p38 signalling pathways. In summary, these results suggest that Se@Ga might be potential candidate chemotherapy for cancer.
    Keywords:  Galangin; apoptosis; caspase-3; reactive oxygen species; selenium nanoparticles
    DOI:  https://doi.org/10.1098/rsos.180509
  22. J Biomed Phys Eng. 2018 Dec;8(4): 447-452
    Arjmandi N, Mortazavi G, Zarei S, Faraz M, Mortazavi SAR.
      Since the early days of human life on the Earth, our skin has been exposed to different levels of light. Recently, due to inevitable consequences of modern life, humans are not exposed to adequate levels of natural light during the day but they are overexposed to relatively high levels of artificial light at night. Skin is a major target of oxidative stress and the link between aging and oxidative stress is well documented. Especially, extrinsic skin aging can be caused by oxidative stress. The widespread use of light emitting diodes (LEDs) and the rapidly increasing use of smartphones, tablets, laptops and desktop computers have led to a significant rise in the exposure of human eyes to short-wavelength visible light. Recent studies show that exposure of human skin cells to light emitted from electronic devices, even for exposures as short as 1 hour, may cause reactive oxygen species (ROS) generation, apoptosis, and necrosis. The biological effects of exposure to short-wavelength visible light in blue region in humans and other living organisms were among our research priorities at the Ionizing and Non-ionizing Radiation Protection Research Center (INIRPRC). Today, there is a growing concern over the safety of the light sources such as LEDs with peak emissions in the blue light range (400-490 nm). Recent studies aimed at investigating the effect of exposure to light emitted from electronic device on human skin cells, shows that even short exposures can increase the generation of reactive oxygen species. However, the biological effects of either long-term or repeated exposures are not fully known, yet. Furthermore, there are reports indicating that frequent exposure to visible light spectrum of the selfie flashes may cause skin damage and accelerated skin ageing. In this paper we have addressed the different aspects of potential effects of exposure to the light emitted from smartphones' digital screens as well as smartphones' photoflashes on premature aging of the human skin. Specifically, the effects of blue light on eyes and skin are discussed. Based on current knowledge, it can be suggested that changing the spectral output of LED-based smartphones' flashes can be introduced as an effective method to reduce the adverse health effects associated with exposure to blue light.
    Keywords:   Blue Light; Mobile Phones ; Selfies ; Skin Aging ; Skin Damage ; Smartphones
  23. Eur J Pharmacol. 2018 Dec 12. pii: S0014-2999(18)30725-8. [Epub ahead of print]
    Xu T, Huang C, Qi XT, Yang XC, Zhang N, Cao J, Wang C, Zhu H, Yang B, He QJ, Shao XJ, Ying MD.
      Osteosarcoma is the most common primary malignant bone tumour, but the survival rate of patients has plateaued since the mid-1980s. Adriamycin is an integral component of the current first-line chemotherapies used for osteosarcoma, but dose-dependent severe side effects often limit its clinical application. Here, we propose a potential combination regimen in which adriamycin plus 2-bromopalmitate, a palmitoylation inhibitor, exhibited powerful therapeutic effects on osteosarcoma. First, 2-bromopalmitate strongly increased the proliferation inhibition of adriamycin in both human osteosarcoma cell lines and primary osteosarcoma cells. Adriamycin-induced apoptosis in osteosarcoma cells was enhanced when synergized with 2-bromopalmitate. Our study indicated that the reactive oxygen species scavenger NAC and GSH could largely reverse the apoptosis induced by adriamycin combined with 2-bromopalmitate, demonstrating that reactive oxygen species played an essential role in this combination therapy. Moreover, CHOP was remarkably elevated in the combination group, and silencing of CHOP almost completely blocked the apoptosis induced by the combination of 2-bromopalmitate and adriamycin. Taken together, our study provides a prospective therapeutic strategy to eliminate osteosarcoma, which is propitious to clinical combination therapy development.
    Keywords:  2-Bromopalmitate; Adriamycin; CHOP; Osteosarcoma; Reactive oxygen species
    DOI:  https://doi.org/10.1016/j.ejphar.2018.12.019
  24. Pharmacol Ther. 2018 Dec 14. pii: S0163-7258(18)30228-6. [Epub ahead of print]
    Mustroph J, Lebek S, Maier LS, Neef S.
      Ethanol can acutely and chronically alter cardiomyocyte and whole-organ function in the heart. Importantly, ethanol acutely and chronically predisposes to arrhythmias, while chronic abuse can induce heart failure. However, the molecular mechanisms of ethanol toxicity in the heart are incompletely understood. In this review, we summarize the current mechanistic knowledge on cardiac ethanol toxicity, with a focus on druggable pathways. Ethanol effects on excitation-contraction coupling, oxidative stress, apoptosis, and cardiac metabolism, as well as effects of ethanol metabolites will be discussed. Important recent findings have been gained by investigation of acute ethanol effects. These include a renewed focus reactive oxygen species (ROS) and induction of SR Ca2+ leak by CaMKII-mediated pathways downstream of ROS. Furthermore, a clinical outlook into potential novel treatment options is provided.
    Keywords:  Alcohol; Arrhythmogenesis; CaMKII; Cardiomyocytes; Ethanol; Heart
    DOI:  https://doi.org/10.1016/j.pharmthera.2018.12.006
  25. Food Nutr Res. 2018 ;62
    Zhang X, Song Z, You Y, Li X, Chen T.
      Background: Phoenix Dan Cong tea is an Oolong tea produced in Chaozhou, China. Nowaday, the experimental studies on the benefical effects of the Phoenix Dan Cong tea are rare.Objective: The objective of this study was to comprehensively evaluate the activity of Phoenix Dan Cong tea aqueous extract (PDCe).
    Methods: We used a series of evaluation methods in the present study to achieve an in-depth understanding and evaluation of the antioxidant and antitumor activity of PDCe.
    Results: High-performance liquid chromatography (HPLC) studies have indicated that PDCe is rich in catechins such as gallocatechin (GC), epigallocatechin (EGCG) and epicatechin gallate (ECG), with sparse amounts of theaflavins. We discovered that PDCe scavenges ABTS•+ and DPPH• free radicals in a dose-dependent manner. In addition, PDCe can significantly induce apoptosis of MDA-MB231 cells, mainly through the death-receptor-mediated extrinsic apoptotic pathway. Internalized PDCe can not only downregulate intracellular reactive oxygen species levels but also induce oxidative damage to mitochondria in MDA-MB231 cells.
    Conclusions: Phoenix Dan Cong tea may act as a substitute for natural antioxidants and as a promising anticancer agent due to its protective effect on human health.
    Keywords:  Dan Cong tea; aqueous extract; cell apoptosis; free radical scavenging; oxidative damage; protective effect
    DOI:  https://doi.org/10.29219/fnr.v62.1500
  26. Eur Rev Med Pharmacol Sci. 2018 Dec;pii: 16651. [Epub ahead of print]22(24): 8830-8838
    Qu XH, Zhang K.
      OBJECTIVE: Phosphatidylinositol 3-kinase/protein kinase B ((PI3K/AKT) signaling pathway plays a role in regulating cell survival and apoptosis. Phosphatase and tensin homologue deleted on chromosome ten (PTEN) can negatively regulate PI3K/AKT signaling pathway, while DJ-1 (Parkinson gene 7) can negatively regulate expression and function of PTEN. DJ-1-PTEN/PI3K/AKT signaling pathway plays a role in the regulation of ischemic reperfusion (IR) injury. Bioinformatics analysis showed that there was a targeted complementary binding site between microRNA-122 (miR-122) and 3'-UTR of DJ-1 mRNA. This study aimed to investigate the effects of miR-122 in regulating DJ-1-PTEN/PI3K/AKT signaling pathway and acute renal I-R injury.MATERIALS AND METHODS: Rat renal artery was clamped and restored after 30 min to establish renal IR injury model. Renal tissue samples were collected at 10 h and 20 h after operation. miR-122 and DJ-1 mRNA were detected with quantitative Real-time PCR (qRT-PCR). DJ-1 protein was tested by using Western blot. Blood urea nitrogen (BUN) and serum creatinine (SCr) were measured. Rat tubular epithelial cells, RRTEC, were cultured in vitro and divided into transfection (miR-NC group) and treatment group (miR-122 inhibitor group). IR treatment was performed after 72 h of transfection. DJ-1, PTEN, AKT, and phosphorylated AKT (p-AKT) were detected using Western blot. Cell apoptosis and reactive oxygen species (ROS) were measured with flow cytometry.
    RESULTS: Compared with Sham group, blood BUN and SCr contents significantly increased (p < 0.05), miR-122 level significantly elevated (p < 0.05), while DJ-1 mRNA and protein markedly declined (p < 0.05) in IR model rats. Compared with control group, I-R treatment significantly up-regulated miR-122 and PTEN expressions (p < 0.05), decreased DJ-1 and p-AKT levels (p < 0.05), and induced apoptosis and ROS production (p < 0.05) in RRTEC cells. Transfection with miR-122 inhibitor markedly up-regulated DJ-1 expression (p < 0.05), enhanced PTEN/PI3K/AKT pathway activity (p < 0.05), and reduced apoptosis and ROS production (p < 0.05).
    CONCLUSIONS: MiR-122 increased significantly, while DJ-1 declined significantly during renal I-R injury. Down-regulation of miR-122 markedly elevated DJ-1, enhanced PTEN/PI3K/AKT pathway activity, and inhibited apoptosis and ROS generation in rat renal tubular epithelial cells to alleviate IR injury.
    DOI:  https://doi.org/10.26355/eurrev_201812_16651
  27. Cell Biol Int. 2018 Dec 20.
    Cai HA, Tao X, Zheng LJ, Huang L, Peng Y, Liao RY, Zhu YM.
      Cerebral ischemia reperfusion (I/R) injury is commonly observed, and often causes severe complications. Ozone has been applied for protecting ischemia/reperfusion (I/R) injury in animal models of several organs including cerebra, but the detailed mechanism remains unclear. MTT assay and LDH measurement were used to determine the influence of ozone on cell activity and damage of SH-SY5Y cells. Some redox items such as CAT, MDA, GSH-Px and SOD were measured by ELISA assay. The mitochondrial membrane potential (ΔΨm) was determined by JC-1 assay. Cytochrome-c (cyt-c) level in cytoplasm and mitochondrion was measured by western blotting. Apoptosis was determined by flow cytometry, and some apoptosis-related molecules were detected by qRT-PCR and western blotting. Ozone alleviated oxidative damage by increasing GSH-Px, SOD, CAT, and decreasing MDA. Ozone decreased mitochondrial damage caused by I/R injury and inhibited the release of cyt-c from mitochondrion to cytoplasm in SH-SY5Y cells. Cell apoptosis caused by I/R was inhibited by ozone, and ozone could decrease apoptosis by increasing ratio of Bcl-2/Bax and inhibiting caspase signaling pathway in SH-SY5Y cells. Ozone has the ability of maintaining redox homeostasis, decreasing mitochondrion damage, inhibiting neurocytes apoptosis induced by I/R. Therefore, ozone may be a promising protective strategy against cerebral I/R injury.
    Keywords:  Cell apoptosis; Cerebral ischemia reperfusion; Mitochondrial damage; Ozone
    DOI:  https://doi.org/10.1002/cbin.11087
  28. Int J Mol Sci. 2018 Dec 14. pii: E4050. [Epub ahead of print]19(12):
    Wang YQ, Wei JG, Tu MJ, Gu JG, Zhang W.
      Acetaminophen (APAP) is a widely used analgesic and antipyretic drug that leads to severe hepatotoxicity at excessive doses. Fucoidan, a sulfated polysaccharide derived from brown seaweeds, possesses a wide range of pharmacological properties. However, the impacts of fucoidan on APAP-induced liver injury have not been sufficiently addressed. In the present study, male Institute of Cancer Research (ICR) mice aged 6 weeks were subjected to a single APAP (500 mg/kg) intraperitoneal injection after 7 days of fucoidan (100 or 200 mg/kg/day) or bicyclol intragastric administration. The mice continued to be administered fucoidan or bicyclol once per day, and were sacrificed at an indicated time. The indexes evaluated included liver pathological changes, levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) in the serum, levels of reactive oxygen species (ROS), malondialdehyde (MDA), superoxide dismutase (SOD), glutathione (GSH) and catalase (CAT) in the liver, and related proteins levels (CYP2E1, pJNK and Bax). Furthermore, human hepatocyte HL-7702 cell line was used to elucidate the potential molecular mechanism of fucoidan. The mitochondrial membrane potential (MMP) and nuclear factor-erythroid 2-related factor (Nrf2) translocation in HL-7702 cells were determined. The results showed that fucoidan pretreatment reduced the levels of ALT, AST, ROS, and MDA, while it enhanced the levels of GSH, SOD, and CAT activities. Additionally, oxidative stress-induced phosphorylated c-Jun N-terminal protein kinase (JNK) and decreased MMP were attenuated by fucoidan. Although the nuclear Nrf2 was induced after APAP incubation, fucoidan further enhanced Nrf2 in cell nuclei and total expression of Nrf2. These results indicated that fucoidan ameliorated APAP hepatotoxicity, and the mechanism might be related to Nrf2-mediated oxidative stress.
    Keywords:  Nrf2; acetaminophen; fucoidan; hepatotoxicity; oxidative stress
    DOI:  https://doi.org/10.3390/ijms19124050
  29. Mol Med Rep. 2018 Dec 11.
    Wang Q, Yu H, Yu H, Ma M, Ma Y, Li R.
      Dexmedetomidine (DEX), an α2 adrenoceptor agonist, is a commonly used anesthetic drug in surgical procedures. Previous studies have indicated that DEX exerts neuroprotective effects. However, the molecular mechanism underlying this process remains to be elucidated. The present study investigated a potential implication of microRNA (miR)‑223‑3p in the DEX‑induced anti‑oxidative effect on neuronal cells. The results indicated that following hydrogen peroxide (H2O2)‑mediated induction of oxidative stress, the viability of human hippocampal neuronal cells was markedly decreased, as determined by an MTT assay. In addition, treatment with H2O2 induced cell apoptosis, the release of lactate dehydrogenase, accumulation of intracellular calcium, phosphorylation of calmodulin‑2, and production of malondialdehyde and reactive oxygen species. Furthermore, treatment with H2O2 inhibited the expression of mir‑223‑3p and enhanced the expression of its target cytotoxic granule associated RNA binding protein like 1 (TIAL1), and these effects were reversed by treatment with DEX. Mechanistic studies demonstrated that the 3'‑untranslated region of TIAL1 is a direct target of mir‑223‑3p. The results of the present study demonstrated that DEX may induce its neuroprotective effects by regulating the interaction between miR‑223‑3p and TIAL1. Therefore, the manipulation of miR‑223‑3p/TIAL1 interaction may be involved in the neuroprotective effects of DEX.
    DOI:  https://doi.org/10.3892/mmr.2018.9742
  30. Invest Ophthalmol Vis Sci. 2018 Dec 03. 59(15): 6014-6025
    Tang W, Ma J, Gu R, Lei B, Ding X, Xu G.
      Purpose: Lipocalin 2 (LCN2) is reported to be one of the key regulators of cell survival and death; however, its effect on retinal degeneration is unclear. Therefore, we aimed to investigate the role of LCN2 and its underlying mechanisms in light-induced retinal degeneration.Methods: A recombinant lentivirus expressing a short hairpin RNA targeting LCN2 mRNA and a recombinant lentivirus overexpressing LCN2 were used to downregulate and upregulate retinal LCN2, respectively. Seven days after intravitreal injection of the lentiviruses, rats were exposed to blue light (2500 lux) for 24 hours. Retinal function and morphology were evaluated with ERG and hematoxylin-eosin staining, respectively. TUNEL staining was used to detect apoptotic cells. The levels of reactive oxygen species (ROS) were evaluated with dihydroethidium labeling. Western blotting and real-time PCR were used to examine protein and mRNA expression levels, respectively.
    Results: Retinal LCN2 expression was significantly upregulated after light exposure. Light exposure reduced the amplitudes of a- and b-waves on the ERG and the thickness of the outer nuclear layer and promoted photoreceptor apoptosis. These phenomena were clearly attenuated by LCN2 knockdown, whereas LCN2 overexpression had the opposite effects. The overexpression of LCN2 facilitated photoreceptor apoptosis by increasing ROS generation and Bim expression. On the opposite, LCN2 knockdown mitigated the generation of light-exposure-induced ROS and the activation of the Bim-mediated mitochondrial apoptotic pathway.
    Conclusions: Light-induced LCN2 is a proapoptotic factor in the retina, and LCN2 knockdown protects photoreceptors from apoptosis by inhibiting ROS production and Bim expression. LCN2 is a potential therapeutic target for light-induced retinal degeneration.
    DOI:  https://doi.org/10.1167/iovs.18-25213
  31. Front Neurosci. 2018 ;12 868
    Li M, Zhu Y, Peng W, Wang H, Yuan Y, Gu X.
      ABPPk, the active ingredient separated from Achyranthes bidentata polypeptides, is a traditional Chinese medicine with multiple pharmaceutical properties. In this study, we investigated the molecular mechanisms of ABPPk in protecting Schwann cells (SCs) from H2O2-induced cell apoptosis. The viability of SCs pretreated with ABPPk was elevated significantly by MTT assay estimation. Meanwhile, the apoptosis of SCs was reduced which was showed in flow cytometry and transferase-mediated dUTP nick end labeling analysis. Furthermore, the addition of ABPPk also increased the activities of SOD and GSH accompanied with a decrease in MDA and LDH activities. According to Western blot analysis, the upregulation of Bcl-2, also downregulation of Bax and cleaved caspase-3 were demonstrated in SCs which was ABPPk pretreated. Further research showed that PI3K/AKT and ERK1/2 pathways in SCs have been activated after pretreatment of ABPPk. Collectively, results in our study suggested that ABPPk protected SCs from H2O2-induced oxidative damage by reducing the expression of apoptotic molecules and enhancing the activities of antioxidant enzymes, which inhibited the apoptosis of SCs modulated by PI3K/AKT and ERK1/2 signaling pathways. In our perspectives, ABPPk as an active factor with its antioxidative activities has potential and promising therapeutic effects in the prevention of neurologic disorders.
    Keywords:  Achyranthes bidentata polypeptides; cell apoptosis; hydrogen peroxide; oxidative stress; schwann cells
    DOI:  https://doi.org/10.3389/fnins.2018.00868
  32. Mol Med Rep. 2018 Dec 18.
    Kim JS, Shin SJ, Kim JN, Kwon MJ, Lim EY, Kim YT, Kim H, Kim BJ.
      Traditional herbal medicines are being increasingly used worldwide to treat cancer. Radix Sophorae Flavescentis (RSF) is a Chinese herb, which has numerous pharmacological properties, including anti‑tumour effects. In this study, we investigated the mechanisms underlying RSF‑induced apoptosis in human gastric cancer cells (AGS cells). We found that RSF treatment (20‑200 µg/ml) inhibited the proliferation of AGS cells and increased the sub‑G1 phase ratio. RSF‑induced cell death was associated with the downregulation of BCl‑2 and upregulation of Bax. In addition to increasing the expression levels of apoptosis‑mediating surface antigen FAS and Fas ligand, RSF also activated caspase‑3; however, mitogen‑activated protein kinase appeared to inhibit RSF‑induced cell death. RSF also led to an increased production of reactive oxygen species. Based on these results, we propose that RSF could be a potential therapeutic agent for gastric cancer.
    DOI:  https://doi.org/10.3892/mmr.2018.9776
  33. Acta Biochim Biophys Sin (Shanghai). 2018 Dec 19.
    Liu F, Yang L, Zheng Y, Zhang W, Zhi J.
      Preeclampsia not only seriously endangers maternal and fetal health during pregnancy but may incur many sequelae in postpartum women such as reduced visual acuity. Agonistic autoantibodies to the angiotensin II type I receptor (AT1-AA) is closely associated with preeclampsia. The aim of the present study is to determine whether AT1-AA is associated with retinal impairment during the course of preeclampsia. A preeclampsia model was established by injecting AT1-AA into pregnant rats via the tail vein. Changes in the retinal histological structure were observed. Cell apoptosis and cytokines including reactive oxygen species (ROS), as well as apoptosis-related proteins such as Bcl-2, Bax, and caspase-3 were detected. In addition, flash electroretinograms obtained at different postpartum days were analyzed. Compared with the control group, the retinal structure became edematous and the cell density was reduced significantly in preeclampsia group. The cell apoptosis rate was increased significantly. In addition, the content of ROS, the levels of Bax and caspase-3 in the retina were increased, while the content of Bcl-2 was reduced significantly. Continuous observation of the electroretinograms showed loss of retinal ganglion cells postpartum. The present study demonstrated that AT1-AA induced retinal cell apoptosis by promoting ROS release and activating caspase, suggesting that the increased postpartum susceptibility of preeclamptic women to retinopathy is related to AT1-AA-induced cell apoptosis.
    DOI:  https://doi.org/10.1093/abbs/gmy144
  34. J Cancer Res Clin Oncol. 2018 Dec 19.
    Chen L, Yang H, Yi Z, Jiang L, Li Y, Han Q, Yang Y, Zhang Q, Yang Z, Kuang Y, Zhu Y.
      PURPOSE: Clinical outcomes for advanced malignant melanoma (MM) are often poor due to tumor invasiveness, metastasis, recurrence, and multidrug resistance.METHODS: We investigated whether apoptosis, cell cycle regulation, oxidative status, and redox balance were altered by changes in the expression of the long noncoding RNA, growth arrest-specific transcript 5 (GAS5), in MM cells.
    RESULTS: Analysis of clinical samples from MM patients showed that the rate of reduced GAS5 expression, relative to that in adjacent noncancerous tissues, was significantly lower for tumors from patients with advanced disease (76.6%, P < 0.001), as evidenced by larger tumor size, higher TNM stage, and higher incidences of ulceration and metastasis (P < 0.001 for all). Cell culture experiments showed that siRNA-mediated knockdown of GAS5 increased the viability of A375-GAS5si cells. Flow cytometry and western blotting showed that GAS5 knockdown increased MM cell proliferation by inducing G1/S cell cycle progression through increases in Cyclin D1, CDK4, and p27 expression (P < 0.05 for all) and by inhibiting apoptosis through an increase in Bcl-2 expression (P < 0.001). Knockdown of GAS5 also increased levels of superoxide anion (P < 0.01), NADP+(P < 0.001), and oxidized glutathiones (P < 0.01) through increases in NOX4 expression (P < 0.001), G6PD expression (P < 0.01), and NOX activity (P < 0.05), and RNA co-immunoprecipitation showed that GAS5 induced these changes through a physical interaction between GAS5 and the G6PD protein.
    CONCLUSIONS: Our findings show GAS5 contributes to regulation of the apoptosis, cell cycle, homeostasis of reactive oxygen species, and redox balance in MM cells, and suggest that reduced GAS5 expression contributes to disease progression in MM patients.
    Keywords:  Apoptosis; Cell cycle; GAS5; LncRNA; Malignant melanoma; Reactive oxygen species
    DOI:  https://doi.org/10.1007/s00432-018-2820-4
  35. Int J Mol Med. 2018 Dec 18.
    Wang C, Cai X, Hu W, Li Z, Kong F, Chen X, Wang D.
      Due to its complex pathogenesis, the prevention and therapization of Alzheimer's disease (AD) remains a serious challenge. Crocin, the main compound isolated from Crocus sativus L., demonstrates various pharmacological activities including anti‑apoptotic properties. The present study investigated the neuroprotective effect of crocin and the underlying mechanisms. In l‑glutamate‑damaged HT22 cells, 3‑h crocin pretreatment strongly enhanced the HT22 cell viability, reduced the apoptotic rate, mitigated mitochondrial dysfunction, suppressed intracellular reactive oxygen species (ROS) accumulation and Ca2+ overload compared with untreated cells. Additionally, crocin significantly decreased the expression levels of Bax, Bad and cleaved caspase‑3 and increased the expression levels of B‑cell lymphoma‑extra large, phosphorylated (P‑) protein kinase B and P‑mammalian target of rapamycin compared with untreated cells. In mice with AD induced by d‑galactose and aluminum trichloride, crocin substantially improved the cognition and memory abilities of the mice as measured by their coordination of movement in an open field test, and reduced their escape time in the Morris water maze test compared with untreated mice. Biochemical analysis confirmed that crocin was able to reduce the Aβ1‑42 content in the mouse brains, increase the levels of glutathione peroxidase, superoxide dismutase, acetylcholine and choline acetyltransferase, and reduce the levels of ROS and acetylcholinesterase in the serum, cerebral cortex and hypothalamus compared with untreated mice. Immunohistochemical analysis demonstrated that crocin reduced Aβ1‑42 deposition in the hippocampus of the brains of treated mice compared with untreated mice. In conclusion, crocin demonstrates good prospects in the treatment of AD through the oxidative stress‑associated apoptosis signaling pathway.
    DOI:  https://doi.org/10.3892/ijmm.2018.4032
  36. Front Aging Neurosci. 2018 ;10 389
    Liu Y, Li R, Xie J, Hu J, Huang X, Ren F, Li L.
      Oxidative stress is one of the main causes of AMD. Hydrogen has anti-oxidative stress and apoptotic effects on retinal injury. However, the effect of hydrogen on AMD is not clear. In this study, fundus radiography, OCT, and FFA demonstrated that HRW reduced the deposition of drusen-like structures in RPE layer, prevented retina from thinning and leakage of ocular fundus vasculature induced by NaIO3. ERG analysis confirmed that HRW effectively reversed the decrease of a-wave and b-wave amplitude in NaIO3-mice. Mechanistically, HRW greatly reduced the oxidative stress reaction through decreased MDA levels, increased SOD production, and decreased ROS content. The OGG1 expression was downregulated which is a marker of oxidative stress. Involvement of oxidative stress was confirmed using oxidative stress inhibitor ALCAR. Moreover, oxidative stress reaction was associated with expression of Sirt1 level and HRW significantly inhibited the downregulation of Sirt1 expression. This result was further confirmed with AICAR which restore Sirt1 expression and activity. In addition, NaIO3-induced retinal damage was related to apoptosis via caspase 8 and caspase 9, but not the caspase 3 pathways, which led to upregulation of Bax and p53, downregulation of Bcl-2, and increase in Jc-1-positive cells in mice. However, HRW effectively reversed these effects that apoptosis induced. These results suggest that HRW protects retinal functions against oxidative stress injury through inhibiting downregulation of Sirt1 and reducing retinal apoptosis. Therefore, we speculated that hydrogen administration is a promising treatment for AMD therapy.
    Keywords:  AMD; apoptosis; hydrogen; oxidative stress; sirt1
    DOI:  https://doi.org/10.3389/fnagi.2018.00389
  37. Am J Cancer Res. 2018 ;8(11): 2267-2283
    Rabi T, Li F.
      Activating mutations in GTPase protein KRAS occurs in approximately 90% of pancreatic cancers. Mutated KRAS lead to constitutive activation of RAF/MEK/ERK and PI3K/Akt pathways in pancreatic cancer. There is currently no effective KRAS-targeted therapeutics available in the clinic for treating this subset of cancer. In this study we demonstrate that combination of a plant-isolated triterpenoid compound AMR-MeOAc with a low concentration of an antiapoptotic protein inhibitor, FL118 exhibited synergistic cytotoxic activity against pancreatic cancer cells with either mutant KRAS (HPAF-II, KRASG12D) or wild type KRAS (BxPC-3, KRASWT). In pancreatic cancer cells with mutant KRASG12D, AMR-MeOAc and FL118 acting together to inhibit the constitutive KRASG12D mutant activity, increase the reactive oxygen species (ROS) formation, apoptosis induction, and decrease of the expression of survivin and XIAP, while strongly inducing Bax. These effects were also associated with the decrease of B-RAF, ERK and p-ERK. Additionally, AMR-MeOAc and FL118 alone or in combination inhibited the constitutive activation of NF-κB in BxPC-3 cells, which suggests that inhibition of NF-κB in BxPC-3 cells by AMR-MeOAc and FL118 may also be a part of the mechanism of action, when pancreatic cancer cells possess wild type KRAS. Together, the novel combination treatment might provide an effective strategy to overcome the KRASG12D mutant-mediated and NF-κB activation-mediated resistance in pancreatic cancer with either KRASG12D mutation or NF-κB activation/wild type KRAS.
    Keywords:  AMR-MeOAc; ERK; FL118; KRASG12D; NF-κB; XIAP; pancreatic cancer; reactive oxygen species (ROS); survivin
  38. Photodiagnosis Photodyn Ther. 2018 Dec 15. pii: S1572-1000(18)30307-7. [Epub ahead of print]
    Huang L, Chen Q, Yu L, Bai D.
      BACKGROUND: This study aimed to determine the effect of pyropheophorbide-α methyl ester (MPPa)-mediated photodynamic therapy (MPPa-PDT) on the apoptosis and inflammation of murine macrophage RAW264.7 cells.METHODS: Uptake and subcellular localization of MPPa was detected by flow cytometry and confocal fluorescence microscope. Cell viability was assessed by CCK-8; ROS levels were assessed by DCFH-DA. Cell apoptosis was measured by flow cytometry and Hoechst 33342 staining, whereas mitochondrial membrane potential was detected by JC-1 staining. Secretion of tumor necrosis factor α (TNF-α), interleukin-1β (IL-1β), and interleukin-6 (IL-6) was determined using ELISA kits. Caspase-3, cleaved caspase-3, procaspase-9, cleaved caspase-9, PARP, cleaved PARP, Bcl-2, Bax, NF-κB p-p65, p-IKKα/β, and p-IκBα were measured by western blotting. Nuclear factor κB (NF-κB)-p65 nuclear translocation was observed by immunofluorescence.
    RESULTS: MPPa -PDT influenced cell viability in a light dose-dependent manner. It induced ROS formation and RAW264.7 cell apoptosis. It also increased the expression of cleaved caspase-3, cleaved caspase-9, cleaved PARP and Bax, decreased the expression of Bcl-2. While TNF-α, IL-1β, and IL-6 increased in LPS group (model of inflammation), it deceased in LPS-MPPa-PDT group. NF-κB p-p65, p-IKKα/β, and p-IκBα had higher expression in LPS group while that reduced in LPS-MPPa-PDT group. Simultaneously, MPPa-PDT inhibited nuclear translocation of NF-κB-p65 caused by LPS.
    CONCLUSIONS: MPPa-PDT can induce apoptosis and attenuate inflammation in mouse RAW264.7 macrophages, thereby suggesting a promising therapy for atherosclerosis.
    Keywords:  Apoptosis; Atherosclerosis; Inflammation; MPPa-PDT; RAW264.7
    DOI:  https://doi.org/10.1016/j.pdpdt.2018.12.002
  39. Oncol Rep. 2018 Dec 19.
    Kawala RA, Ramadhani FJ, Choi HJ, Lee EH, Park CH, Chung BY, Bai HW.
      Kenalog is a synthetic glucocorticoid drug used to treat various cancers including ocular and choroidal melanoma. However, the drug achieves rarely sustainable results for patients. To overcome this difficulty, the structure of Kenalog was altered by ionizing radiation (IR) to develop a more effective anticancer agent for treatment of various skin cancers. The anticancer effect of modified Kenalog (Kenalog‑IR) was assessed in melanoma cancer cells in vitro. The assessment of mitochondrial functions by MTT assay revealed significant inhibition of melanoma cancer cell viability by Kenalog‑IR compared to Kenalog. Moreover, Kenalog‑IR‑induced apoptotic cell death was associated with the intrinsic mitochondrial pathway by triggering the release of intrinsic apoptosis molecules through activation of caspase‑related molecules in concentration and time‑dependent manners. Furthermore, it was observed that Kenalog‑IR‑induced apoptosis was associated with the generation of reactive oxygen species (ROS) with increased G2/M cell cycle arrest. Collectively, Kenalog‑IR may be a potential suppressor of skin‑related cancer in particular melanoma cancer.
    DOI:  https://doi.org/10.3892/or.2018.6940
  40. Food Chem Toxicol. 2018 Dec 15. pii: S0278-6915(18)30892-5. [Epub ahead of print]
    Štern A, Rotter A, Novak M, Filipič M, Žegura B.
      The cyanobacterial pentapeptide nodularin (NOD), mainly produced by genus Nodularia, is a potent inhibitor of protein phosphatases PPA1 and PPA2, and causes animal mortality. The few studies available indicate that NOD is a potential non-genotoxic carcinogen. In the present study we evaluated NOD (0.01, 0.1 and 1 μg/ml) genotoxic activity in human hepatoma (HepG2) cells with the comet, γH2AX and cytokinesis block micronucleus cytome assays. In addition, induction of oxidative stress was studied. Moreover changes in the expression of selected genes from the P53 pathway, involved in the response to DNA damage (P53, GADD45α, CDKN1A, MDM2), apoptosis (BAX, BCL2) and oxidative stress (GPX1, GSR, GCLC, CAT, SOD1) were determined using qPCR. Non-cytotoxic concentrations induced time and dose dependant increase in reactive oxygen species (ROS) production and substantially increased the formation of oxidative DNA damage. In addition, elevated formation of micronuclei was detected. For the first time it has been shown that NOD deregulated the mRNA level of DNA damage (CDKN1A, GADD45α) and oxidative stress (GPX1, GSR, GCLC, CAT and SOD1) responsive genes and anti-apoptotic gene BCL2. Our results provide new evidence that NOD genotoxic effects are mediated through ROS production, already at low environmentally relevant concentrations.
    Keywords:  Gene expression; HepG2 cells; Micronuclei; Nodularin; Oxidative DNA damage
    DOI:  https://doi.org/10.1016/j.fct.2018.12.019