bims-aporos Biomed news
on Apoptosis and reactive oxygen species
Issue of 2019‒02‒24
fifty-four papers selected by
Gavin McStay
Staffordshire University

  1. J Physiol Biochem. 2019 Feb 22.
    Hosseinzadeh A, Bahrampour Juybari K, Kamarul T, Sharifi AM.
      The high glucose concentration is able to disturb chondrocyte homeostasis and contribute to OA pathogenesis. This study was designed to investigate the protective effects of atorvastatin (ATO) on high glucose (HG)-mediated oxidative stress and mitochondrial apoptosis in C28I2 human chondrocytes. The protective effect of ATO (0.01 and 0.1 μM) on HG (75 mM)-induced oxidative stress and apoptosis was evaluated in C28I2 cells. The effects of ATO on HG-induced intracellular ROS production and lipid peroxidation were detected and the protein expression levels of Bax, Bcl-2, caspase-3, total and phosphorylated JNK and P38 MAPKs were analyzed by Western blotting. The mRNA expression levels of antioxidant enzymes including heme oxygenase-1, NAD(P)H quinine oxidoreductase, glutathione S-transferase-P1, catalase, superoxide dismutase-1, glutathione peroxidase-1, -3, -4 were evaluated by reverse transcription-polymerase chain reaction. Pretreatment with ATO remarkably increased the gene expression levels of antioxidant enzymes and reduced HG-induced elevation of ROS, lipid peroxidation, Bax/Bcl-2 ratio, caspase-3 activation, and JNK and P38 phosphorylation. Atorvastatin could considerably reduce HG-induced oxidative stress and mitochondrial apoptosis through increasing the expression of antioxidant enzymes. Atorvastatin may be considered as a promising agent to prevent high glucose-induced cartilage degradation in OA patients.
    Keywords:  Apoptosis; Atorvastatin; High glucose; Osteoarthritis; Reactive oxygen species
  2. J Cell Biochem. 2019 Feb 21.
    Wang M, Xu T.
      Alzheimer's disease (AD) is the most common aging-associated dementia. The population of AD patients is increasing as the world age grows. Currently, there is no cure for AD. Given that methyl vitamin B12 (methylcobalamin) deficiency is related to AD and Aβ-induced oxidative damage and that methylcobalamin can scavenge reactive oxygen species (ROS) by direct or indirect ways, we studied the effect of methylcobalamin on the cytotoxicity of Aβ. PC12 cells were chronically exposed (24 hours) to Aβ25-35 (25 μM) to establish an AD cell model. The cells were pretreated with or without methylcobalamin (1-100 μM) to investigate the role of methylcobalamin. Cell viability and apoptosis were tested, followed by testing of mitochondrial damage, oxidative stress, and mitochondrial calcium concentration. We observed that methylcobalamin improved the cell viability by decreasing the ratio of apoptosis cells in this AD cell model. Further experiments suggested that methylcobalamin functioned as an antioxidant to scavenge ROS, reducing the endoplasmic reticulum-mitochondria calcium flux through IP3R, preventing mitochondria dysfunction, ultimately protecting cells against apoptosis and cell death. Taken together, our results presented, for the first time, that methyl vitamin B12 can protect cells from Aβ-induced cytotoxicity and the mechanism was mainly relevant to the antioxidative function of methyl B12.
    Keywords:  Alzheimer's disease; Aβ25-35; mitochondria; reactive oxygen species (ROS); vitamin B12
  3. Folia Neuropathol. 2018 ;pii: 34471. [Epub ahead of print]56(4): 354-361
    Rajabian A, Sadeghnia HR, Moradzadeh M, Hosseini A.
      Glutamate is considered to be responsible for the pathogenesis of many neurodegenerative diseases. Reactive oxygen species (ROS) production is considered to be involved in the glutamate-induced apoptosis process. In this study, we investigated the neuroprotective effects of Rheum turkestanicum in the glutamate-induced rat pheochromocytoma (PC12 cells) and mouse neuroblastoma (N2a) cell lines. Rutin as an antioxidant was used as positive control. Glutamate cytotoxicity was accompanied by an increment of malondialdehyde (MDA) content, ROS generation and apoptosis induction. However, pretreatment with the root extract of R. turkestanicum significantly reduced MDA content, ROS generation and apoptotic cell death. Also rutin at a dose of 100 µM reduced ROS production and protected against glutamate toxicity. Also the quantification of rutin in R. turkestanicum extract was achieved and was about 0.11% ± 0.01 w/w. All these findings indicated that R. turkestanicum protected PC12 and N2a cells against glutamate-induced oxidative cell death and apoptosis and might raise the possibility of R. turkestanicum usage as a neuroprotective agent.
    Keywords:   N2a; PC12; apoptosis; glutamate; oxidative damage; rutin; Rheum turkestanicum
  4. Int J Biol Macromol. 2019 Feb 15. pii: S0141-8130(18)36255-X. [Epub ahead of print]
    Li H, Liu D, Li S, Xue C.
      A 1,6-α-D-glucan (CPA) with a high molecular weight of 2.0 × 103 kDa was isolated from the fruits of Castanea mollissima Blume. CPA showed good water solubility, low side effects on normal cells, and moderate cytotoxicity on cancer cells. Selenium nanoparticles (SeNPs) possess significant anticancer effects, but poor water solubility and dispersity. Therefore, an effective nanocomposite was designed in order to enhance the stability and anticancer effects of SeNPs decorated with CPA. Water soluble and dispersed selenium nanoparticles (CPA-SeNPs) were fabricated using CPA and Na2SeO3 by redox reaction with ascorbic acid. The average diameter of CPA-SeNPs was 53.7 ± 4.0 with a Se content of approximately 171.4 mg/g. CPA-SeNPs showed a significant anti-proliferative effect on HeLa cells in vitro. Since these NPs could induce apoptosis and S phase arrest. The apoptotic pathway triggered by CPA-SeNPs on HeLa cells was also characterized as induced by reactive oxygen species (ROS) overproduction, mitochondrial dysfunction and activation of caspase-3. This study suggested that CPA-SeNPs could be developed as a potential candidate in cancer treatment, especially against human cervical cancer.
    Keywords:  Anti-proliferative effect; Apoptosis; Castanea mollissima Blume; Cervical cancer; Glucan; Selenium nanoparticles
  5. Gynecol Endocrinol. 2019 Feb 20. 1-5
    Teixeira CP, Florencio-Silva R, Sasso GRS, Carbonel AAF, Simões RS, Simões MJ.
      Ovarian aging is characterized by declines in follicular reserve and oocyte quality due, in part, to increased oxidative stress and apoptosis. Soy isoflavones (ISOs) have been shown to improve ovarian lifespan by acting as antioxidant and antiapoptotic agents. We aimed at evaluating whether ISOs could modulate oxidative stress and reduce apoptosis and improve ovarian follicle survival in middle-aged female rats. Twelve ovary-intact female Wistar rats (12-month-old) were divided into groups: control (CTRL) and ISO, daily treated by gavage with vehicle or soy-ISO extract (150 mg/kg b.w), respectively. After 8 weeks, rats were euthanized and their ovaries removed for histomorphometric (% follicles) and apoptosis (cleaved-caspase-3/BCL2 immunostaining) evaluations, or subjected to biochemical assays to survey reactive oxygen species (ROS) and lipid peroxidation levels and total antioxidant capacity (TAC). The frequency of atretic follicles and number of cleaved-caspase-3-positive cells, as well as the ROS and lipid peroxidation levels, were significantly lower in ISO group compared to CTRL. A significantly higher number of BCL2-positive cells and TAC levels were also observed in ISO group. In conclusion, soy ISOs could decrease follicular atresia, apoptosis and oxidative stress, as well as increase the TAC in ovaries of female rats.
    Keywords:  apoptosis; ovarian aging; oxidative stress; rats; soy isoflavones
  6. Zhongguo Ying Yong Sheng Li Xue Za Zhi. 2018 Apr 08. 34(4): 289-293
    Sun WM, Zhang YZ, Wen X, Xi YX, Yuan D, Wang YH, Wei C, Xu CQ, Li HZ.
      OBJECTIVE: To investigate the recovery of protective effects of exogenous hydrogen sulfide (H2S) on hypoxia post-conditioning in aged H9C2 cells and its mechanism.METHODS: H9C2 cells (cardiomyocytes line) were treated with 30 μmol/L hydrogen peroxide (H2O2) for 2 hours, then cultured for 3 days in order to induce cellular aging. Aged H9C2 cells were randomly divided into 5 groups (n=8):Control group (Control), hypoxia/reoxygenation group (H/R), H/R + NaHS group, hypoxia post-conditioning (PC) group, PC+NaHS group. H/R model:the cells were exposed to hypoxic culture medium (serum and sugar free medium, pH=6.8) for 3 hours and then cultured at normal condition for 6 hours. PC model:at the end of hypoxia for 3 hours, the cells were exposed to normoxic culture solution for 5 minutes, then the cells were placed in hypoxic solution for 5 minutes, the cycle above-mentioned was repeated 3 times and followed by reoxygenation for 6 hours. Advanced glycation end products (AGEs) content and caspase-3 activity were detected by ELISA. The cell viability was observed by cell counting kit-8 (CCK-8). The reactive oxygen species (ROS) levels were analyzed using 2, 7-dichlorodihydrofluorescein diacetate (DCFH-DA) staining. The apoptotic rate was determined through Hoechst 33342 staining. The mRNA levels of relative gene expression were detected by real-time PCR.
    RESULTS: Thirty μmol/L H2O2 induced H9C2 cell senescence while did not lead to apoptosis. Compared with control group, cell viability was decreased, the apoptotic rate、levels of ROS and the mRNA of caspase-3, caspase-9 and Bcl-2 were increased in H/R and PC groups (P<0.01). There were no differences in the above indexes between PC group and H/R group. Supplementation of NaHS increased cell viability and decreased apoptotic rate and oxidative stress. The effects of PC + NaHS on the above indexes were better than those of H/R+NaHS group.
    CONCLUSIONS: Exogenous H2S can restore the protective effect of PC on the aged H9C2 cells, and its mechanism is related to the inhibition of oxidative stress and apoptosis.
    Keywords:  H9C2 cells; apoptosis; hydrogen sulfide; hypoxia post-conditioning; hypoxia/reoxygenation
  7. Neurotox Res. 2019 Feb 22.
    Ertilav K, Nazıroğlu M, Ataizi ZS, Braidy N.
      The rate of mitosis of cancer cells is significantly higher than normal primary cells with increased metabolic needs, which in turn enhances the generation of reactive oxygen species (ROS) production. Higher ROS production is known to increase cancer cell dependence on ROS scavenging systems to counteract the increased ROS. Therapeutic options which selectively modulate the levels of intracellular ROS in cancers are likely candidates for drug discovery. Docetaxel (DTX) has demonstrated antitumor activity in preclinical and clinical studies. It is thought that DTX induces cell death through excessive ROS production and increased Ca2+ entry. The Ca2+ permeable TRPM2 channel is activated by ROS. Selenium (Se) has been previously used to stimulate apoptosis for the treatment of glioblastoma cells resistant to DTX. However, the potential mechanism(s) of the additive effect of DTX on TRPM2 channels in cancer cells remains unclear. The aim of this study was to evaluate the effect of combination therapy of DTX and Se on activation of TRPM2 in DBTRG glioblastoma cells. DBTRG cells were divided into four treatment groups: control, DTX (10 nM for 10 h), Se (1 μM for 10 h), and DTX+Se. Our study showed that apoptosis (Annexin V and propidium iodide), mitochondrial membrane depolarization (JC1), and ROS production levels were increased in DBTRG cells following treatment with Se and DTX respectively. Cell number and viability, and the levels of apoptosis, JC1, ROS, and [Ca2+]i, induced by DTX, were further increased following addition of Se. We also observed an additive increase in the activation of the NAD-dependent DNA repair enzyme poly (ADP-ribose) polymerase-1 (PARP-1) activity, which was accompanied by a decline in its essential substrate NAD+. As well, the Se- and DTX-induced increases in intracellular Ca2+ florescence intensity were decreased following treatment with the TRPM2 antagonist N-(p-amylcinnamoyl) anthranilic acid (ACA). Therefore, combination therapy with Se and DTX may represent an effective strategy for the treatment of glioblastoma cells and may be associated with TRPM2-mediated increases in oxidative stress and [Ca2+]i.
    Keywords:  Apoptosis; Docetaxel; Glioblastoma; Selenium; TRPM2 channel
  8. Mol Cell Biochem. 2019 Feb 18.
    Ahmed HH, Taha FM, Omar HS, Elwi HM, Abdelnasser M.
      DN is recognized as not only a leading cause of end stage renal disease (ESRD) but also an independent risk factor for cardiovascular disease (CVD). Novel therapeutic approaches to diabetic nephropathy (DN) are needed, or else, healthcare resources will be overwhelmed by the expected worldwide increase in associated cases of ESRD and CVD. Reactive oxygen species (ROS) and advanced glycation end product (AGE) are implicated in the development of DN. Hydrogen sulfide (H2S) is known for its antioxidant and antiapoptotic characteristics. Simultaneously diabetics have lower H2S levels. Thus, it is worth investigating the use of H2S in treatment of DN. To investigate the potential therapeutic role of H2S in DN. Sixty male rats were divided into four groups: control, DN, DN+NaHS30 µmol/kg/day and DN+NaHS100 µmol/kg/day. Fasting blood sugar (FBS), kidney function tests, SIRT1 activity, superoxide dismutase activity (SOD), malondialdehyde (MDA) and expression of caspase3 and p53 in renal tissues were assessed. Kidney was examined histopathologically. DN rats had higher FBS, renal dysfunction, decreased SIRT1 and SOD activity levels, increased caspase3 and p53 relative expression and increased MDA in renal tissues. NaHS increased SIRT1 and reversed biochemical, apoptotic, oxidant and pathologic parameters characteristic of DN, with better results using a dose of 100 µmol/kg/day. H2S has a protective role against DN through decreasing FBS, ROS, apoptosis and upregulating SIRT1, thus preserving renal cells from further damage caused by DM.
    Keywords:  Caspase 3; Diabetic nephropathy; H2S; SIRT1; SOD
  9. Phytomedicine. 2018 Dec 17. pii: S0944-7113(18)30617-2. [Epub ahead of print]57 215-222
    Parzonko A, Kiss AK.
      BACKGROUND: Among solar radiation, ultraviolet light is the most harmful for the skin, because of intracellular reactive oxygen species formation, leading to oxidative stress, cell damage and apoptosis. Crucial role in skin protection against oxidative stress play antioxidant enzymes regulated by Nrf2 transcription factor. Some plant-derived polyphenols are known to protect skin fibroblasts against UV through induction of Nrf2-dependent antioxidant genes expression.PURPOSE: We previously found out that water extracts from Galinsoga sp. herb protected human dermal fibroblasts against UVA-induced oxidative stress and apoptosis. However, which compounds were responsible for such protective action remained unclear. Here, we investigated photoprotective potential and mechanism of action of two main isolated compounds, 2,3,5(2,4,5)-tricaffeoylaltraric acid and 2,4(3,5)-dicaffeoylglucaric acid, on human dermal fibroblasts (NHDF).
    STUDY DESIGN/METHODS: NHDF cells were pretreated with tested compounds (6.25-50 µM) and irradiated with UVA (25 J/cm2). Intracellular ROS and GSH level, cell viability, cell membrane integrity and apoptosis were measured. HO-1 protein expression and Nrf2 transcription factor activation were also assessed.
    RESULTS: Cells pretreated with tested compounds prior to UVA showed inhibition of intracellular ROS formation and increase of GSH level. Significant increase of cell viability was also observed, as well as decrease of LDH release and a the rate of apoptotic cells in comparison to untreated cells. Furthermore, tested compounds increased HO-1 expression and activated the Nrf2 transcription factor in NHDF cells.
    CONCLUSION: Present study demonstrated that caffeic acid derivatives present in Galinsoga parviflora herb, in particular tricaffeoylaltraric acid may protect dermal fibroblasts against UVA-induced oxidative stress through activation of intracellular antioxidative system. Such caffeic acid derivatives are bioactive compounds which might prevent UV-induced photoageing and photocarcinogenesis.
    Keywords:  Caffeic acid derivatives; Galinsoga parviflora; Nrf2; Oxidative stress; Skin fibroblasts; UVA
  10. Gene. 2019 Feb 14. pii: S0378-1119(19)30126-X. [Epub ahead of print]
    Dong RF, Tai LW, Zhang B, Shi FK, Liu HM, Duan PC, Cheng Y.
      Neuron damage contributes to ischemic brain injury. Although FMS-like tyrosine kinase-3 (FLT3) plays a critical role in neuron survival, its function and molecular mechanism in cerebral ischemia/reperfusion injury is unclear. In the present study, we exposed SH-SY5Y cells to oxygen and glucose deprivation/reoxygenation (OGD/R) to mimic ischemia/reperfusion injury. We found that FLT3 and MAPK14/p38α expression increased in OGD/R-treated cells. FLT3 silence significantly increased OGD/R-induced SH-SY5Y cell survival, inhibited reactive oxygen species production. Also, we observed that FLT3 silence suppressed OGD/R-induced SH-SY5Y cell apoptosis, apoptosis related protein Bax level and caspase-3 activity was decreased and Bcl-2 expression was increased in FLT3 silence SH-SY5Y cell treated with OGD/R. Furthermore, FLT3 depletion induced MAPK14/p38α inhibition in SH-SY5Y cultures after OGD/R exposure. These findings suggest that MAPK14/p38α overexpression reverses the action of FLT3 silence in OGD/R-induced SH-SY5Y cells. They also provide the first evidence that FLT3 silence has a neuroprotective role in OGD/R-induced SH-SY5Y cell damage. These data provide insight about potential neuroprotective molecular for ischemia/reperfusion injury.
    Keywords:  FMS-like tyrosine kinase-3; Ischemia/reperfusion injury; MAPK14/p38a; OGD/R
  11. Cancer Manag Res. 2019 ;11 1023-1032
    Zhang J, Geng H, Liu L, Zhang H.
      Background/Aims: Cytotoxicity induced by reactive oxygen species (ROS) is critical for the effectiveness of chemotherapeutic drugs used in the treatment of acute myeloid leukemia (AML). This study aimed to investigate whether ROS contributes to cytotoxicity in AML cells when treated with homoharringtonine (HHT) and etoposide (ETP) in combination.Methods: AML cell lines THP1 and HL60 and primary AML cells from patients were treated with HHT and ETP alone or in combination, and cell viability was determined by trypan blue exclusion test, and apoptosis was analyzed by annexin-V/propidium iodide double staining as well as Western blot for measuring expression of cleaved caspase-9 and cleaved caspase-3. Intracellular ROS level was detected by DCFH-DA fluorescence assay, and N-Acetyl-L-cysteine (NAC) was used to scavenge intracellular ROS. Retroviral infection was applied to mediate stable overexpression in AML cells.
    Results: We show that HHT and ETP exhibit synergistic cytotoxicity in AML cell lines and primary AML cells in vitro, and meanwhile, HHT causes elevated ROS generation in ETP-treated AML cells. We next reveal that the elevated ROS is a critical factor for the synergistic cytotoxicity, since ROS scavenge by NAC remarkably diminishes this effect. Mechanistically, we demonstrate that HHT causes elevated ROS generation by disabling thioredoxin-mediated antioxidant defense. Finally, similar to HHT treatment, depletion of thioredoxin sensitizes AML to ETP treatment.
    Conclusion: These results provide the foundation for augmenting the efficacy of ETP in treating AML with HHT, and also highlight the importance of targeting ROS in improving treatment outcome in AML.
    Keywords:  acute myeloid leukemia; antioxidant defense; etoposide; homoharringtonine; reactive oxygen species; synergistic cytotoxicity; thioredoxin
  12. Int J Mol Sci. 2019 Feb 13. pii: E804. [Epub ahead of print]20(4):
    Luparello C, Asaro DML, Cruciata I, Hassell-Hart S, Sansook S, Spencer J, Caradonna F.
      We examined the effects of the ferrocene-based histone deacetylase-3 inhibitor Pojamide (N¹-(2-aminophenyl)-N⁸-ferrocenyloctanediamide) and its two derivatives N¹-(2-aminophenyl)-N⁶-ferrocenyladipamide and N¹-(2-aminophenyl)-N⁸-ferroceniumoctanediamide tetrafluoroborate on triple-negative MDA-MB-231 breast cancer cells. Viability/growth assays indicated that only the first two compounds at 70 μM concentration caused an approximate halving of cell number after 24 h of exposure, whereas the tetrafluoroborate derivative exerted no effect on cell survival nor proliferation. Flow cytometric and protein blot analyses were performed on cells exposed to both Pojamide and the ferrocenyladipamide derivative to evaluate cell cycle distribution, apoptosis/autophagy modulation, and mitochondrial metabolic state in order to assess the cellular basis of the cytotoxic effect. The data obtained show that the cytotoxic effect of the two deacetylase inhibitors may be ascribed to the onset of non-apoptotic cell death conceivably linked to a down-regulation of autophagic processes and an impairment of mitochondrial function with an increase in intracellular reactive oxygen species. Our work expands the list of autophagy-regulating drugs and also provides a further example of the role played by the inhibition of autophagy in breast cancer cell death. Moreover, the compounds studied may represent attractive and promising targets for subsequent molecular modeling for anti-neoplastic agents in malignant breast cancer.
    Keywords:  apoptosis; autophagy; breast cancer cells; cell cycle; cell viability; histone deacetylase inhibitor; mitochondrial transmembrane potential; reactive oxygen species
  13. Free Radic Res. 2019 Feb 18. 1-221
    Ma J, Jin G.
      Alleviating the oxidant stress associated with myocardial ischemia reperfusion has been demonstrated as a potential therapeutic approach to limit ischemia reperfusion (I/R)-induced cardiac damage. It's reported that EGFR/erbB2 signaling is an important cardiac survival pathway in cardiac function and activation of EGFR has a cardiovascular effect in global ischemia. Epidermal growth factor (EGF), a typical EGFR ligand, was considered to have a significant role in activating EGFR. However, no evidence has been published whether exogenous EGF has protective effects on myocardial ischemia reperfusion. This study aims to investigate the effects of EGF in I/R-induced heart injury and to demonstrate its mechanisms. H9c2 cells challenged with H2O2 were used for in vitro biological activity and mechanistic studies. The malondialdehyde (MDA) and Superoxide Dismutase (SOD) levels in H9c2 cells were determined, and the cell viability was assessed by MTT assay. Myocardial I/R mouse administrated with or without EGF were used for in vivo studies. Pretreatment of H9c2 cells with EGF activated Nrf2 signaling pathway, attenuated H2O2-increased MDA and H2O2-reduced SOD level, followed by the inhibition of H2O2-induced cell death. In in vivo animal models of myocardial I/R, administration of EGF reduced infarct size and myocardial apoptosis. These data support that EGF decreases oxidative stress and attenuates myocardial ischemia reperfusion injury via activating Nrf2.
    Keywords:  Epidermal growth factor (EGF); Nrf2; myocardial ischemia reperfusion; reactive oxygen species (ROS)
  14. Int Immunopharmacol. 2019 Feb 19. pii: S1567-5769(19)30126-2. [Epub ahead of print]70 37-46
    Khosravi Z, Sedaghat R, Baluchnejadmojarad T, Roghani M.
      Diabetes mellitus (DM) is a prevalent metabolic disorder that is associated with development of some complications in male reproductive system including testicular damage, sexual dysfunction, abnormal spermatogenesis, and infertility. Diosgenin is a natural steroidal saponin with anti-diabetic, anti-oxidative, and anti-inflammatory effects. This research study was undertaken to explore the protective effect of diosgenin against diabetes-induced testicular damage in the rat. Ten days following streptozotocin (STZ; i.p.), diosgenin was daily administered for 6 weeks (p.o.). Diosgenin administration to diabetic rats significantly improved body weight and lowered serum glucose. In addition, diosgenin-treated diabetic group had a significantly lower level of malondialdehyde (MDA), protein carbonyl, greater level of glutathione (GSH), and higher activity of superoxide dismutase (SOD), catalase, and glutathione peroxidase (GPx) in addition to testicular improvement of ferric reducing antioxidant power (FRAP). Furthermore, diosgenin significantly improved serum insulin and testosterone level and alleviated testicular markers of inflammation including tumor necrosis factor α (TNFα) and interleukin 6 (IL-6) in diabetic rats. Moreover, apoptotic markers including caspase 3 activity, Annexin V, and DNA fragmentation decreased, mitochondrial membrane potential (MMP) accentuated, and myeloperoxidase (MPO) activity as a biomarker of neutrophil infiltration decreased in diosgenin-treated diabetic group. Additionally, diosgenin was capable to improve sperm count, motility, and viability in addition to prevention of damage to seminiferous tubules in diabetic animals. Collectively, diosgenin ameliorates testicular damage in DM, at least via partial suppression of apoptosis, oxidative stress, inflammation, and neutrophil infiltration and also via partial restoration of mitochondrial integrity.
    Keywords:  Apoptosis; Diabetes mellitus; Diosgenin; Inflammation; Oxidative stress; Testis
  15. Exp Ther Med. 2019 Mar;17(3): 1523-1528
    Zhang Q, Yang D.
      Emerging evidence has demonstrated the antitumor activity of allicin in various tumors. However, little study has been carried out on the functional role of allicin in cervical cancer. Our data showed that allicin suppressed cervical cancer cell viability in a time- and dose-dependent manner. Allicin treatment could reverse H2O2-induced reactive oxygen species accumulation. Meanwhile, levels of glutathione and superoxide dismutase were increased, but malondialdehyde was decreased after allicin incubation for 48 h. Furthermore, TUNEL staining showed that H2O2 treatment induced cell apoptosis, but allicin treatment could decrease cell apoptosis. Western blot assay showed that allicin could suppress the expression of nuclear factor erythroid 2-related factor 2 (NRF2) and heme oxygenase 1. We also showed that NRF2 prompted SiHa cell proliferation and reduced SiHa cell apoptosis. More importantly, allicin-inactivated phosphoinositide 3-kinase/protein kinase B (PI3K/AKT) signaling could be partially reversed by overexpressing of NRF2. We also evaluated cell apoptosis in SiHa cells transfected with plasmid NRF2. Our data showed that allicin-induced cell apoptosis (43.5±3.8%) could largely be abolished by upregulation of NRF2 (12.3±2.08%). In summary, our data showed allicin was effective in suppressing the malignant phenotype of cervical cancer cells mainly by inhibiting the expression of NRF2, showing the potential clinical benefits of allicin in cervical cancer patients.
    Keywords:  allicin; cervical cancer; nuclear factor erythroid 2-related factor 2; oxidative stress
  16. Exp Ther Med. 2019 Mar;17(3): 1670-1676
    Qi Y, Ding Z, Yao Y, Ma D, Ren F, Yang H, Chen A.
      Ovarian cancer is one of the main causes of cancer-associated mortality across the world. Currently, ovarian cancer is mainly treated with chemotherapy. However, ovarian cancer is detected at advanced stages and chemotherapy has numerous side effects. In addition, the results of current chemotherapy on the treatment of ovarian cancer are less than satisfactory. Therefore, there is an urgent need to develop novel and more viable chemotherapeutic agents that can be used to treat ovarian cancer. The present study was designed to synthesize a series of novel triazole analogs of the bioactive apigenin-7-methyl ether to evaluate its anticancer activity against three human ovarian cancer cell lines. A total of eight novel triazole derivatives were synthesized and screened for their anticancer activity. Of all the derivatives, a derivative named 3d exhibited significant and dose-dependent anticancer activity against the SKOV3 ovarian cancer cell line. The IC50 of 3d was found to be 10 µM against the SKOV3 cancer cell line. It was also observed that 3d induced apoptosis in SKOV3 cancer cells through the accretion of reactive oxygen species and reduction in mitochondrial membrane potential. The molecule also modulated the expression of B-cell lymphoma 2 (Bcl-2) and Bcl-2-associated X protein. Taken together, these results showed that the apigenein-7-methyl ether novel derivative 3d may prove an important lead molecule for the treatment of ovarian cancer.
    Keywords:  apigenein-7-methyl ether; apoptosis; ovarian cancer; reactive oxygen species
  17. Int J Nanomedicine. 2019 ;14 921-936
    Song L, Zhang W, Chen H, Zhang X, Wu H, Ma M, Wang Z, Gu N, Zhang Y.
      Background: Cancer targeting nanoprobes with precisely designed physicochemical properties may show enhanced pharmacological targeting and therapeutic efficacy. As a widely used commercialized antibody, rituximab has been in clinical use for three decades and has lengthened or even saved thousands of lives. However, many people cannot benefit from rituximab treatment because of drug resistance or side effects.Methods: In this study, a 13-nm rituximab-conjugated magnetic nanoparticle was developed as a therapeutic nanoprobe targeting CD20 overexpressing malignant lymphoma cells to enhance the treatment effects of rituximab. The magnetic cores (2,3-dimercaptosuccinicacid modified Fe3O4 nanoparticles, Fe3O4@DMSA) of the nanoprobes with an average diameter of 6.5 nm were synthesized using a co-precipitation method. Rituximab was then conjugated on the surface of Fe3O4@DMSA using a cross-linking agent (carbodiimide/N-hydroxysulfosuccinimide sodium salt). Based on theoretical calculations, approximately one antibody was coupled with one nanoparticle, excluding the multivalent antibody effect.
    Results: Cell targeting experiments and magnetic resonance (MR) signal and T2 measurements showed that the Fe3O4@DMSA@Ab nanoprobes have specific binding affinity for CD20-positive cells. Compared to rituximab and Fe3O4@DMSA, Fe3O4@DMSA@Ab nanoprobes significantly reduced cell viability and promoted Raji cell apoptosis. Initiating events of apoptosis, including increased intracellular calcium and reactive oxygen species, were observed in nanoprobe-treated Raji cells. Nanoprobe-treated Raji cells also showed the most drastic decrease in mitochondrial membrane potential and Bcl-2 expression, compared to rituximab and Fe3O4@DMSA-treated Raji cells.
    Conclusion: These results indicate that Fe3O4@DMSA@Ab nanoprobes have the potential to serve as MRI tracers and therapeutic agents for CD20-positive cells.
    Keywords:  CD20-targeted nanoprobes; MMP; ROS; calcium; cell apoptosis; mitochondrial membrane potential; reactive oxygen species
  18. J Cell Physiol. 2019 Feb 19.
    Lin Y, Li B, Zhao J, Wei L, Wang Y, Wang M, Dia VP, Meng X.
      The use of natural compounds to potentiate the effect of drugs and lower their adverse effects is an active area of research. The objective is to determine the effect of combined blueberry extracts (BE) and oxaliplatin (OX) in colon cancer cells. The results demonstrated that treatments of BE/OX showed inhibitory effects on HCT-116 cell and nontoxic effect on CCD-18Co normal colon cells. Flow cytometry analysis indicated that treatment with the BE, OX or in combination could induce G0/G1 cell cycle arrest, apoptosis, increase of reactive oxygen species, and induce loss of mitochondrial membrane potential in HCT-116 cells. Furthermore, after treatments, the expression of inflammatory cytokines was decreased, cyclin D1 and CDK4 were decreased; caspases-3 and 9 were activated; the Akt/Bad/Bcl-2 pathway was modulated. Moreover, the combination treatment had a considerably higher growth inhibitory effect on human colon cancer HCT-116 cells than that of BE or oxaliplatin alone. Our results showed that BE increased the anticolon cancer effect of OX making it an attractive strategy as adjuvant therapy to potentially reduce the adverse side effects associated with chemotherapeutic drugs.
    Keywords:  apoptosis; blueberry extracts; cell cycle; colon cancer; oxaliplatin
  19. J Fungi (Basel). 2019 Feb 13. pii: E16. [Epub ahead of print]5(1):
    Govindharaj M, Arumugam S, Nirmala G, Bharadwaj M, Murugiyan K.
      The aim of the present study is to extract the bioactive compounds which can induce the apoptosis in breast cancer cell line MCF-7 by marine basidiomycetes. Internal Transcribed Spacer (ITS) sequences based molecular taxonomic study confirmed that collected the marine basidiomycetes belongs to Fulvifomes sp. Further, the isolated compounds from the Fulvifomes sp. confirmed as ergosterol peroxide (EP) by spectroscopic studies. The compound inhibited 50% of the cell growth (IC50) at the concentration of 40 µg/mL and induced 90% cell death (IC 90) at the concentration of 80 µg/mL. The ergosterol peroxide generated Reactive Oxygen Species (ROS) and induced apoptotic cell death in MCF-7. Ethidium bromide/Acridine Orange (Et/Br) staining showed the increased number of early and late apoptosis in treated MCF-7 cells. The compounds treated cells indicated the significant loss of mitochondrial membrane potential (Δψm) with p < 0.05. The induction of apoptosis by marine basidiomycetes derived ergosterol peroxide was confirmed by chromatin condensation in MCF7 cells using Hoechst staining 33342.
    Keywords:  Marine Basidiomycetes; apoptosis induction; breast cancer; ergosterol peroxide
  20. Int J Prev Med. 2019 ;10 5
    Hamzeh M, Hosseinimehr SJ, Karimpour A, Mohammadi HR, Khalatbary AR, Talebpour Amiri F.
      Background: Cyclophosphamide (CP), as a chemotherapy drug, causes severe damage in testicular tissue through producing free radicals. Cerium oxide nanoparticles (NC) exhibit antioxidant and anti-inflammatory properties. The purpose of this study was to investigate the protective effect of NC on CP-induced testicular damage in mice.Methods: In this experimental study, thirty-two male mice were divided into four groups (eight mice in each group). The control group was received intraperitoneally (IP) normal saline, NC group was received NC for three consecutive days (100 μg/kg, IP), CP group was received CP (200 mg/kg, IP), and the CP + NC group received NC, three consecutive days before receiving CP. After 2 days, testicles were assessed for biochemical, histomorphometrical, histopathological, and immunohistochemical analyses.
    Results: CP administration caused statistically significant increases in sperm abnormality, malondialdehyde, protein carbonyl levels, reactive oxygen species, level and apoptosis, and decreases in sperm count, sperm viability, testosterone, glutathione activity, the mean thickness of the germinal epithelium, diameter of seminiferous tubules in mice. Degeneration, necrosis, arrest of spermatogenesis, congestion, and atrophy in testicular tissue confirmed the low Johnsen's Testicular score in CP group. Administration of NC significantly ameliorated the CP-induced adverse effects on testis compared with the CP group. In addition, pretreatment mice with NC significantly reduced caspase-3 immunoreactivity induced by CP in testis.
    Conclusions: This study showed that NC with scavenging free radicals and antiapoptotic properties enable to reduce the side effects of CP in the testicular tissue.
    Keywords:  Caspase-3; cerium oxide; cyclophosphamide; oxidative stress; testis; toxicity
  21. J Mol Cell Cardiol. 2019 Feb 13. pii: S0022-2828(18)31153-2. [Epub ahead of print]
    Meijles DN, Zoumpoulidou G, Markou T, Rostron KA, Patel R, Lay K, Handa BS, Wong B, Sugden PH, Clerk A.
      Reactive oxygen species (ROS) play a key role in development of heart failure but, at a cellular level, their effects range from cytoprotection to induction of cell death. Understanding how this is regulated is crucial to develop novel strategies to ameliorate only the detrimental effects. Here, we revisited the fundamental hypothesis that the level of ROS per se is a key factor in the cellular response by applying different concentrations of H2O2 to cardiomyocytes. High concentrations rapidly reduced intracellular ATP and inhibited protein synthesis. This was associated with activation of AMPK which phosphorylated and inhibited Raptor, a crucial component of mTOR complex-1 that regulates protein synthesis. Inhibition of protein synthesis by high concentrations of H2O2 prevents synthesis of immediate early gene products required for downstream gene expression, and such mRNAs (many encoding proteins required to deal with oxidant stress) were only induced by lower concentrations. Lower concentrations of H2O2 promoted mTOR phosphorylation, associated with differential recruitment of some mRNAs to the polysomes for translation. Some of the upregulated genes induced by low H2O2 levels are cytoprotective. We identified p21Cip1/WAF1 as one such protein, and preventing its upregulation enhanced the rate of cardiomyocyte apoptosis. The data support the concept of a "redox rheostat" in which different degrees of ROS influence cell energetics and intracellular signalling pathways to regulate mRNA and protein expression. This sliding scale determines cell fate, modulating survival vs death.
    Keywords:  Cytoprotection; Immediate early genes; Oxidative stress; Protein synthesis; Raptor; mTOR; p21(Cip1/WAF1)
  22. Exp Biol Med (Maywood). 2019 Feb 20. 1535370219831225
    Mercantepe F, Topcu A, Rakici S, Tumkaya L, Yilmaz A.
      IMPACT STATEMENT: Some six million cancer patients currently receive radiotherapy. Radiotherapy eliminates cancer cells by accelerating their death. However, radiotherapy is not selective, and it therefore harms healthy tissues around cancerous tissue. The latest studies have shown that the irradiation of biological materials causes a rapid increase in reactive oxygen species (ROS) in the tissue as a result of exposure of the target molecule to direct and indirect ionization. N-acetylcysteine (NAC) is an antioxidant that permits the elimination of free oxygen radicals and that contributes to glutathione synthesis. Our study, therefore, examined the effects of radiation resulting from radiotherapy on the small intestine at the molecular level, and prospectively considered the potential protective characteristics of NAC against gastrointestinal syndrome resulting from radiotherapy.
    Keywords:  Apoptosis; N-acetylcysteine; caspase-3; ionizing radiation; oxidative stress; rat
  23. J Am Heart Assoc. 2019 Feb 19. 8(4): e011134
    Hijmans JG, Stockelman KA, Garcia V, Levy MV, Brewster LM, Bammert TD, Greiner JJ, Stauffer BL, Connick E, DeSouza CA.
      Background Circulating microparticles have emerged as biomarkers and effectors of vascular disease. Elevated rates of cardiovascular disease are seen in HIV -1-seropositive individuals. The aims of this study were to determine: (1) if circulating microparticles are elevated in antiretroviral therapy-treated HIV -1-seropositive adults; and (2) the effects of microparticles isolated from antiretroviral therapy -treated HIV -1-seropositive adults on endothelial cell function, in vitro. Methods and Results Circulating levels of endothelial-, platelet-, monocyte-, and leukocyte-derived microparticles were determined by flow cytometry in plasma from 15 healthy and 15 antiretroviral therapy-treated, virologically suppressed HIV -1-seropositive men. Human umbilical vein endothelial cells were treated with microparticles from individual subjects for 24 hours; thereafter, endothelial cell inflammation, oxidative stress, senescence, and apoptosis were assessed. Circulating concentrations of endothelial-, platelet-, monocyte-, and leukocyte-derived microparticles were significantly higher (≈35%-225%) in the HIV -1-seropositive compared with healthy men. Microparticles from HIV -1-seropositive men induced significantly greater endothelial cell release of interleukin-6 and interleukin-8 (≈20% and ≈35%, respectively) and nuclear factor-κB expression while suppressing anti-inflammatory microRNAs (miR-146a and miR-181b). Intracellular reactive oxygen species production and expression of reactive oxygen species -related heat shock protein 70 were both higher in cells treated with microparticles from the HIV -1-seropositive men. In addition, the percentage of senescent cells was significantly higher and sirtuin 1 expression lower in cells treated with HIV -1-related microparticles. Finally, caspase-3 was significantly elevated by microparticles from HIV -1-seropositive men. Conclusions Circulating concentrations of endothelial-, platelet-, monocyte-, and leukocyte-derived microparticles were higher in antiretroviral therapy-treated HIV -1-seropositive men and adversely affect endothelial cells promoting cellular inflammation, oxidative stress, senescence, and apoptosis. Circulating microparticles may contribute to the vascular risk associated with HIV -1 infection.
    Keywords:  HIV‐1; endothelial dysfunction; inflammation; microRNA; microparticles
  24. Cell Physiol Biochem. 2019 ;52(1): 119-140
    Smolková B, Lunova M, Lynnyk A, Uzhytchak M, Churpita O, Jirsa M, Kubinová Š, Lunov O, Dejneka A.
      BACKGROUND/AIMS: Alteration of cancer cell redox status has been recognized as a promising therapeutic implication. In recent years, the emerged field of non-thermal plasma (NTP) has shown considerable promise in various biomedical applications, including cancer therapy. However, understanding the molecular mechanisms procuring cellular responses remains incomplete. Thus, the aim of this study was a rigorous biochemical analysis of interactions between NTP and liver cancer cells.METHODS: The concept was validated using three different cell lines. We provide several distinct lines of evidence to support our findings; we use various methods (epifluorescent and confocal microscopy, clonogenic and cytotoxicity assays, Western blotting, pharmacological inhibition studies, etc.).
    RESULTS: We assessed the influence of NTP on three human liver cancer cell lines (Huh7, Alexander and HepG2). NTP treatment resulted in higher anti-proliferative effect against Alexander and Huh7 relative to HepG2. Our data clearly showed that the NTP-mediated alternation of mitochondrial membrane potential and dynamics led to ROS-mediated apoptosis in Huh7 and Alexander cells. Interestingly, plasma treatment resulted in p53 down-regulation in Huh7 cells. High levels of Bcl-2 protein expression in HepG2 resulted in their resistance in response to oxidative stress- mediated by plasma.
    CONCLUSION: We show thoroughly time- and dose-dependent kinetics of ROS accumulation in HCC cells. Furthermore, we show nuclear compartmentalization of the superoxide anion triggered by NTP. NTP induced apoptotic death in Huh7 liver cancer cells via simultaneous downregulation of mutated p53, pSTAT1 and STAT1. Contrary, hydrogen peroxide treatment results in autophagic cell death. We disclosed detailed mechanisms of NTP-mediated alteration of redox signalling in liver cancer cells.
    Keywords:  Apoptosis, Non-thermal plasma; Oxidative stress; Reactive oxygen species; Stress resistance; p53
  25. Ecotoxicol Environ Saf. 2019 Feb 13. pii: S0147-6513(19)30121-6. [Epub ahead of print]173 131-141
    Yu H, Zhang J, Ji Q, Yu K, Wang P, Song M, Cao Z, Zhang X, Li Y.
      The present study aimed to investigate whether melatonin (MT) treatment can attenuate immunotoxicity induced by aluminum chloride (AlCl3) in rat spleen. Forty-eight healthy male Wistar rats were randomly allocated and treated with AlCl3 and/or MT. Rats were orally administered with AlCl3 for 90 days, from 61st days, rats were injected intraperitoneally with MT for 30 days. Firstly, we found that MT relieved the AlCl3-induced immunosuppression by improving spleen structural damage, CD3+ and CD4+ T lymphocyte subsets, IL-2 and TNF-α mRNA expressions and decreasing CD8+ T lymphocyte subsets. Secondly, MT attenuated the AlCl3-induced oxidative stress in rat spleen by decreasing the levels of ROS and MDA, while increasing the activities of SOD and CAT. Thirdly, MT relieved the AlCl3-induced apoptosis in rat spleen by increasing the MMP and Bcl-2 mRNA and protein expressions, while decreasing apoptosis rates, activity of Caspase-3 and pro-apoptotic gene expression. Finally, MT increased Nrf2 nuclear translocation, and Nrf2 target genes (HO-1, NQO1, SOD1 and CAT) mRNA expressions in the spleen of AlCl3-exposed rat. These results suggest that MT may alleviate AlCl3-induced immunotoxicity by inhibiting oxidative stress and apoptosis associated with the activation of Nrf2 signaling pathway, which could lay the foundation for the treatment of AlCl3 immunotoxicity.
    Keywords:  Aluminum chloride; Apoptosis; Immunotoxicity; Melatonin; Nrf2; Oxidative stress
  26. Mol Cancer Ther. 2019 Feb 20. pii: molcanther.0196.2018. [Epub ahead of print]
    Kim BR, Jeong YA, Jo MJ, Park SH, Na YJ, Kim JL, Jeong S, Yun HK, Kang SH, Lee DH, Oh SC.
      Despite an increase in the survival rate of cancer patients owing to the use of current chemotherapeutic agents, adverse effects of cancer therapies remain a concern. Combination therapies have been developed to increase efficacy, reduce adverse effects, and overcome drug resistance. Genipin is a natural product derived from Gardenia jasminoides, which has been associated with anti-inflammatory, anti-angiogenic, and anti-proliferative effects; hypertension; and anti-ischemic brain injuries. However, the enhancement of oxaliplatin sensitivity by genipin remains unexplored. Our study showed that a combination of genipin and oxaliplatin exerts synergistic anti-tumor effects in vitro and in vivo in colorectal cancer (CRC) cell lines through the reactive oxygen species (ROS)/endoplasmic reticulum (ER) stress/BIM pathway. Importantly, the combination did not affect normal colon cells. BIM knockdown markedly inhibited apoptosis induced by the combination. Additionally, genipin induced ROS by inhibiting superoxide dismutase 3 activity. These findings suggest that genipin may be a novel agent for increasing the sensitivity of oxaliplatin against CRC. The combination of oxaliplatin and genipin hold significant therapeutic potential with minimal adverse effects.
  27. Chemosphere. 2019 Feb 07. pii: S0045-6535(19)30238-3. [Epub ahead of print]223 263-274
    Wang R, Chen R, Wang Y, Chen L, Qiao J, Bai R, Ge G, Qin G, Chen C.
      Particulate matter (PM) exposure poses many adverse effects on human health. However, it is challenging to clearly differentiate between the contributions of individual pollutants on toxicity from complex mixtures of ambient air pollutants. The aim of this study is to generate aerosols constituted by silica nanoparticles (NPs) and bisulfate to serve as simulators of particle-associated high-sulfur air pollution. Then, the health impacts of sulfur dioxide were evaluated at the cellular level using an air-liquid interface (ALI) exposure chamber. BEAS-2B cells were exposed to either nano-silica or bisulfite aerosol individually or bisulfate-coated silica (SiO2-NH2@HSO3) for 3 h using the ALI. The cellular toxicities were carefully compared based on the exposure dosages. The ALI exposure of SiO2 NPs alone did not produce any apparent cytotoxicity in cells, but the aerosol exposure of SiO2-NH2@HSO3 significantly decreased the cell viability and enhanced the production of cellular reactive oxygen species in a dose-dependent manner. Consequently, the excessive oxidative stress resulted in mitochondrial damage as well as cellular apoptosis. ALI exposure can possibly reflect the realistic physiological exposure condition of the human respiratory system. As a derivative of the sulfur dioxide component of air pollution, sulfate exacerbates the toxic effects of inhalable PMs. This result may be due to the large surface area of the nanoparticles, with the possibility of carrying more sulfite to the target cells during aerosol exposure. The sulfate levels offer a meaningful complement to the present PM2.5 index of air pollution for achieving better human health protection.
    Keywords:  Aerosol components; Air-liquid interface; Nanoparticles; Silica; Toxicity
  28. Bull Exp Biol Med. 2019 Feb 20.
    Nakvasina MA, Popova LI, Lidokhova OV, Artyukhov VG.
      The effects of ROS on functional properties (cytotoxic activity, antibody-producing activity, TNFα synthesis, and free cytosol calcium level), membrane structure (by expression of some surface markers), and apoptosis of lymphocytic cells were estimated in the peripheral blood of healthy volunteers. 1O2, [Formula: see text], OH•, and H2O2 mostly suppressed cytotoxic activity of lymphocytes against Ehrlich ascites carcinoma cells and inhibited IgG synthesis and expression of receptors and surface markers (Fc receptors, CD3, CD19, and CD56). The exposure of lymphocytes to H2O2 (10-6 M), 1O2, and OH• was followed by an increase in the level of a secondary messenger, intracellular calcium, in comparison with non-exposed cells. The presence of exogenous calcium in the medium for lymphocyte suspending induced an increase in the number of cells at early and late stages of apoptosis 6 h after exposure to H2O2 and 1O2 in comparison with lymphocytes incubated in Ca2+-free medium.
    Keywords:  Ca2+; apoptosis; lymphocytes; reactive oxygen forms; structural and functional properties
  29. Nat Nanotechnol. 2019 Feb 18.
    Gong N, Ma X, Ye X, Zhou Q, Chen X, Tan X, Yao S, Huo S, Zhang T, Chen S, Teng X, Hu X, Yu J, Gan Y, Jiang H, Li J, Liang XJ.
      Mitochondrial redox homeostasis, the balance between reactive oxygen species and antioxidants such as glutathione, plays critical roles in many biological processes, including biosynthesis and apoptosis, and thus is a potential target for cancer treatment. Here, we report a mitochondrial oxidative stress amplifier, MitoCAT-g, which consists of carbon-dot-supported atomically dispersed gold (CAT-g) with further surface modifications of triphenylphosphine and cinnamaldehyde. We find that the MitoCAT-g particles specifically target mitochondria and deplete mitochondrial glutathione with atomic economy, thus amplifying the reactive oxygen species damage caused by cinnamaldehyde and finally leading to apoptosis in cancer cells. We show that imaging-guided interventional injection of these particles potently inhibits tumour growth in subcutaneous and orthotopic patient-derived xenograft hepatocellular carcinoma models without adverse effects. Our study demonstrates that MitoCAT-g amplifies the oxidative stress in mitochondria and suppresses tumour growth in vivo, representing a promising agent for anticancer applications.
  30. Drug Chem Toxicol. 2019 Feb 19. 1-7
    Wu D, Zhou WY, Lin XT, Fang L, Xie CM.
      OBJECTIVE: Bufalin has been reported to kill various types of cancer including human colorectal cancer. Our previous study demonstrated that bufalin induced cell death via autophagy in HT-29 and Caco-2 colon cancer cells, but the action of bufalin remains unclear. This study was conducted to investigate the role of bufalin in other colon cancer HCT-116 and SW620 cells as well as its potential mechanism.METHODS: The effect of bufalin in HCT-116 and SW620 colon cancer cells was detected by assessing cell viability and cell death. Apoptotic cells were analyzed by Western blot and trypan blue dye exclusion assay. Mitochondrial ROS production was analyzed by flow cytometry after DCFDA and DHR-123 staining. The potential mechanism was investigated via pharmacological inhibitors.
    RESULTS: Bufalin had high potency against HCT-116 and SW620 cells with IC50 values of 12.823 ± 1.792 nM and 26.303 ± 2.498 nM in HCT-116 and SW620 cells, respectively. Bufalin decreased cell viability, increased cell death as well as caspase-3 downstream target (cleaved PARP) accumulation, and these actions were significantly blocked by pan-caspase inhibitor zVAD-FMK. Mechanistically, ROS production, but neither the NAD(P)H oxidase, AMPK, ERK nor p38, is responsible for bufalin-induced apoptotic cell death. Moreover, bufalin-induced ROS generation is derived from mitochondria.
    CONCLUSION: Bufalin significantly induces apoptosis in HCT-116 and SW620 colon cancer cells via mitochondrial ROS-mediated caspase-3 activation. We believe that our novel findings will greatly alter our current understanding on the anti-cancer mechanism of bufalin in colon cancer cells and will pave the way for further exploiting the clinical application.
    Keywords:  Bufalin; ROS; apoptosis; colon cancer
  31. J Cell Physiol. 2019 Feb 19.
    Zhao X, Zeng Z, Gaur U, Fang J, Peng T, Li S, Zheng W.
      Metformin, a first line anti type 2 diabetes drug, has recently been shown to extend lifespan in various species, and therefore, became the first antiaging drug in clinical trial. Oxidative stress due to excess reactive oxygen species (ROS) is considered to be an important factor in aging and related disease, such as Alzheimer's disease (AD). However, the antioxidative effects of metformin and its underlying mechanisms in neuronal cells is not known. In the present study, we showed that metformin, in clinically relevant concentrations, protected neuronal PC12 cells from H2 O2 -induced cell death. Metformin significantly ameliorated cell death due to H2 O2 insult by restoring abnormal changes in nuclear morphology, intracellular ROS, lactate dehydrogenase, and mitochondrial membrane potential induced by H2 O2 . Hoechst staining assay and flow cytometry analysis revealed that metformin significantly reduced the apoptosis in PC12 cells exposed to H2 O2 . Western blot analysis further demonstrated that metformin stimulated the phosphorylation and activation of AMP-activated protein kinase (AMPK) in PC12 cells, while application of AMPK inhibitor compound C, or knockdown of the expression of AMPK by specific small interfering RNA or short hairpin RNA blocked the protective effect of metformin. Similar results were obtained in primary cultured hippocampal neurons. Taken together, these results indicated that metformin is able to protect neuronal cells from oxidative injury, at least in part, via the activation of AMPK. As metformin is comparatively cheaper with much less side effects in clinic, our findings support its potential to be a drug for prevention and treatment of aging and aging-related diseases.
    Keywords:  AMP-activated protein kinase; PC12 cells; aging; hydrogen peroxide; metformin
  32. J Cell Physiol. 2019 Feb 20.
    An Q, Peng W, Cheng Y, Lu Z, Zhou C, Zhang Y, Su J.
      Oocyte quality, which is directly related to reprogramming competence, is a major important limiting factor in animal cloning efficiency. Compared with oocytes matured in vivo, in vitro matured oocytes exhibit lower oocyte quality and reprogramming competence primarily because of their higher levels of reactive oxygen species. In this study, we investigate whether supplementing the oocyte maturation medium with melatonin, a free radical scavenger, could improve oocyte quality and reprogramming competence. We found that 10-9 M melatonin effectively alleviated oxidative stress, markedly decreased early apoptosis levels, recovered the integrity of mitochondria, ameliorated the spindle assembly and chromosome alignment in oocytes, and significantly promoted subsequent cloned embryo development in vitro. We also analyzed the effects of melatonin on epigenetic modifications in bovine oocytes. Melatonin increased the global H3K9 acetylation levels, reduced the H3K9 methylation levels, and minimally affected DNA methylation and hydroxymethylation. Genome-wide expression analysis of genes in melatonin-treated and nontreated oocytes was also conducted by high-throughput RNA sequencing. Our results indicated that melatonin ameliorates oocyte oxidative stress and improves subsequent in vitro development of bovine cloned embryos.
    Keywords:  cattle; melatonin; oocyte development; reactive oxygen species (ROS); somatic cell nuclear transfer (SCNT)
  33. Nat Commun. 2019 Feb 19. 10(1): 845
    Guo L, Cui C, Zhang K, Wang J, Wang Y, Lu Y, Chen K, Yuan J, Xiao G, Tang B, Sun Y, Wu C.
      Cell metabolism is strongly influenced by mechano-environment. We show here that a fraction of kindlin-2 localizes to mitochondria and interacts with pyrroline-5-carboxylate reductase 1 (PYCR1), a key enzyme for proline synthesis. Extracellular matrix (ECM) stiffening promotes kindlin-2 translocation into mitochondria and its interaction with PYCR1, resulting in elevation of PYCR1 level and consequent increase of proline synthesis and cell proliferation. Depletion of kindlin-2 reduces PYCR1 level, increases reactive oxygen species (ROS) production and apoptosis, and abolishes ECM stiffening-induced increase of proline synthesis and cell proliferation. In vivo, both kindlin-2 and PYCR1 levels are markedly increased in lung adenocarcinoma. Ablation of kindlin-2 in lung adenocarcinoma substantially reduces PYCR1 and proline levels, and diminishes fibrosis in vivo, resulting in marked inhibition of tumor growth and reduction of mortality rate. Our findings reveal a mechanoresponsive kindlin-2-PYCR1 complex that links mechano-environment to proline metabolism and signaling, and suggest a strategy to inhibit tumor growth.
  34. Nanoscale Res Lett. 2019 Feb 18. 14(1): 58
    Jang HH, Park SB, Hong JS, Lee HL, Song YH, Kim J, Jung YH, Kim C, Kim DM, Lee SE, Jeong YI, Kang DH.
      BACKGROUND: The aim of this study is to fabricate drug-eluting gastrointestinal (GI) stent using reactive oxygen species (ROS)-sensitive nanofiber mats for treatment of cholangiocarcinoma (CCA) cell. A ROS-producing agent, piperlongumine (PL)-incorporated nanofiber mats were investigated for drug-eluting stent (DES) application.METHODS: Selenocystamine-conjugated methoxy poly(ethylene glycol) (MePEG) was conjugated with poly(L-lactide) (PLA) to produce block copolymer (LEse block copolymer). Various ratios of poly(ε-caprolactone) (PCL) and LEse block copolymer were dissolved in organic solvent with PL, and then nanofiber mats were fabricated by electro-spinning techniques.
    RESULTS: The higher amount of LEse in the blend of PCL/LEse resulted in the formation of granules while PCL alone showed fine nanofiber structure. Nanofiber mats composed of PCL/LEse polymer blend showed ROS-sensitive drug release, i.e., PL release rate from nanofiber mats was accelerated in the presence of hydrogen peroxide (H2O2) while nanofiber mats of PCL alone have small changes in drug release rate, indicating that PL-incorporated nanofiber membranes have ROS responsiveness. PL itself and PL released from nanofiber mats showed almost similar anticancer activity against various CCA cells. Furthermore, PL released from nanofiber mats properly produced ROS generation and induced apoptosis of CCA cells as well as PL itself. In HuCC-T1 cell-bearing mice, PL-incorporated nanofiber mats showed improvement in anticancer activity.
    CONCLUSION: PL-incorporated ROS-sensitive nanofiber mats were coated onto GI stent and showed improved anticancer activity with ROS responsiveness. We suggested PL-incorporated ROS-sensitive nanofiber mats as a promising candidate for local treatment of CCA cells.
    Keywords:  Cholangiocarcinoma; Drug-eluting stent; GI stent; Nanofiber; Piperlongumine
  35. Bioorg Med Chem. 2019 Feb 04. pii: S0968-0896(18)31799-1. [Epub ahead of print]
    Sreenivasulu R, Reddy KT, Sujitha P, Kumar CG, Raju RR.
      In recent years, indole-indazolyl hydrazide-hydrazone derivatives with strong cell growth inhibition and apoptosis induction characteristics are being strongly screened for their cancer chemo-preventive potential. In the present study, N-methyl and N,N-dimethyl bis(indolyl)hydrazide-hydrazone analog derivatives were designed, synthesized and allowed to evaluate for their anti-proliferative and apoptosis induction potential against cervical (HeLa), breast (MCF-7 and MDA-MB-231) and lung (A549) cancer cell lines relative to normal HEK293 cells. The MTT assay in conjunction with mitochondrial potential assays and the trypan blue dye exclusion were employed to ascertain the effects of the derivatives on the cancer cells. Further, mechanistic studies were conducted on compound 14a to understand the biochemical mechanisms and functional interactions with various signaling pathways triggered in HeLa and MCF-7 cells. Compound 14a induced apoptosis via caspase independent pathway through the participation of mitogen-activated protein kinases (MAPK) such as extracellular signal related kinase (ERK) and p38 as well as p53 pathways. It originates the activation of pro-apoptotic proteins such as Bak and Mcl-1s and also strongly induced the generation of reactive oxygen species. In downstream signaling pathway, activated p53 protein interacted with MAPK pathways, including SAPK/c-Jun N-terminal protein kinase (JNK), p38 and ERK kinases resulting in apoptotic cell death. The involvement of MAPK cascades such as p38, ERK and p38 on compound 14a induced apoptotic cell death was evidenced by the fact that the inclusion of specific inhibitors of p38, ERK1/2 and JNK MAPK (SB2035809, PD98059 and SP600125) prevented the compound 14a towards induced apoptosis. The results clearly showed that MAP kinase cascades were crucial for apoptotic response in compound 14a induced cellular killing and were dependent on p53 activity. Based on the results, compound 14a was identified as a promising candidate for cancer therapeutics and these findings furnish a basis for further in vivo experiments on anti-proliferative activity.
    Keywords:  Anti-proliferative; Apoptosis; Bis(indole)hydrazide-hydrazone; MTT assay; Mechanistic studies
  36. Biomed Pharmacother. 2019 Feb 18. pii: S0753-3322(18)37096-3. [Epub ahead of print]112 108586
    Polloni L, Seni Silva AC, Teixeira SC, Azevedo FVPV, Zóia MAP, da Silva MS, Lima PMAP, Correia LIV, do Couto Almeida J, da Silva CV, Rodrigues Ávila VM, Goulart LRF, Morelli S, Guerra W, Oliveira Júnior RJ.
      This work reports the biological evaluation of a copper complex of the type [Cu(O-O)(N-N)ClO4], in which O-O = 4,4,4-trifluoro-1-phenyl-1,3-butanedione (Hbta) and N-N = 1,10-phenanthroline (phen), whose generic name is CBP-01. The cytotoxic effect of CBP-01 was evaluated by resazurin assay and cell proliferation was determined by MTT assay. DNA fragmentation was analyzed by gel electrophoresis. Cell cycle progression was detected through propidium iodide (PI) staining. Apoptosis and autophagy were determined by, respectively, Annexin V and 7-AAD staining and monodansylcadaverine (MDC) staining. The changes in intracellular reactive oxygen species levels were detected by DCFDA analysis. The copper complex CBP-01 showed in vitro antitumor activity with IC50s values of 7.4 μM against Sarcoma 180 and 26.4 against murine myoblast cells, displaying selectivity toward the tumor cell tested in vitro (SI > 3). An increase in reactive oxygen species (ROS) generation was observed, which may be related to the action mechanism of the complex. The complex CBP-01 may induce DNA damage leading cells to accumulate at G0/G1 checkpoint where, apparently, cells that are not able to recover from the damage are driven to cell death. Evidence has shown that cell death is initiated by autophagy dysfunction, culminating in apoptosis induction. The search for new metal-based drugs is focused on overcoming the drawbacks of already used agents such as acquired resistance and non-specificity; thus, the results obtained with CBP-01 show promising effects on cancer cells.
    Keywords:  Apoptosis; Cell cycle; Copper complex; Cytotoxicity; ROS
  37. Planta Med. 2019 Feb 21.
    Zhang BB, Hu XL, Wang YY, Li JY, Pham TA, Wang H.
      Dammarane-type saponins, the main active ingredients of Panax notoginseng, have substantial neuroprotective effects in different animal models of neurodegenerative diseases. However, because these compounds have different structures, the level of protection provided by individual compounds varies, and highly active compounds can be selected based on structure-activity relationships. Glutamate is a major excitatory neurotransmitter that plays an important role in synaptic response development. However, excessive extracellular glutamate levels lead to neuronal dysfunctions in the central nervous system. Herein, we investigated the neuroprotective effects of nine saponins (compounds 1:  - 9: ) on glutamate-treated PC12 cells in the concentration range of 0.1 - 10 µM. The MTT assay revealed that these compounds increased cell viability to 65.6, 69.8, 76.9, 91.7, 74.4, 63.3, 59.9, 64.7, and 59.9%, respectively, compared with the glutamate-treated cells (44.6%). Protopanaxatriol (compound 4: ) was the most neuroprotective compound, and subsequent experiments revealed that pretreatment with compound 4: significantly reverses mitochondrial membrane potential collapse, increases superoxide dismutase activity, and decreases lactate dehydrogenase leakage, malondiadehyde levels, reactive oxygen species generation, and cell apoptosis. Compound 4: also decreased the Bax/Bcl-2 ratio, cleaved caspase-3, N-methyl-D-aspartic receptor 1, and Ca2+-/calmodulin-dependent protein kinase II expression, and inhibited glutamate-induced cytochrome C release and phosphorylation of apoptosis signal-regulating kinase 1, c-Jun N-terminal kinase, and p38. Overall, the results indicate that protopanaxatriol has significant neuroprotective effects, and might be a promising neuroprotective agent for preventing and treating neurodegenerative diseases.
  38. Int J Biol Macromol. 2019 Feb 15. pii: S0141-8130(18)36317-7. [Epub ahead of print]
    Li W, Cai ZN, Mehmood S, Liang LL, Liu Y, Zhang HY, Chen Y, Lu YM.
      Fine particulate matter (PM2.5) exposure could cause many acute and chronic respiratory diseases. In this study the protective effects of polysaccharide from Morchella esculenta (FMP-1) and its derivatives against PM2.5-induced inflammation were evaluated. By flow cytometry and ELISA analysis, sulfated polysaccharide SFMP-1 showed the best protective effect in reducing PM2.5-induced cell death, cell apoptosis and production of tumor necrosis factor-alpha (TNF-α) and interleukin-1 beta (IL-1β), which was accompanied by a diminished level in reactive oxygen species (ROS) formation caused by PM2.5 in rat alveolar macrophage NR8383 cells. Furthermore, the mechanism was studied by immunofluorescence, qRT-PCR and western blotting. SFMP-1 could down-regulate the expression of inducible NO synthesis (iNOS) and cyclooxygenase-2 (COX-2) at both mRNA and protein levels in PM2.5-treated cells. The PM2.5-induced phosphorylation of nuclear factor-kappa B (NF-κB) was also reduced through suppressing nuclear translation of the NF-κB and inhibiting the degradation and phosphorylation of IκBα. These results indicated that SFMP-1 could protect NR8383 cells from PM2.5-induced inflammation by inhibiting NF-κB activation.
    Keywords:  Inflammation; Modification; NR8383 cells; PM(2.5); Polysaccharide
  39. Sci Rep. 2019 Feb 22. 9(1): 2608
    Kim JE, Bae SY, Ahn SY, Kwon YJ, Ko GJ.
      Radiocontrast-induced nephropathy (CIN) is the third most common cause of acute renal failure. The pathophysiology of CIN is related to tubular injury caused by oxidative stress, and nuclear factor erythroid-2-related factor 2 (Nrf2) is critical in coordinating intracellular antioxidative processes. We thus investigated the role of Nrf2 in CIN. CIN was established in mice and in NRK-52E cells via iohexol administration according to the protocols of previous studies. To determine the role of Nrf2 in CIN, Nrf2 expression was reduced in vivo using Nrf2 knockout (KO) mice (B6.129 × 1-Nfe2 l2tm1Ywk/J) and in vitro with siRNA treatment targeting Nrf2. Increased Nrf2 expression was observed after iohexol treatment both in vivo and in vitro. Serum creatinine at 24 h after iohexol injection was significantly higher in KO mice than in wild-type (WT) mice. Histologic examination showed that iohexol-induced tubular vacuolization and structural disruption were aggravated in Nrf2 KO mice. Significant increases in apoptosis and F4/80(+) inflammatory cell infiltration were demonstrated in KO mice compared to WT mice. In addition, the increase in reactive oxygen species after iohexol treatment was augmented by Nrf2 inhibition both in vivo and in vitro. Nrf2 may be implicated in the pathogenesis of CIN via the modulation of antioxidant, anti-apoptotic, and anti-inflammatory processes.
  40. Chem Sci. 2019 Jan 21. 10(3): 879-883
    Zhang W, Zhang J, Li P, Liu J, Su D, Tang B.
      Hepatic ischaemia-reperfusion (IR) injury is mainly attributed to a burst of reactive oxygen species (ROS) that attack biological macromolecules and lead to cell death. The superoxide anion (O2˙-) is the first ROS to be generated and triggers the production of other ROS; thus, explorations of the role of O2˙- in the IR process are meaningful. Meanwhile, the Golgi apparatus generates O2˙- via Golgi-associated proteins, which might play an essential role in IR injury. However, the molecular mechanism by which O2˙- from the Golgi apparatus regulates hepatic IR injury is unclear. Therefore, to solve this problem, a two-photon (TP) excited fluorescence probe (CCA) was designed and prepared for the reversible detection of O2˙- in the Golgi apparatus. With the assistance of TP fluorescence microscopy, we observed a substantial increase in the levels of O2˙- in the Golgi apparatus of an IR mouse liver for the first time, as well as increased caspase-2 activity and apoptosis. Furthermore, we found that the tumour necrosis factor (TNF-α) functions as a positive mediator of O2˙- generation. Based on these data, we identified the potential signalling pathway in the Golgi that mediates O2˙- fluctuations in IR mice and revealed the related molecular mechanisms; we also provide a new target for treating IR injury.
  41. PLoS One. 2019 ;14(2): e0212782
    Lage R, Cebro-Márquez M, Rodríguez-Mañero M, González-Juanatey JR, Moscoso I.
      BACKGROUND: Association between obesity and cardiovascular diseases is well known, however increased susceptibility of obese patients to develop several cancer types is not so commonly known. Current data suggest that poorer overall survival in cancer patients might be associated to non-cancer-related causes such as higher risk of cardiotoxicity in obese patients treated with chemotherapeutic agents. Omentin, a novel adipokine decreased in obesity, is actually in the spotlight due to its favourable effects on inflammation, glucose homeostasis and cardiovascular diseases. Also, recent data showed that in vitro anthracycline-induced cardiomyocyte apoptosis is counteracted by omentin suggesting its cardioprotective role.OBJECTIVE: Our aim was to evaluate omentin effects against docetaxel toxicity.
    RESULTS: Our data indicate that omentin inhibits docetaxel-induced viability loss and that increased viability is associated to decreased caspase-3 expression and cell death. Although omentin reduces NOX4 expression, it failed to reduce docetaxel-induced reactive oxygen species production. Our results indicate that omentin decreases docetaxel-induced endoplasmic reticulum stress, suggesting that cardioprotective role might be associated to ERS inhibition.
    CONCLUSION: These data suggest that omentin treatment may contribute to decrease susceptibility to DTX-induced cardiotoxicity.
  42. J Anim Sci Biotechnol. 2019 ;10 14
    Durfey CL, Swistek SE, Liao SF, Crenshaw MA, Clemente HJ, Thirumalai RVKG, Steadman CS, Ryan PL, Willard ST, Feugang JM.
      Background: Advances in nanotechnology have permitted molecular-based targeting of cells through safe and biocompatible magnetic nanoparticles (MNP). Their use to detect and remove damaged spermatozoa from semen doses could be of great interest. Here, MNP were synthesized and tested for their ability to target apoptotic (annexin V) and acrosome-reacted (lectin) boar spermatozoa, for high-throughout retrieval in a magnetic field (nanoselection). The potential impacts of nanoselection on sperm functions and performance of offspring sired by sperm subjected to nanoselection were determined. Fresh harvested and extended boar semen was mixed with various amounts (0, 87.5, and 175 μg) of MNP-conjugates (Annexin V-MNP or Lectin-MNP) and incubated (10 to 15 min) for 37 °C in Exp. 1. In Exp. 2, extended semen was mixed with optimal concentrations of MNP-conjugates and incubated (0, 30, 90, or 120 min). In Exp. 3, the synergistic effects of both MNP-conjugates (87.5 μg - 30 min) on spermatozoa was evaluated, followed by sperm fertility assessments through pregnancy of inseminated gilts and performance of neonatal offspring. Sperm motion, viability, and morphology characteristics were evaluated in all experiments.Results: Transmission electron microscopy, atomic force microscopy, and hyperspectral imaging techniques were used to confirm attachment of MNP-conjugates to damaged spermatozoa. The motility of nanoselected spermatozoa was improved (P < 0.05). The viability of boar sperm, as assessed by the abundance of reactive oxygen species and the integrity of the acrosome, plasma membrane, and mitochondrial membrane was not different between nanoselected and control spermatozoa. The fertility of gilts inseminated with control or nanoselected spermatozoa, as well as growth and health of their offspring were not different between (P > 0.05).
    Conclusions: The findings revealed the benefit of magnetic nanoselection for high-throughput targeting of damaged sperm, for removal and rapid and effortless enrichment of semen doses with highly motile, viable, and fertile spermatozoa. Therefore, magnetic nanoselection for removal of abnormal spermatozoa from semen is a promising tool for improving fertility of males, particularly during periods, such as heat stress during the summer months.
    Keywords:  Acrosome reaction; Apoptosis; Artificial insemination; Boar; Iron oxide nanoparticles; Nanopurification; Nanoselection; Nanotechnology; Reproduction; Swine
  43. Free Radic Biol Med. 2019 Feb 15. pii: S0891-5849(18)32442-0. [Epub ahead of print]
    Sb H, X J, Qh Y, Xr Z, Bb Z, Kh W, Xy S, Yt C, Xr R, Jf M, G W, Yh P.
      Oxidative stress (OS) plays crucial roles in triethylene glycol dimethacrylate (TEGDMA, a major component in dental resin)-induced apoptosis of dental pulp cells. Mitochondria are important target organelles for regulating the balance of OS, meanwhile, imbalance of the mitochondrial dynamic associated with mitochondrial dysfunction is one major molecular mechanism for oxidative damages. However, whether these mitochondrial dependent pathways were involved in the apoptosis of dental pulp cells induced by TDGDMA remains unclarified. We demonstrated that TEGDMA decreased viability and induced apoptosis of mouse preodontoblasts (mDPC6T cell line) in a time- and dose-dependent manner. Furthermore, TEGDMA elevated the mitochondrial OS status and induced mitochondrial dysfunction, as reflected by the significant decrease of mitochondrial membrane potential, ATP production, the activity of Complex III and citrate synthase. In this process, we detected a dramatically impaired mitochondrial dynamic that was reflected by significantly enhanced mitochondrial fragmentation. Consistently, we also found a significant enhancement of the key upstream regulators for mitochondrial fission, such as short form of optic atrophy 1, dynamic related protein 1 oligomer and Fission 1. The respective inhibition of mitochondrial OS or mitochondrial fission could mutually attenuate each other, thereby significantly preventing both mitochondrial dysfunction and cell apoptosis. In conclusion, TEGDMA-induced preodontoblasts apoptosis was mediated by the vicious circle between mitochondrial OS and dynamic abnormality, which represented a new target to prevent TEGDMA-induced dental pulp cells apoptosis.
    Keywords:  Apoptosis; Dental pulp cell; Mitochondrial dynamic; Mitochondrial dysfunction; Oxidative stress; TEGDMA
  44. Chem Biol Interact. 2019 Feb 15. pii: S0009-2797(18)31495-9. [Epub ahead of print]
    Khan S, Arif SH, Naseem I.
      Riboflavin (Rf) or vitamin B₂ is a known photosensitizer whose photophysical and photochemical properties are well established. Aminophylline (Am) is a phosphodiesterase inhibitor and is currently used as a bronchodilator. Although there are several reports of haemolytic and proteolytic interaction of photoilluminated riboflavin with aminophylline, the cytotoxicity of this system against malignant tissue is not well defined and fully unravelled. Here, we are evaluating anticancer activity of this system against B(a)P induced lung carcinoma in swiss albino mice. We observed marked increment in the level of cellular redox scavengers as well as oxidative stress markers. A significant DNA damage was observed using comet assay. Histopathological studies further confirmed induction of apoptosis in lung tissues of Am-Rf treated animals. Scanning electron microscopy revealed altered surface morphology of the malignant tissue, which characteristically improved in the treatment group. Since malignancy is characterised by compromised redox status, therefore, further increment in ROS due to the action of this system derives cellular system towards extensive macromolecular damage and consequent ROS mediated apoptosis. We anticipate the usage of this system in developing efficient photodynamic therapy against lung cancer that can be clinically realised.
    Keywords:  Aminophylline; Apoptosis; DNA damage; Photodynamic therapy; ROS; Riboflavin
  45. FASEB J. 2019 Feb 20. fj201801921R
    Wang M, Hu J, Yan L, Yang Y, He M, Wu M, Li Q, Gong W, Yang Y, Wang Y, Handy DE, Lu B, Hao C, Wang Q, Li Y, Hu R, Stanton RC, Zhang Z.
      Oxidative stress contributes substantially to podocyte injury, which plays an important role in the development of diabetic kidney disease. The mechanism of hyperglycemia-induced oxidative stress in podocytes is not fully understood. Glucose-6-phosphate dehydrogenase (G6PD) is critical in maintaining NADPH, which is an important cofactor for the antioxidant system. Here, we hypothesized that high glucose induced ubiquitination and degradation of G6PD, which injured podocytes by reactive oxygen species (ROS) accumulation. We found that G6PD protein expression was decreased in kidneys of both diabetic patients and diabetic rodents. G6PD activity was also reduced in diabetic mice. Overexpressing G6PD reversed redox imbalance and podocyte apoptosis induced by high glucose and palmitate. Inhibition of G6PD with small interfering RNA induced podocyte apoptosis. In kidneys of G6PD-deficient mice, podocyte apoptosis was significantly increased. Interestingly, high glucose had no effect on G6PD mRNA expression. Decreased G6PD protein expression was mediated by the ubiquitin proteasome pathway. We found that the von Hippel-Lindau (VHL) protein, an E3 ubiquitin ligase subunit, directly bound to G6PD and degraded G6PD through ubiquitylating G6PD on K366 and K403. In summary, our data suggest that high glucose induces ubiquitination of G6PD by VHL E3 ubiquitin ligase, which leads to ROS accumulation and podocyte injury.-Wang, M., Hu, J., Yan, L., Yang, Y., He, M., Wu, M., Li, Q., Gong, W., Yang, Y., Wang, Y., Handy, D. E., Lu, B., Hao, C., Wang, Q., Li, Y., Hu, R., Stanton, R. C., Zhang, Z. High glucose-induced ubiquitination of G6PD leads to the injury of podocytes.
    Keywords:  G6PD; diabetic kidney disease; reactive oxygen species; ubiquitin proteasome pathway; von Hippel-Lindau
  46. Aging (Albany NY). 2019 Feb 14. 11(3): 1030-1044
    Wu M, Ma L, Xue L, Ye W, Lu Z, Li X, Jin Y, Qin X, Chen D, Tang W, Chen Y, Hong Z, Zhang J, Luo A, Wang S.
      Chemotherapy-induced ovarian aging not only increases the risk for early menopause-related complications but also results in infertility in young female cancer survivors. Oogonial stem cells have the ability to generate new oocytes and thus provide new opportunities for treating ovarian aging and female infertility. Resveratrol (3,5,4'-trihydroxy-trans-stilbene) is a natural phenol derived from plants, that has been shown to have positive effects on longevity and redox flow in lipid metabolism and a preventive function against certain tumors. To evaluate whether resveratrol could promote the repair of oogonial stem cells damage in a busulfan/cyclophosphamide (Bu/Cy)-induced accelerated ovarian aging model, female mice were administered 30 and 100 mg/kg/d resveratrol through a gavage for 2 weeks. We demonstrated that resveratrol (30 mg/kg/d) relieved oogonial stem cells loss and showed an attenuating effect on Bu/Cy-induced oxidative apoptosis in mouse ovaries, which may be attributed to the attenuation of oxidative levels in ovaries. Additionally, we also showed that Res exerted a dose-dependent effect on oogonial stem cells and attenuated H2O2-induced cytotoxicity and oxidative stress injury by activating Nrf2 in vitro. Therefore, resveratrol could be of a potential therapeutic drug used to prevent chemotherapy-induced ovarian aging.
    Keywords:  apoptosis; chemotherapy; oogonial stem cell; ovarian aging; resveratrol
  47. Nutr Neurosci. 2019 Feb 21. 1-16
    Mohammadzadeh L, Abnous K, Razavi BM, Hosseinzadeh H.
      Organophosphorus compounds are widely used in agriculture. Epidemiological studies propose that pesticide exposure is a risk factor for Alzheimer's disease (AD), but the mechanisms are unclear. Here, we investigated the impact of malathion exposure on the cognitive ability and the underlying mechanisms in rats. Moreover, we studied whether crocin reduced malathion-induced cognitive and memory loss in rats. Malathion (100 mg/kg) and crocin (10, 20 and 40 m/kg) were administered into the rats once a day for 14 days via i.p. Also vitamin E was used as positive control. Malathion exhibited spatial memory deficits as assessed by Morris water maze (MWM). Malathion increased the latency to reach the platform and decreased time spent and swimming distance of animals in target quadrant in probe trial. These effects were protected by crocin. Malathion exposure induced spatial learning and memory deficits with a simultaneous decrease of PSD93 and TAU hyperphosphorylation at multiple AD-related phosphorylation sites with activation of glycogen synthase kinase-3β (GSK-3β) and inhibition of protein phosphatase-2A (PP2A). Additionally, the elevation of malondialdehyde (MDA), TNF α and IL-6 levels, amelioration of reduced glutathione (GSH) in the hippocampus and reduction of plasma acetylcholinesterase activity were observed upon administration of the malathion. Also, malathion-induced apoptosis in the hippocampus. Crocin or vitamin E improved memory damages and antagonized the effects of malathion. According to the data of this study, crocin mitigated malathion-induced neurological alterations and cognitive impairment by reducing oxidative stress and inflammation, inhibiting TAU protein hyperphosphorylation and antiapoptotic effects.
    Keywords:  Apoptosis; Crocin; Lipid peroxidation; Malathion; Memory impairment; Morris water maze; TAU
  48. Cell Death Dis. 2019 Feb 20. 10(3): 173
    Lin L, Li X, Pan C, Lin W, Shao R, Liu Y, Zhang J, Luo Y, Qian K, Shi M, Bin J, Liao Y, Liao W.
      For gastric cancer (GC) control, metastasis and chemoresistance are the major challenges, accompanied with various stresses. Ataxin-2-like (ATXN2L) was discovered as a novel regulator of stress granules, yet its function in cancers remained unknown. Hence, we wanted to explore the functions of ATXN2L to see whether it participates in stress-related cancer malignant activities. Clinical follow-up was performed to see the impact of ATXN2L on GC patient survival. As a result, ATXN2L expression was upregulated in GC tissue and indicated adverse prognosis for overall survival and recurrence. In GC cells, ATXN2L expression was knocked down and functional experiments were performed. ATXN2L promoted GC cell migration and invasion via epithelial to mesenchymal transition, yet no influence on proliferation was detected by ATXN2L interference. When adding the chemotherapeutic agent oxaliplatin to induce stress, silencing ATXN2L sensitized GC cells to oxaliplatin. Interestingly, oxaliplatin was found to in turn promote ATXN2L expression and stress granule assembly. Then, two acquired oxaliplatin-resistant strains were generated by long-term oxaliplatin induction. The oxaliplatin-resistant strains presented with elevated ATXN2L levels, while silencing ATXN2L in the strains reversed the oxaliplatin resistance by increasing reactive oxygen species production and apoptosis. These results suggested that ATXN2L was responsible for not only intrinsic but also acquired oxaliplatin chemoresistance. Finally, ATXN2L-related signaling was screened using bioinformatic methods, and epidermal growth factor (EGF) was verified to promote ATXN2L expression via PI3K/Akt signaling activation. Blocking EGFR/ATXN2L signaling reversed GC cell oxaliplatin resistance and inhibited migration. In conclusion, ATXN2L promotes cell invasiveness and oxaliplatin resistance and can be upregulated by EGF via PI3K/Akt signaling. ATXN2L may be an indicator and therapeutic target in GC, especially for oxaliplatin-based chemotherapy.
  49. J Biol Chem. 2019 Feb 21. pii: jbc.RA118.007102. [Epub ahead of print]
    Conteh AM, Reissaus CA, Hernandez-Perez M, Nakshatri S, Anderson RM, Mirmira RG, Tersey SA, Linnemann AK.
      In type 1 diabetes, an autoimmune event increases oxidative stress in islet β cells, giving rise to cellular dysfunction and apoptosis. Lipoxygenases are enzymes that catalyze the oxygenation of polyunsaturated fatty acids that can form lipid metabolites involved in several biological functions including oxidative stress. 12-lipoxygenase and 12/15-lipoxygenase are related but distinct enzymes that are expressed in pancreatic islets, but their relative contributions to oxidative stress in these regions are still being elucidated. In this study, we used mice with global genetic deletion of the genes encoding 12-lipoxygenase (arachidonate 12-lipoxygenase, 12S type [Alox12]) or 12/15-lipoxygenase (Alox15) to compare the influence of each gene deletion on β-cell function and survival in response to the β-cell toxin streptozotocin (STZ). Alox12-/- mice exhibited greater impairment in glucose tolerance following STZ exposure than did wild-type mice (WT), whereas Alox15-/- mice were protected against dysglycemia. These changes were accompanied by evidence for islet oxidative stress in Alox12-/- mice and reduced oxidative stress in Alox15-/- mice, consistent with alterations in the expression of the antioxidant response enzymes in islets from these mice. Additionally, islets from Alox12-/- mice displayed a compensatory increase in Alox15 gene expression, and treatment of these mice with the 12/15-lipoxygenase inhibitor ML-351 rescued the dysglycemic phenotype. Collectively, these results indicate that Alox12 loss activates a compensatory increase in Alox15 that sensitizes β cells to oxidative stress. These findings support the conclusion that Alox15 represents the more relevant target in efforts to protect β cells from oxidative stress.
    Keywords:  diabetes; lipoxygenase pathway; oxidative stress; pancreatic islet; reactive oxygen species (ROS)
  50. Int J Nanomedicine. 2019 ;14 951-962
    Li C, Wang Y, Zhang H, Li M, Zhu Z, Xue Y.
      Background: Gastric cancer is the fourth most common cancer and second leading cause of cancer death worldwide. Cardiospermum halicacabum is used to treat nerve disorders, stiffness, rheumatism, ear ache, snake bite, and so on.Methods: In this study, the reaction parameters were optimized to control the size of the nanoparticle, which was confirmed by transmission electron microscopy. Various characterization techniques such as selected area diffraction pattern, UV-visible spectroscopy, energy-dispersive X-ray analysis, dynamic light scattering, Fourier-transform infrared spectroscopy, and atomic force microscopy were employed to analyze the synthesized AuNPs obtained from C. halicacabum (CH-AuNP) against gastric carcinoma cell line.
    Results: The cytotoxic effect of CH-AuNP against AGS, SNU-5, and SNU-16 cell lines was detected by MTT assay. The induction of apoptosis by CH-AuNP in AGS was analyzed by double staining technique using TUNEL and DAPI staining assays. Further to confirm the molecular mechanism exhibited by CH-AuNP to induce apoptosis, the intracellular ROS level was assessed and immunoblotting was performed to assess the apoptotic signaling molecules that often deregulated in cancerous condition.
    Conclusion: The results clearly prove that CH-AuNP increases ROS and induces apoptosis in AGS, suggesting that CH-AuNP may be an effective anticancer drug with no side effects to treat gastric cancer.
    Keywords:  MMP; ROS; apoptosis; gastric cancer; gold nanoparticle Cardiospermum halicacabum
  51. Mol Metab. 2019 Feb 08. pii: S2212-8778(18)31115-3. [Epub ahead of print]
    Baboota RK, Shinde AB, Lemaire K, Fransen M, Vinckier S, Van Veldhoven PP, Schuit F, Baes M.
      OBJECTIVES: Peroxisomes play a crucial role in lipid and reactive oxygen species metabolism, but their importance for pancreatic β-cell functioning is presently unknown. To examine the contribution of peroxisomal metabolism to β-cell homeostasis in mice, we inactivated PEX5, the import receptor for peroxisomal matrix proteins, in an inducible and β-cell restricted manner (Rip-Pex5-/- mice).METHODS: After tamoxifen-induced recombination of the Pex5 gene at the age of 6 weeks, mice were fed either normal chow or a high-fat diet for 12 weeks and were subsequently phenotyped.
    RESULTS: Increased levels of very long chain fatty acids and reduced levels of plasmalogens in islets confirmed impairment of peroxisomal fatty acid oxidation and ether lipid synthesis, respectively. The Rip-Pex5-/- mice fed on either diet exhibited glucose intolerance associated with impaired insulin secretion. Ultrastructural and biochemical analysis revealed a decrease in the density of mature insulin granules and total pancreatic insulin content, which was further accompanied by mitochondrial disruptions, reduced complex I activity and massive vacuole overload in β-cells. RNAseq analysis suggested that cell death pathways were affected in islets from HFD-fed Rip-Pex5-/- mice. Consistent with this change we observed increased β-cell apoptosis in islets and a decrease in β-cell mass.
    CONCLUSIONS: Our data indicate that normal peroxisome metabolism in β-cells is crucial to preserve their structure and function.
    Keywords:  Apoptosis; Diabetes; High-fat diet; Islet; Peroxisome; β-cell
  52. J Agric Food Chem. 2019 Feb 19.
    Zhuang S, Yu R, Zhong J, Liu P, Liu Z.
      Rheum rhabarbarum has been widely used as a herbal medicine and food in China. The objective of this study was to investigate the cytoprotective action and underlying mechanisms of rhein, one active ingredient isolated from Rheum rhabarbarum, on H2O2-challenged rat small intestine epithelial cells (IEC-6 cells). H2O2-challenged IEC-6 cells were incubated in the pretreatment with or without rhein or LY294002, a PI3K/Akt inhibitor. The cell viability, apoptosis, intracellular ROS and antioxidants were measured. The expressions of heme oxygenase-1 (HO-1), nuclear NF-erythroid 2-related factor (Nrf2), Akt and p-Akt were evaluated by western blotting. Meanwhile, LY294002 was also used to investigate the role of PI3K/Akt in rhein-induced cytoprotective role. The results showed that pretreatment of rhein could reverse the inhibition of cell viability (95.71±6.80% under 2 μM rhein and 92.61±3.76% under 4 μM rhein compared with 78.91±1.72% under stimulation with H2O2 alone) and suppress the apoptosis (17.12±1.65% under 2 μM rhein and 16.43±1.14% under 4 μM rhein compared with 22.58±0.61% under stimulation with H2O2 alone), caspase-3 activity (115.20±7.47% under 2 μM rhein and 128.98±26.98% under 4 μM rhein compared with 157.04±15.59% under stimulation with H2O2 alone) and intracellular ROS (157.79±31.98% under 2 μM rhein and 141.83±17.37% under 4 μM rhein compared with 196.52±21.26% under stimulation with H2O2 alone) induced by H2O2. Rhein also supported SOD activity (87.90±9.80% under 2 μM rhein and 125.12±14.09% under 4 μM rhein compared with 66.25±12.98% under stimulation with H2O2 alone), catalase activity (90.71±15.37% under 2 μM rhein and 82.66±19.06% under 4 μM rhein compared with 57.15±8.90% under stimulation with H2O2 alone), GST activity (83.31±13.45% under 2 μM rhein and 111.18±3.07% under 4 μM rhein compared with 48.67±16.02% under stimulation with H2O2 alone) and GSH content (122.05±10.05% under 2 μM rhein and 94.42±16.94% under 4 μM rhein compared with 70.71±11.78% under stimulation with H2O2 alone). Furthermore, rhein induced the protein expression of HO-1 (1.64±0.05-fold under 2 μM rhein and 2.07±0.27-fold under 4 μM rhein compared with 1.30±0.16-fold with H2O2 challenge alone) together with its upstream mediator Nrf2 (1.64±0.05-fold under 2 μM rhein and 2.07±0.27-fold under 4 μM rhein compared with 1.30±0.16-fold under stimulation with H2O2 alone), and activated the phosphorylation of Akt (1.01±0.09-fold under 2 μM rhein and 0.89±0.06-fold under 4 μM rhein compared with 0.75±0.04-fold with H2O2 challenge alone) in IEC-6 cells. LY294002 inhibited increased cell viability (90.57±2.27% compared with 97.71±2.88% under 2 μM rhein and 88.55±2.41% compared with 97.09±3.97% under 4 μM rhein), up-regulated lowered apoptotic rate (29.27±2.95% compared with 17.12±1.65% under 2 μM rhein and 20.98±0.55% compared with 16.43±1.14% under 4 μM rhein) and enhanced the weakened ROS levels (205.67±30.03% compared with 156.79±31.98% under 2 μM rhein and 173.20±0.55% compared with 141.83±17.37% under 4 μM rhein). Although the inhibition of PI3K/Akt did not inhibit the Nrf2 nuclear level (1.87±0.12-fold compared with 2.01±0.18-fold) under 4 μM rhein concentration, LY294002 inhibited Nrf2 nuclear level (1.80±0.29-fold compared with 2.94±0.47-fold) under 2 μM rhein concentration and blocked HO-1 expression (1.87±0.21-fold compared with 2.19±0.10-fold under 2 μM rhein and 2.55±0.30-fold compared with 3.01±0.10-fold under 4 μM rhein). These data demonstrated that rhein protected IEC-6 cells against oxidative damage partly via PI3K/Akt and Nrf2/HO-1 pathways.
  53. Bioorg Chem. 2019 Feb 08. pii: S0045-2068(18)31019-8. [Epub ahead of print]85 455-468
    De Grandis RA, Santos PWDSD, Oliveira KM, Machado ART, Aissa AF, Batista AA, Antunes LMG, Pavan FR.
      This study describes a series of newly synthesized phosphine/diimine ruthenium complexes containing the lawsone as bioligand with enhanced cytotoxicity against different cancer cells, and apoptosis induction in prostatic cancer cells DU-145. The complexes [Ru(law)(N-N)2]PF6 where N-N is 2,2'-bipyridine (1) or 1,10-phenanthroline (2) and [Ru(law)(dppm)(N-N)]PF6, where dppm means bis(diphenylphosphino)methane, N-N is 2,2'-bipyridine (3) or 1,10-phenanthroline (4), and law is lawsone, were synthesized and fully characterized by elemental analysis, molar conductivity, NMR, UV-vis, IR spectroscopies and cyclic voltammetry. The interaction of the complexes (1-4) with DNA was evaluated by circular dichroism, gel electrophoresis, and fluorescence, and the complexes presented interactions by the minor grooves DNA. The phosphinic series of complexes exhibited a remarkably broad spectrum of anticancer activity with approximately 34-fold higher than cisplatin and 5-fold higher than doxorubicin, inhibiting the growth of 3D tumor spheroids and the ability to retain the colony survival of DU-145 cells. Also, the complex (4) inhibits DU-145 cell adhesion and migration potential indicating antimetastatic properties. The mechanism of its anticancer activity was found to be related to increased reactive oxygen species (ROS) generation, increased the BAX/BCL-2 ratio and subsequent apoptosis induction. Overall, these findings suggested that the complex (4) could be a promising candidate for further evaluation as a chemotherapeutic agent in the prostate cancer treatment.
    Keywords:  1,4-naphthoquinone; 3D-cell culture; Apoptosis; BAX/BCL-2; Cancer; ROS