bims-aporos Biomed news
on Apoptosis and reactive oxygen species
Issue of 2019‒02‒10
sixty-one papers selected by
Gavin McStay
Staffordshire University


  1. Environ Sci Pollut Res Int. 2019 Feb 02.
    Li S, Xu Z, Xia J, Qin G, Sang N.
      Epidemiological evidence suggests that the incidence and mortality of cardiovascular diseases are closely related to sulfur dioxide (SO2). In the present study, H9C2 cells were incubated with 100 μM NaHSO3 with or without pretreatment of an antioxidant, N-acetyl-L-cysteine (NAC). The changes of apoptosis rate, mitochondrial membrane potential (MMP), ATP content, caspase-3 activity, and reactive oxygen species (ROS) were detected. Rats were inhaled 7 mg/m3 SO2 and/or intraperitoneal injected with 50 mg/kg (bw) of NAC for 30 days. RT-PCR and Western blot were used to detect the mRNA and protein levels of apoptosis-related genes. We found that the apoptosis of H9C2 cells was induced by NaHSO3, which decreased the content of MMP and ATP, and induced the expression of caspase-3. NAC can inhibit the apoptosis induced by NaHSO3 treatment. SO2 and NaHSO3 decreased the expression of Bcl-2 and the ratio of Bcl-2/Bax, increased the expression of Bax and P53 accumulation and phosphorylation, and activated caspase-9 and caspase-3. Whereas NAC can reduce the changes of apoptosis-related proteins in rat heart. Our results suggest that SO2 induces ROS-mediated P53 and caspase-dependent mitochondrial signaling pathways in H9C2 cells and rat hearts. Antioxidant therapy can reduce the adverse reactions of SO2 and lead to a decline in the cardiovascular disease induced by SO2.
    Keywords:  Antioxidant; Apoptosis; Hearts; Reactive oxygen species; Sulfur dioxide
    DOI:  https://doi.org/10.1007/s11356-019-04319-7
  2. J Cell Physiol. 2019 Feb 03.
    Wang L, Yan W, Wang J.
      MEKK3 is a member of MAP3K, which plays a pivotal role in cardiac diseases. In this study, we aimed to investigate the effects and potential mechanisms of MEKK3 on hypoxia/reoxygenation (H/R) injury of cardiomyocytes. After exposing H9C2 cells to H/R insult, real-time polymerase chain reaction and western blot analysis showed that MEKK3 was highly expressed. Cell viability, cell apoptosis, caspase 3/7 activity, and cleaved-caspase 3 expression were tested using a CCK-8 assay, Cell Death Detection PLUS ELISA, Caspase-Glo 3/7 Assay Kit and western blot analysis, respectively. Mitochondrial membrane potential, cytochrome C expression, adenosine triphosphate (ATP), and reactive oxygen species also were measured using JC-1 staining, western blot analysis, an ATP Assay Kit, and DCFH 2 -DA staining, respectively. The messenger RNA (mRNA) levels and secretions of TNF-α, IL-6, and IL-1β were evaluated. The results revealed that MEKK3 silencing promoted cell survival and attenuated lactate dehydrogenase leakage, cell apoptosis, caspase 3/7 activity, and the protein level of cleaved-caspase 3. Moreover, knockdown of MEKK3 blocked mitochondrial impairment by inhibiting the loss of mitochondrial membrane potential and cytochrome C expression as well as promoting ATP synthesis. MEKK3 deficiency led to a decrease in reactive oxygen species and malondialdehyde (MDA) generation and an increase in superoxide dismutase (SOD) activity. Deletion of MEKK3 led to reduced inflammatory cytokines in mRNA level and secretion. MEKK3 suppression activated the sonic hedgehog (Shh) signaling pathway in H9C2 cells. After blocking the Shh signaling pathway with a specific inhibitor, cyclopamine, the cardioprotective functions of MEKK3 downregulation were partly abolished. In conclusion, downregulation of MEKK3 prevented apoptosis and inflammation in H9C2 cells via the Shh signaling pathway.
    Keywords:  H/R; MEKK3; Shh signaling pathway; apoptosis; inflammation
    DOI:  https://doi.org/10.1002/jcp.28162
  3. Oxid Med Cell Longev. 2019 ;2019 2593742
    Cui J, Liu Y, Chang X, Gou W, Zhou X, Liu Z, Li Z, Wu Y, Zuo D.
      Excessive drinking can damage brain tissue and cause cognitive dysfunction. Studies have found that the early stage of neurodegenerative disease is closely related to heavy drinking. Acetaldehyde (ADE) is the main toxic metabolite of alcohol. However, the exact mechanisms of ADE-induced neurotoxicity are not fully clear. In this article, we studied the cytotoxic effect of ADE in HT22 cells and primary cultured cortical neuronal cells. We found that ADE exhibited cytotoxicities against HT22 cells and primary cultured cortical neuronal cells in dose-dependent manners. Furthermore, ADE induced apoptosis of HT22 cells by upregulating the expression of caspase family proapoptotic proteins. Moreover, ADE treatment could significantly increase the intracellular Ca2+ and reactive oxygen species (ROS) levels and activate endoplasmic reticulum stress (ERS) in HT22 cells. ADE upregulated ERS-related CHOP expression dose-dependently in primary cultured cortical neuronal cells. In addition, inhibition of ROS with antioxidant N-acetyl-L-cysteine (NAC) reduced the accumulation of ROS and reversed ADE-induced increase of ERS-related protein and apoptosis-related protein levels. Mitigation of ERS with ERS inhibitor 4-PBA obviously suppressed ADE-induced apoptosis and the expression of ERS-related proteins. Therefore, ADE induces neurotoxicity of HT22 cells via oxidative stress- and Ca2+ imbalance-mediated ERS.
    DOI:  https://doi.org/10.1155/2019/2593742
  4. Vet Immunol Immunopathol. 2019 Feb;pii: S0165-2427(18)30389-1. [Epub ahead of print]208 16-24
    Maina T, Prysliak T, Perez-Casal J.
      Bacterial pathogens have evolved to manipulate host cell death and survival pathways for their intracellular persistence. Understanding the ability of a bacterium to induce or inhibit cell death is essential for elucidating the disease pathogenesis and suggesting potential therapeutic options to manage the infection. In recent years, apoptosis inhibition by different bacteria has been suggested as a mechanism of survival by allowing the pathogen to replicate and disseminate in the host. Mycoplasma bovis has evolved mechanisms to invade and modulate apoptosis of bovine peripheral blood mononuclear cells (PBMC), red blood cells (RBCs), primary macrophages and monocytes. To date, these mechanisms are poorly understood. Using apoptosis assays such as Annexin V binding, caspases activity, reactive oxygen species production, DNA fragmentation and differential gene expression we set out to determine how M. bovis modulates macrophage survival. Using the BoMac cell line, we report a significant reduction in STS-induced apoptosis through caspase dependent manner. Besides activating the NF-kβ pathway and inhibiting caspases 3, 6 and 9, M. bovis strain Mb1 also inhibits production of reactive oxygen species and DNA fragmentation of the host cell. We also report a significant up-regulation of the anti-apoptotic genes Bcl-2 and Bcl-XL upon infection. Our results indicate that M. bovis strain Mb1 inhibits the intrinsic pathway of apoptosis and up-regulate survival genes in BoMac cells.
    Keywords:  Apoptosis; BoMac cell line; Cattle; Cytochrome C; DNA fragmentation; Mycoplasma bovis
    DOI:  https://doi.org/10.1016/j.vetimm.2018.12.004
  5. Biochem Biophys Res Commun. 2019 Feb 02. pii: S0006-291X(19)30154-8. [Epub ahead of print]
    Yao J, Li Z, Fu Y, Wu R, Wang Y, Liu C, Yang L, Zhang H.
      In this study, we evaluated the expression and function of chemerin and CMKLR1 in the ovaries and granulosa cells of high-fat diet-induced obese (OB) mice. In vivo, chemerin/CMKLR1 system was upregulated in the serum, ovaries, and granulosa cells of OB mice compared with those in control mice. Apoptotic ovarian follicles, oxidative stress, and apoptosis biomarkers were also increased in the ovaries of OB mice. In vitro, mouse granulosa cells (mGCs) were cultured and treated with different concentrations of chemerin to investigate the effects of chemerin on viability, reactive oxygen species (ROS), and apoptosis and on the phosphorylation of AKT, AMP-activated protein kinase α (AMPKα), and nuclear factor-κB p65. Chemerin suppressed mGC viability with or without gonadotrophin and induced ROS accumulation and apoptosis in mGCs. Moreover, AMPKα and p65 were activated by chemerin, whereas AKT was suppressed. These changes in phosphorylation were blocked with CMKLR1 knockdown. Our findings showed that chemerin contributed to ROS accumulation and apoptotic cell death through three signaling pathways, suggesting that upregulation of chemerin and CMKLR1 may explain the imbalance of oxidative stress and apoptosis in the ovaries of OB mice.
    Keywords:  Apoptosis; Chemerin; Granulosa cells; Obesity; Oxidative stress
    DOI:  https://doi.org/10.1016/j.bbrc.2019.01.125
  6. Med Sci Monit. 2019 Feb 05. 25 1009-1016
    Zhou Z, Chen Y, Ni W, Liu T.
      BACKGROUND Endoplasmic reticulum stress (ERS) is part of the cardiovascular pathological processes, including atherosclerosis. Nuclear factor IA (NFIA) influences atherosclerosis development; however, its effects on ERS remain unknown. This study investigated the effect of NFIA on oxidized low-density lipoprotein (ox-LDL)-induced ERS and apoptosis in endothelial cells. MATERIAL AND METHODS Ox-LDL was used to induce lipotoxicity in human umbilical vein endothelial cells (HUVECs) to establish an in vitro oxidative injury model transfected with pcDNA3.0-NFIA. The cytotoxic response was detected using an assay to determine the release of lactate dehydrogenase (LDH). Morphological changes in cell apoptosis were detected using Hoechst 33258 staining. The proportion of apoptotic cells, releases of reactive oxygen species (ROS), and mitochondrial membrane potential (∆ψm) were determined using flow cytometry. The expression levels of apoptosis- and ERS-related molecules were detected through Western blotting. RESULTS NFIA expression was downregulated in the in vitro oxidative cell-injury model. Exposure of HUVECs to ox-LDL resulted in a significant increase in apoptosis, decrease in ROS levels, and loss of ∆ψm. Overexpression of NFIA remarkably inhibited ERS and mitochondrial-mediated apoptosis induced by ox-LDL in HUVECs by reversing the effect of ox-LDL on the expression of JNK1, p-JNK1, CHOP, Cyt C, and Bax. CONCLUSIONS These results demonstrated that NFIA might have beneficial effects in the prevention of ox-LDL-induced ERS and apoptosis in vascular endothelial cells. This study provided new insights into the mechanism of atherosclerosis.
    DOI:  https://doi.org/10.12659/MSM.912132
  7. J Toxicol Sci. 2019 ;44(2): 73-81
    Hirata T, Cho YM, Suzuki I, Toyoda T, Akagi JI, Nakamura Y, Numazawa S, Ogawa K.
      To investigate the chemopreventive mechanisms of 4-Methylthio-3-butenyl isothiocyanate (MTBITC), we analyzed cell viability, cell cycle distribution, and expression levels for cell cycle and apoptosis-related proteins in MTBITC-treated malignant esophageal KYSE510 cells, with and without the reactive oxygen species (ROS) scavenger N-acethyl-L-Cysteine (NAC). MTBITC dose-dependently reduced cell viability and Bcl2 protein expression, while it induced cleavages of caspase-3, caspase-9, and PARP-1, suggesting that reduced cell viability occurred through the mitochondrial apoptotic pathway in KYSE510 cells. In cell cycle distribution analysis, MTBITC (20-40 µM) induced cell cycle arrest at G2/M phase. Furthermore, MTBITC induced Chk1 and Akt phosphorylations and decreased p27 protein expression. Both apoptotic- and cell cycle-related changes induced by MTBITC treatment were abolished by NAC. These results suggest that MTBITC has chemopreventive potential for esophageal carcinogenesis by elimination of cancer cells via induction of mitochondrial apoptotic cell death, G2/M cell cycle arrest, and ROS production.
    Keywords:  4-Methylthio-3-butenyl isothiocyanate (MTBITC); Apoptosis; Chemoprevention; Esophageal cancer cells; G2/M cell cycle arrest
    DOI:  https://doi.org/10.2131/jts.44.73
  8. Cardiovasc Eng Technol. 2019 Feb 06.
    Ma M, Ma X, Cui J, Guo Y, Tang X, Chen C, Zhu Y, Cui C, Wang G.
      PURPOSE: This study aims to investigate the effects and the molecular mechanism of cyclosporin A (CsA) against oxidative stress injury in cultured neonatal rat cardiomyocytes.METHODS: Bax/Bcl-2, cl-casp-9/casp-9, cl-casp-3/casp-3, and iNOS/β-actin ratios and p-IκB and IκB levels were analyzed by western blot. IL-1β and TNF-α levels were analyzed by ELISA.
    RESULTS: CsA effectively improved the cell viability and reduced the extracellular lactate dehydrogenase release in cardiomyocytes after H2O2-induced oxidative damage. CsA significantly increased the superoxide dismutase activity, glutathione production, and catalase activity but decreased the malonaldehyde level. CsA treatment considerably reduced the H2O2-induced intracellular generation of reactive oxygen species, mitochondrial dysfunction, and release of cytochrome c. CsA could act against H2O2-induced ATP reduction, TCA cycle enzymes, mitochondrial complex I enzyme, and complex V enzyme in cardiomyocytes. CsA significantly decreased the Bax/Bcl-2 ratio, cl-casp-9/casp-9, and cl-casp-3/casp-3 in a concentration-dependent manner. CsA also remarkably reduced the cleaved PARP level and DNA fragmentation. NF-κB was closely related to oxidative stress injury. CsA inhibited NF-κB activation, thereby preventing the upregulation of IL-1β, TNF-α, iNOS, and intracellular NO release.
    CONCLUSIONS: CsA protected cardiomyocytes against H2O2-induced cell injury. Hence, CsA may be developed as a candidate drug to prevent or treat myocardial ischemia reperfusion injury.
    Keywords:  Cardiomyocytes; CsA; H2O2; NF-κB; Oxidative stress
    DOI:  https://doi.org/10.1007/s13239-019-00404-7
  9. Int J Mol Sci. 2019 Feb 02. pii: E650. [Epub ahead of print]20(3):
    Jaworski S, Strojny B, Sawosz E, Wierzbicki M, Grodzik M, Kutwin M, Daniluk K, Chwalibog A.
      Due to the development of nanotechnologies, graphene and graphene-based nanomaterials have attracted immense scientific interest owing to their extraordinary properties. Graphene can be used in many fields, including biomedicine. To date, little is known about the impact graphene may have on human health in the case of intentional exposure. The present study was carried out on U87 glioma cells and non-cancer HS-5 cell lines as in vitro model and U87 tumors cultured on chicken embryo chorioallantoic membrane as in vivo model, on which the effects of pristine graphene platelets (GPs) were evaluated. The investigation consisted of structural analysis of GPs using transmission electron microscopy, Fourier transmission infrared measurements, zeta potential measurements, evaluation of cell morphology, assessment of cell viability, investigation of reactive oxygen species production, and investigation of mitochondrial membrane potential. The toxicity of U87 glioma tumors was evaluated by calculating the weight and volume of tumors and performing analyses of the ultrastructure, histology, and protein expression. The in vitro results indicate that GPs have dose-dependent cytotoxicity via ROS overproduction and depletion of the mitochondrial membrane potential. The mass and volume of tumors were reduced in vivo after injection of GPs. Additionally, the level of apoptotic and necrotic markers increased in GPs-treated tumors.
    Keywords:  apoptosis; mitochondria; oxidative stress; pristine graphene
    DOI:  https://doi.org/10.3390/ijms20030650
  10. Mol Med Rep. 2019 Jan 30.
    Qin QJ, Cui LQ, Li P, Wang YB, Zhang XZ, Guo ML.
      Rhynchophylline (RP), the primary active ingredient of Uncaria rhynchophylla, has an anti‑hypertensive effect and protects against ischemia‑induced neuronal damage. The present study aimed to examine the roles and mechanisms of RP in myocardial ischemia‑reperfusion (MI/R) injury of rat cardiomyocytes. Cell viability, reactive oxygen species, mitochondrial membrane potential (MMP) and cell apoptosis were examined by a Cell Counting Kit‑8 assay and flow cytometry, respectively. An ELISA was performed to assess the expression of oxidative stress markers. Spectrophotometry was used to detect the degree of mitochondrial permeability transition pore (mPTP) openness. Western blotting and reverse transcription‑ quantitative polymerase chain reaction assays were used to evaluate the associated protein and mRNA expression, respectively. The present results demonstrated that RP increased the cell viability of MI/R‑induced cardiomyocytes, and suppressed the MI/R‑induced apoptosis of cardiomyocytes. Additionally, RP modulated the Ca2+ and MMP levels in MI/R‑induced cardiomyocytes. Furthermore, RP decreased the oxidative stress and mPTP level of MI/R‑induced cardiomyocytes. It was additionally observed that RP affected the apoptosis‑associated protein expression and regulated the mitochondrial‑associated gene expression in MI/R‑induced cardiomyocytes. In conclusion, RP ameliorated MI/R injury through the modulation of mitochondrial mechanisms. The potential effects of RP on the protection of MI/R‑induced apoptosis of cardiomyocytes suggest that RP may be an effective target for MI/R therapy.
    DOI:  https://doi.org/10.3892/mmr.2019.9908
  11. BMC Ophthalmol. 2019 Feb 04. 19(1): 42
    Qiu X, Rong X, Yang J, Lu Y.
      BACKGROUND: To compare the protective effects of the histone deacetylase inhibitors (HDACis) β-hydroxybutyrate (βOHB), trichostatin A (TSA), suberoylanilide hydroxamic acid (SAHA) and valproic acid (VPA) on human lens epithelial cells(HLECs) following ultraviolet-B (UVB) exposure.METHODS: HLECs were divided into subgroups: four HDACi groups, a control group, a UVB-treated group and a DMSO group (cells treated with DMSO and UVB irradiation). In the HDACi groups, HLECs were cultured with different concentrations of HDACis 12 h prior to UVB irradiation. The protective effects of the HDACis were evaluated by assessing apoptosis rates, cell activity and expression levels of genes associated with apotosis (caspase-3, Bcl-2, BAX, SOD1, FOXO3A and MT2). The levels of superoxide dismutase (SOD), reactive oxygen species (ROS), malondialdehyde (MDA) and total antioxidant capacity (T-AOC) were detected in order to evaluate oxidative stress.
    RESULTS: The results showed that SAHA (1 μmol/L, 2 μmol/L) and TSA (0.2 μmol/L) had mild protective effects on cell viability. βOHB (4 mmol/L) and TSA (0.2 mol/L) demonstrated protective effects on BCL-2 expression. TSA (0.2 mol/L) showed protective effects on SOD1 expression. TSA (0.2 mol/L) and SAHA (1 μmol/L) suppressed BAX and caspase-3 expression. TSA (0.2 mol/L, 0.8 mol/L) and SAHA (1 μmol/L, 2 μmol/L) suppressed the expression of FOXO3A and MT2. SOD levels were increased after treatment with βOHB (4 mmol/L), SAHA (8 μmol/L) and TSA (0.1 mol/L, 0.2 mol/L). T-AOC levels were increased in UVB-treated HLECs after treatment with SAHA (2 μmol/L). MDA levels decreased in UVB-treated HLECs following treatment with TSA (0.2 mol/L, 0.8 mol/L). ROS levels decreased in UVB-treated HLECs following treatment with βOHB (4 mmol/L), SAHA (1 μmol/L, 2 μmol/L) and TSA (0.2 mol/L). Western blotting results demonstrated that SOD1 levels significantly increased in the βOHB (4 mmol/L), SAHA (1 μmol/L, 2 μmol/L), TSA (0.1 mol/L, 0.2 mol/L) and VPA (5 mmol/L) groups. Only SAHA (1 μmol/L) had an anti-apoptotic effect on UVB-treated HLECs.
    CONCLUSIONS: Our findings indicate that low concentrations of HDACis (1 μmol/L of SAHA) mildly inhibit oxidative stress, thus protecting HLECs from oxidation. These results may suggest that there is a possibility to explore the clinical applications of HDACis for treatment and prevention of cataracts.
    Keywords:  Beta hydroxybutyrate; Histone deacetylase inhibitors; Lens epithelial cells; Oxidative damage; Suberoylanilide hydroxamic acid; Trichostatin a; Ultraviolet-B exposure; Valproic acid
    DOI:  https://doi.org/10.1186/s12886-019-1056-7
  12. Eur J Pharmacol. 2019 Feb 01. pii: S0014-2999(19)30043-3. [Epub ahead of print]
    Guo S, Zhang YY, Peng JJ, Li YQ, Liu WN, Tang MX, Zhang XJ, Yang J, Peng J, Luo XJ.
      Methyl protodioscin (MPD) is reported to relieve angina pectoris and myocardial ischemia, and mitochondrial E3 ubiquitin ligase 1 (Mul1) plays a key role in maintaining mitochondrial functions. Bioinformatic analysis shows potential interactions between MPD and Mul1. This study aims to explore whether MPD could protect rat brain against ischemia/reperfusion (I/R) injury through regulation of Mul1/ superoxide dismutase 2 (SOD2) pathway. The SD rat brains were subjected to 2h of ischemia following by 24h of reperfusion, which showed I/R injury (increase in neurological deficit score and infarct volume), up-regulation of Mul1 and down regulation of SOD2, these phenomena were attenuated by MPD treatment (3 or 10mg/kg, i.g.). Consistently, in cultured HT22 cells, hypoxia-reoxygenation (H/R) treatment induced cellular injury (apoptosis and LDH release) concomitant with up-regulation of Mul1 and down regulation of SOD2, these phenomena were blocked in the presence of MPD (5μM). Knockdown of Mul1 could also decrease SOD2 protein levels in HT22 cells accompanied by alleviation of H/R injury (reduction of apoptosis and LDH release). In agreement with the change of SOD2, reactive oxygen species generation was increased in H/R-treated HT22 cells while decreased in the presence of MPD. Based on these observations, we conclude that upregulation of Mul1 in rat brain contributes to cerebral I/R injury via suppression of SOD2 and that MPD protects rat brain from I/R injury through a mechanism involving regulation of Mul1/SOD2 pathway.
    Keywords:  brain injury; ischemia/reperfusion; methyl protodioscin; mitochondrial E3 ubiquitin ligase 1; superoxide dismutase 2
    DOI:  https://doi.org/10.1016/j.ejphar.2019.01.057
  13. J Enzyme Inhib Med Chem. 2019 Dec;34(1): 394-404
    Wang X, Tang D, Zou Y, Wu X, Chen Y, Li H, Chen S, Shi Y, Niu H.
      Mitochondria play important roles in the development of diabetic kidney disease (DKD). The SS peptide is a tetrapeptide that is located and accumulated in the inner mitochondrial membrane; it reduces reactive oxygen species (ROS) and prevents mitochondrial dysfunction. Podocytes are key cellular components in DKD progression. However, whether the SS peptide can exert renal protection through podocytes and the mechanism involved are unknown. In the present study, we explored the mechanisms of the SS peptide on podocyte injury in vivo and in vitro. Compared with the control group, the glomerular podocyte number and expression of WT1 were significantly reduced and TUNEL-positive podocytes were significantly increased in renal tissues in the diabetic rat. These effects were further exacerbated by hypochlorite-modified albumin (HOCl-alb) challenge but prevented by SS-31. In vitro, SS-31 blocked apoptosis in podocyte cell line induced by HOCl-alb. SS-31 prevented oxidative stress and mitochondria-dependent apoptosis signalling by HOCl-alb in vivo and in vitro, as evidenced by the release of cytochrome c (cyt c), binding of apoptosis activated factor-1 (Apaf-1) and caspase-9, and activation of caspases. These data suggest that SS-31 may prevent podocyte apoptosis, exerting renal protection in diabetes mellitus, probably through an apoptosis-related signalling pathway involving oxidative stress and culminating in mitochondria.
    Keywords:  Podocyte apoptosis; antioxidant peptide; diabetic kidney disease; hypochlorite-modified albumin; mitochondria
    DOI:  https://doi.org/10.1080/14756366.2018.1488697
  14. Cell Death Dis. 2019 Feb 04. 10(2): 96
    Li L, Su Z, Zou Z, Tan H, Cai D, Su L, Gu Z.
      Heat stroke has increased in frequency worldwide in recent years and continues to have a high morbidity and mortality. Identification of the mechanisms mediating heat stoke is important and necessary. Our preliminary study revealed heat stress (HS)-induced apoptosis of vascular endothelial cells was associated with reactive oxygen species (ROS)-induced p53 translocation into mitochondria. Previous studies have suggested the prolyl-isomerase Pin1 regulates p53 functioning through specific binding to p53 phosphorylation sites. Based on these studies, we presumed Pin1 is a key intermediate in regulation of mitochondrial p53 translocation through a HS-induced ROS-p53 transcription-independent apoptosis pathway. In this context, we revealed p53 had a crucial role in a HS-induced mitochondrial apoptotic pathway, where p53 protein rapidly translocated into mitochondria in endothelial cells both in vitro and in vivo. In particular, HS caused an increase in p53 phosphorylation at Ser46 that facilitated interactions with phosphorylation-dependent prolyl-isomerase Pin1, which has a key role in promoting HS-induced localization of p53 to mitochondria. Furthermore, we also found ROS production was a critical mediator in HS-induced Pin1/p53 signaling and was involved in regulating mitochondrial apoptosis pathway activation. Therefore, we have contributed to our profound understanding of the mechanism underlying HS-induced endothelial dysfunction in an effort to reduce the mortality and morbidity of heat stroke.
    DOI:  https://doi.org/10.1038/s41419-019-1316-8
  15. Int J Mol Sci. 2019 Feb 01. pii: E630. [Epub ahead of print]20(3):
    Yang SH, Li P, Yu LH, Li L, Long M, Liu MD, He JB.
      Cadmium (Cd) is harmful for humans and animals, especially for the reproductive system. However, the mechanism of its toxicity has not been elucidated, and how to alleviate its toxicity is very important. This study aimed to explore the role and mechanism of action of sulforaphane (SFN) in protecting mouse Leydigs (TM3) cells from cadmium (Cd)-induced damage. The half-maximal inhibitory concentration (IC50) of Cd and the safe doses of SFN were determined using a methyl thiazolyl tetrazolium (MTT) assay. The testosterone secretion from TM3 cells was measured using the enzyme-linked immunosorbent assay. The intracellular oxidative stress was evaluated using corresponding kits. The cell apoptosis was detected using flow cytometry. The mRNA expression of genes associated with NF-E2-related factor 2 (Nrf2)/antioxidant response element (ARE) signaling was detected using reverse transcription⁻polymerase chain reaction, including Nrf2, heme oxygenase I (HO-1), glutathione peroxidase (GSH-Px), NAD(P)H:quinone acceptor oxidoreductase 1 (NQO1), and γ-glutamylcysteine synthetase (γ-GCS). The protein expression of Nrf2, GSH-Px, HO-1, γ-GCS, and NQO1 was detected using Western blot analysis. The results showed that the IC50 of Cd to TM3 cells was 51.4 µmol/L. SFN reduced the release of lactate dehydrogenase from Cd-exposed cells. Cd + SFN 2.5 treatment significantly elevated testosterone concentration compared with the Cd group (p < 0.05). SFN significantly increased total superoxide dismutase (T-SOD) and GSH-Px activity and GSH content in Cd-treated cells (p < 0.05; p < 0.01), inhibited the production of malondialdehyde or reactive oxygen species caused by Cd (p < 0.05; p < 0.01), and reduced the apoptotic rate of Cd-induced TM3 cells (p < 0.01). SFN upregulated the mRNA expression of Nrf2, GSH-Px, HO-1, NQO1, and γ-GCS in Cd-treated cells, indicating the protective effect of SFN against Cd-induced oxidative stress or cell apoptosis by activating the Nrf2/ARE signaling pathway.
    Keywords:  Nrf2/ARE signaling pathway; SFN; TM3 cell; cadmium; oxidative damage
    DOI:  https://doi.org/10.3390/ijms20030630
  16. Free Radic Biol Med. 2019 Feb 05. pii: S0891-5849(18)32288-3. [Epub ahead of print]
    Rahman A, Pallichankandy S, Thayyullathil F, Galadari S.
      Natural products are a major source of potential anticancer agents, and in order to develop improved and more effective cancer treatments, there is an immense need in exploring and elucidating their mechanism of action. Sanguinarine (SNG), a quaternary benzophenanthridine alkaloid, has been shown to induce cytotoxicity in various human cancers and suppresses various pro-tumorigenic processes such as invasion, angiogenesis, and metastasis in different cancers. Lack of understanding the anticancer mechanism(s) of SNG has impeded the development of this molecule as a potential anticancer agent. Earlier, we have reported that SNG induces reactive oxygen species (ROS)-dependent ceramide (Cer) generation and Akt dephosphorylation, leading to the induction of apoptosis in human leukemic cells. In the present study, we demonstrate that SNG has potent anti-proliferative activity against prostate cancer cells. Our data suggest that SNG induces Cer generation via inhibiting acid ceramidase and glucosylceramide synthase, two important enzymes involved in Cer metabolism. Furthermore, we demonstrate that SNG induces ROS-depended extracellular signal-regulated kinase1/2 (ERK1/2) phosphorylation, and prostate apoptosis response-4 (Par-4) cleavage, leading to the induction of apoptosis in human prostate cancer cells. Overall, our findings provide molecular insight into the role of ROS signaling in the anticancer mechanism(s) of SNG. This may provide the basis for its use as a nontoxic and an effective therapeutic agent in the treatment of prostate cancer.
    Keywords:  Apoptosis; Ceramide; Extracellular signal-regulated kinase1/2; Hydrogen peroxide; Prostate apoptosis response-4; Sanguinarine
    DOI:  https://doi.org/10.1016/j.freeradbiomed.2019.01.039
  17. PeerJ. 2019 ;7 e6383
    Tang Z, Yang C, Zuo B, Zhang Y, Wu G, Wang Y, Wang Z.
      Background: Taxifolin (TAX), is an active flavonoid, that plays an underlying protective role on the cardiovascular system. This study aimed to evaluate its effect and potential mechanisms on myocardial ischemia/reperfusion (I/R) injury.Methods: Healthy rat heart was subjected to I/R using the Langendorff apparatus. Hemodynamic parameters, including heart rate, left ventricular developed pressure (LVDP), maximum/minimum rate of the left ventricular pressure rise (+dp/dt max and -dp/dt min) and rate pressure product (RPP) were recorded during the perfusion. Histopathological examination of left ventricular was measured by hematoxylin-eosin (H&E) staining. Creatine kinase-MB (CK-MB) and lactate dehydrogenase (LDH) activities in the effluent perfusion, and the levels of malondialdehyde (MDA), superoxide dismutase (SOD), and glutathione peroxidase (GSH-PX) in the tissue were assayed. Apoptosis related proteins, such as B-cell lymphoma-2 (Bcl-2), Bcl2-associated X (Bax), and cytochrome c (Cyt-c) were also assayed by ELISA. Western blot was employed to determine apoptosis-executive proteins, including caspase 3 and 9. Transferase-mediated dUTP-X nick end labeling assay was performed to evaluate the effect TAX on myocardial apoptosis.
    Results: Taxifolin significantly improved the ventricular functional recovery, as evident by the increase in LVDP, +dp/dt max, -dp/dt min and RPP, the levels of SOD, GSH-PX were also increased, but those of LDH, CK-MB, and MDA were decreased. Furthermore, TAX up-regulated the Bcl-2 protein level but down-regulated the levels of Bax, Cyt-c, caspase 3 and 9 protein, thereby inhibits the myocardial apoptosis.
    Discussion: Taxifolin treatment remarkably improved the cardiac function, regulated oxidative stress and attenuated apoptosis. Hence, TAX has a cardioprotective effect against I/R injury by modulating mitochondrial apoptosis pathway.
    Keywords:  Apoptosis; I/R; Langendorff; Oxidative stress; Taxifolin
    DOI:  https://doi.org/10.7717/peerj.6383
  18. Eur J Pharmacol. 2019 Feb 01. pii: S0014-2999(19)30086-X. [Epub ahead of print]
    Thu PM, Zheng ZG, Zhou YP, Wang YY, Zhang X, Jing D, Cheng HM, Li J, Li P, Xu X.
      Despite the massive efforts to develop the treatment of pancreatic cancers, no effective application exhibits satisfactory clinical outcome. Macropinocytosis plays a critical role for continuous proliferation of pancreatic ductal adenocarcinoma (PDAC). In this study, we generated a screening method and identified phellodendrine chloride (PC) as a potential macropinocytosis inhibitor. PC significantly inhibited the viability of KRAS mutant pancreatic cancer cells (PANC-1 and MiaPaCa-2) in a dose-dependent manner; however, it did not affect the wild type KRAS pancreatic cancer cells (BxPC-3). Further experiments indicated that PC reduced the growth of PANC-1 cells through inhibition of macropinocytosis and diminishing the intracellular glutamine level. Disruption of glutamine metabolism led to enhance the reactive oxygen species level and induce mitochondrial membrane potential depolarization in PANC-1 cells. PC treatment caused increased Bax and decreased Bcl-2 expression, along with the activation of cleaved caspase-3, 7, 9 and cleaved-PARP, thus induced mitochondrial apoptosis. Moreover, PC inhibited macropinocytosis in vivo and effectively reduced the growth of PANC-1 xenograft tumors. All together, we demonstrated that inhibition of macropinocytosis might be an effective strategy to treat pancreatic cancers. Thus, PC could be a potential compound with improved therapeutic efficacy in patients with pancreatic cancers.
    Keywords:  KRAS mutated pancreatic cancers; Phellodendrine chloride; glutamine metabolism; macropinocytosis; mitochondrial apoptosis
    DOI:  https://doi.org/10.1016/j.ejphar.2019.01.060
  19. Eur J Sport Sci. 2019 Feb 08. 1-10
    Farzanegi P, Dana A, Ebrahimpoor Z, Asadi M, Azarbayjani MA.
      Non-alcoholic fatty liver disease (NAFLD) is a common chronic liver disorder which is associated with accumulation of fats in the liver. It causes a wide variety of pathological effects such as non-alcoholic steatohepatitis (NASH) and cirrhosis, insulin resistance, obesity, hypertension, dyslipidaemia, diabetes and cardiovascular disease. The molecular mechanisms that cause the initiation and progression of NAFLD are not fully understood. Oxidative stress (OS) induced by reactive oxygen species (ROS) and inflammation are likely a significant mechanism which can lead to hepatic cell death and tissue injury. Mitochondrial abnormalities, down-regulation of several antioxidant enzymes, glutathione (GSH) depletion and decreased activity of GSH-dependent antioxidants, accumulation of leukocytes and hepatic inflammation are the major sources of ROS overproduction in NAFLD. Excessive production of ROS suppresses the capacity of other antioxidant defence systems in NAFLD and causes further oxidative damage. Regular exercise can be considered as an effective strategy for treatment of NAFLD. It improves NAFLD by reducing intrahepatic fat content, increasing β-oxidation of fatty acids, inducing hepato-protective autophagy, overexpressing peroxisome proliferator-activated receptor- γ (PPAR-γ), as well as attenuating hepatocyte apoptosis and increasing insulin sensitivity. Exercise training also suppresses ROS overproduction and OS in NAFLD via up-regulation of several antioxidant enzymes and anti-inflammatory mediators. Therefore, an understanding of these molecules and signalling pathways gives us valuable information about NAFLD progression and a method for developing a suitable clinical treatment. This review aimed to evaluate sources of ROS and OS in NAFLD and the molecular mechanisms involved in the beneficial effects of exercises on NAFLD.
    Keywords:  Exercise; NAFLD; inflammation; oxidative stress; reactive oxygen species
    DOI:  https://doi.org/10.1080/17461391.2019.1571114
  20. Food Chem Toxicol. 2019 Feb 05. pii: S0278-6915(19)30060-2. [Epub ahead of print]
    Thilakarathna WPDW, Rupasinghe HPV.
      Seven selected microbial metabolites of proanthocyanidins (MMP), 3-phenylpropionic, 4-hydroxyphenyl acetic, 3-(4-hydroxyphenyl) propionic, ρ-coumaric, benzoic acid, pyrogallol (PG), and pyrocatechol (PC) were evaluated for their ability to reduce chemical carcinogen-induced toxicity in human lung epithelial cells (BEAS-2B) and human fetal hepatic cells (WRL-68). Cells pre-treated with MMP were exposed to a known chemical carcinogen, 4-[(acetoxymethyl) nitrosamino]-1-(3-pyridyl)-1-butanone (NNKOAc) to assess MMP-mediated cytoprotection and reduction of DNA damage. PG in BEAS-2B and PC in WRL-68 cells mitigated the NNKOAc-induced cytotoxicity. Pre-incubation of PG depicted significant protection against NNKOAc-induced DNA damage in BEAS-2B cells. PC in WRL-68 cells showed similar activity. To understand the mechanisms of PG- and PC-mediated DNA damage reduction, the effect on DNA damage response (DDR) proteins, cellular reactive oxygen species (ROS), total antioxidant capacity (TAC), glutathione peroxidase (GPx), and caspase activity were studied. PG and PC alter the DDR and may promote ATR-Chk1 and ATM-Chk2 pathways, respectively. Cellular oxidative stress induced by NNKOAc was mitigated by PG and PC through enhanced GPx expression and TAC. PG and PC suppressed the activation of the extrinsic apoptotic pathway (caspase 3 and 8) provoked by NNKOAc. MMP are beneficial in chemoprevention by reducing cellular DNA damage.
    Keywords:  Antioxidant; Cancer; Carcinogen; DNA damage; Microbial metabolites; Proanthocyanidin
    DOI:  https://doi.org/10.1016/j.fct.2019.02.010
  21. Oxid Med Cell Longev. 2019 ;2019 6595189
    Liu F, Huang X, Luo Z, He J, Haider F, Song C, Peng L, Chen T, Wu B.
      In order to use stem cells as a resource for tissue regeneration, it is necessary to induce expansion in vitro. However, during culture, stem cells often lose functional properties and become senescent. Increasing evidence indicates that hypoxic preconditioning with physiological oxygen concentration can maintain the functional properties of stem cells in vitro. The purpose of the current study was to test the hypothesis that hypoxic preconditioning with physiological oxygen concentration can maintain the functional properties of stem cells in culture by reducing oxidative stress. In vitro studies were performed in primary human dental pulp cells (hDPCs). Reduced levels of oxidative stress and increased cellular "stemness" in response to physiological hypoxia were dependent upon the expression of reactive oxygen species (ROS). Subsequently, RNA-sequencing analysis revealed the increased expression of phosphoinositide 3-kinase (PI3K)/Akt signaling in culture, a pathway which regulates oxidative stress. Furthermore, we found evidence that PI3K/Akt signaling might affect intracellular ROS production by negatively regulating expression of the downstream protein Forkhead Box Protein O1 (FOXO1) and Caspase 3. Collectively, our data show that the PI3K/Akt pathway is activated in response to hypoxia and inhibits oxidative stress in a ROS-dependent manner. This study identified redox-mediated hypoxic preconditioning regulatory mechanisms that may be significant for tissue regeneration.
    DOI:  https://doi.org/10.1155/2019/6595189
  22. Toxicol Res (Camb). 2019 Jan 01. 8(1): 15-24
    Xiao Y, Zeng M, Yin L, Li N, Xiao F.
      Previous evidence revealed significant elevated liver cancer mortality in the areas where water was contaminated with hexavalent chromium [Cr(vi)], which highlighted that we should pay more attention to Cr(vi)-induced cytotoxicity in hepatocytes. We found that Clusterin (CLU) was up-regulated in Cr(vi)-exposed L-02 hepatocytes, but the role CLU played in Cr(vi)-induced cytotoxicity has never been explored. In the present study, we demonstrate Cr(vi) targeted mitochondrial respiratory chain complex I (MRCC I) activity and induced reactive oxygen species (ROS) accumulation, which caused mitochondrial damage that was characterized by the increase of permeability transition pore (PTP) open rate, the collapse of mitochondrial membrane potential (MMP), and the release of apoptosis-inducing factor (AIF) and Cytochrome C (Cyt C) from mitochondria to cytoplasm, which then induced cell viability loss and increased aspartate transaminase (AST)/alanine transaminase (ALT) leakage. We reveal that Cr(vi) may regulate CLU expression through the ROS-ataxia telangiectasia mutant (ATM)-insulin-like growth factor 1 (IGF-1) axis, and CLU expression was positively correlated to MRCC I activity. We further confirmed that CLU may regulate MRCC I activity via modulating its subunit nicotinamide adenine dinucleotide dehydrogenase (ubiquinone) Fe-S protein 3 (NDUFS3) expression. By the establishment of CLU over-expression cells, we found that over-expression of CLU alleviated Cr(vi)-induced MRCC I inhibition and further rescued cell viability loss and reduced AST and ALT leakage. Thus, we reached the conclusion that the CLU-induced increase of MRCC I activity protected against Cr(vi)-induced cytotoxicity. The present research will provide new experimental evidence for thoroughly clarifying the cytotoxicity and the carcinogenic mechanism of Cr(vi).
    DOI:  https://doi.org/10.1039/c8tx00231b
  23. Mol Med Rep. 2019 Jan 28.
    Li S, Sun C, Gu Y, Gao X, Zhao Y, Yuan Y, Zhang F, Hu P, Liang W, Cao K, Zhang J, Wang Z, Ye J.
      Increasing evidence has indicated that mutations of isocitrate dehydrogenase 1/2 (IDH1/2) contribute to the metabolic reprogramming of cancer cells; however their functions in lipid metabolism remain unknown. In the present study, the parental and IDH1 (R132H/+) mutant HCT116 cells were treated with various concentrations of oleic acid (OA) or palmitic acid (PA) in the presence or absence of glucose. The results demonstrated that mutation of IDH1 exacerbated the effects of OA and PA on cell viability and apoptosis, and consistently elevated the production of reactive oxygen species in HCT116 cells, particularly in the absence of glucose. Furthermore, mutation of IDH1 inhibited the rate of fatty acid oxidation (FAO), but elevated the glucose consumption in HCT116 cells. The results of immunoblotting and reverse transcription‑quantitative polymerase chain reaction (RT‑qPCR) indicated that the expression of glucose transporter 1 was upregulated, whereas that of carnitine palmitoyl transferase 1 was downregulated in IDH1 mutant HCT116 cells. Although mitochondrial DNA quantification demonstrated that mutation of IDH1 had no effect on the quantity of mitochondria, immunoblotting and RT‑qPCR revealed that mutation of IDH1 in HCT116 cells significantly downregulated the expression of cytochrome c (CYCS) and CYCS oxidase IV, two important components in mitochondrial respiratory chain. These results indicated that mutation of IDH1 aggravated the fatty acid‑induced oxidative stress in HCT116 cells, by suppressing FAO and disrupting the mitochondrial respiratory chain. The results of the present study may provide novel insight into therapeutic strategies for the treatment of cancer types with IDH mutation.
    DOI:  https://doi.org/10.3892/mmr.2019.9903
  24. Food Chem Toxicol. 2019 Feb 05. pii: S0278-6915(19)30018-3. [Epub ahead of print]
    Sahu R, Dua TK, Das S, De Feo V, Dewanjee S.
      The present investigation has been undertaken to reveal the protective mechanism of polyphenolics extract of whole wheat grains (WWGPE), ferulic acid and apigenin against doxorubicin (Dox)-induced cardio-toxicity. WWGPE, apigenin, and ferulic acid exhibited concentration dependent cyto-protective effect against Dox (1 μM) in rat cardiomyocytes. Dox treatment significantly (p < 0.01) induced oxidative stress in the myocardial cells via excessive ROS production, increase in iNOS expression, NADPH oxidase activation, Nrf-2/HO-1 impairment, and inactivation of cellular redox defense system. In addition, Dox significantly (p < 0.01) activated MAP kinases, NF-κB, and apoptosis in cardiac cells; while, significant (p < 0.01) impairment in PI3K/Akt/mTOR signaling was observed in Dox-treated myocardial cells. On the other hand, WWGPE, apigenin, and ferulic acid significantly (p < 0.05-0.01) attenuated Dox-induced redox stress and oxidative stress-mediated signal transduction in myocardial cells. WWGPE, apigenin, and ferulic acid treatment also could significantly (p < 0.05-0.01) reinstate Dox-mediated changes in blood parameters in rats. Histological assessments were in agreement with the biochemical findings. Results showed that, WWGPE exhibited better cardio-protective effect over ferulic acid and apigenin, which may be due to the synergy between the comprising compounds and better oral bioavailability of dietary antioxidant molecules from whole phenolic extract.
    Keywords:  Apoptosis; Cardiomyopathy; Doxorubicin; Oxidative stress; Wheat phenolics; Whole wheat grains
    DOI:  https://doi.org/10.1016/j.fct.2019.01.034
  25. Toxicology. 2019 Jan 31. pii: S0300-483X(18)30225-7. [Epub ahead of print]
    Rebleanu D, Gaidau C, Voicu G, Constantinescu CA, Mansilla Sánchez C, Rojas TC, Carvalho S, Calin M.
      The potential human health risks following the exposure to inorganic nanoparticles (NPs) is a very important issue for their application in leather finishing industry. The aim of our study was to investigate the cytotoxic effect of silver (Ag)/titanium dioxide (TiO2) NPs on human cells. Photocatalytic NPs were prepared by electrochemical deposition of Ag on the surface of TiO2 and nitrogen (N)-TiO2 NPs and, subsequently, physico-chemical characterized. Then, a set of experiments have been performed to study the cytotoxicity and cell death mechanisms involved, the changes in cell morphology and the production of ROS induced in human keratinocytes (HaCaT) and human lung epithelial cells (A549) by exposure to NPs. Moreover, the changes in major signaling pathways and the inflammatory response induced by Ag/N-TiO2 NPs in A549 cells were investigated. The data showed that cell death by late apoptosis/necrosis is induced in cells as function of the dose and the type of NPs and is characterized by morphological changes and cytoskeletal disorganization and an increase in reactive oxygen species (ROS) production. The exposure of A549 cells to Ag/N-TiO2 NPs determine the activation of ERK1/2 MAP-kinase pathway and the release of pro-inflammatory mediators CXCL1, GM-CSF and MIF, known to be involved in the recruitment of circulating neutrophils and monocytes. In conclusion, due to its cyto-friendly properties as compared to Ag/TiO2 NPs, we consider Ag/N-TiO2 NPs suitable for leather surface finishing with some caution regarding the limit of dose handled, that has to be below 0.5 mg/ml.
    Keywords:  Ag/N-TiO(2); Ag/TiO(2); cytokines; cytotoxicity; intracellular signalling; nanoparticles
    DOI:  https://doi.org/10.1016/j.tox.2019.01.013
  26. Neurochem Res. 2019 Feb 06.
    Tabeshpour J, Mehri S, Abnous K, Hosseinzadeh H.
      Acrylamide (ACR) is extensively used in industrial areas and has been demonstrated to induce neurotoxicity via oxidative stress and apoptosis. In this study, we assessed the probable protective effects of thymoquinone (TQ), an active constituent of Nigella sativa, against ACR-induced neurotoxicity. ACR (50 mg/kg, i.p., for 11 days) and TQ (2.5, 5 and 10 mg/kg, i.p., for 11 days) were administered to rats. On 12th day, gait score was examined and rats were sacrificed. Malondialdehyde (MDA) and reduced glutathione (GSH) contents were determined in sciatic nerve. Furthermore, western blotting was conducted. The exposure of rats to ACR caused severe gait disabilities. The MDA and GSH contents were increased and decreased, respectively. ACR decreased P-ERK/ERK ratio and myelin basic protein (MBP) content, but significantly increased P-JNK/JNK, P-P38/P38, Bax/Bcl-2 ratios and caspase 3 and 9 levels. Concurrently administration of TQ (5 and 10 mg/kg) with ACR, prevented gait abnormalities and meaningfully reduced MDA and elevated the GSH contents. Furthermore, TQ (5 mg/kg) elevated the P-ERK/ERK ratio and MBP content while reduced the P-JNK/JNK, P-P38/P38 ratios and apoptotic markers. MAP kinase and apoptosis signaling pathways were involved in ACR-induced neurotoxicity in rat sciatic nerve and TQ significantly reduced ACR neurotoxicity. TQ afforded neuroprotection, in part, due to its anti-oxidative stress and anti-apoptotic mechanisms.
    Keywords:  Acrylamide; Apoptosis; MAP kinase signaling pathway; Neuroprotective; Sciatic nerve; Thymoquinone
    DOI:  https://doi.org/10.1007/s11064-019-02741-4
  27. J Nanobiotechnology. 2019 Feb 05. 17(1): 24
    Lee K, Lee J, Kwak M, Cho YL, Hwang B, Cho MJ, Lee NG, Park J, Lee SH, Park JG, Kim YG, Kim JS, Han TS, Cho HS, Park YJ, Lee SJ, Lee HG, Kim WK, Jeung IC, Song NW, Bae KH, Min JK.
      BACKGROUND: Silica nanoparticles (SiNPs) are widely used for biosensing and diagnostics, and for the targeted delivery of therapeutic agents. Safety concerns about the biomedical and clinical applications of SiNPs have been raised, necessitating analysis of the effects of their intrinsic properties, such as sizes, shapes, and surface physicochemical characteristics, on human health to minimize risk in biomedical applications. In particular, SiNP size-associated toxicological effects, and the underlying molecular mechanisms in the vascular endothelium remain unclear. This study aimed to elucidate the detailed mechanisms underlying the cellular response to exposure to trace amounts of SiNPs and to determine applicable size criteria for biomedical application.METHODS: To clarify whether these SiNP-mediated cytotoxicity due to induction of apoptosis or necrosis, human ECs were treated with SiNPs of four different non-overlapping sizes under low serum-containing condition, stained with annexin V and propidium iodide (PI), and subjected to flow cytometric analysis (FACS). Two types of cell death mechanisms were assessed in terms of production of reactive oxygen species (ROS), endoplasmic reticulum (ER) stress induction, and autophagy activity.
    RESULTS: Spherical SiNPs had a diameter of 21.8 nm; this was further increased to 31.4, 42.9, and 56.7 nm. Hence, we investigated these effects in human endothelial cells (ECs) treated with these nanoparticles under overlap- or agglomerate-free conditions. The 20-nm SiNPs, but not SiNPs of other sizes, significantly induced apoptosis and necrosis. Surprisingly, the two types of cell death occurred independently and through different mechanisms. Apoptotic cell death resulted from ROS-mediated ER stress. Furthermore, autophagy-mediated necrotic cell death was induced through the PI3K/AKT/eNOS signaling axis. Together, the present results indicate that SiNPs within a diameter of < 20-nm pose greater risks to cells in terms of cytotoxic effects.
    CONCLUSION: These data provide novel insights into the size-dependence of the cytotoxic effects of silica nanoparticles and the underlying molecular mechanisms. The findings are expected to inform the applicable size range of SiNPs to ensure their safety in biomedical and clinical applications.
    Keywords:  Apoptosis; Autophagy; Necroptosis; ROS; Silica nanoparticles
    DOI:  https://doi.org/10.1186/s12951-019-0456-4
  28. Sci Rep. 2019 Feb 04. 9(1): 1200
    Vanneste M, Huang Q, Li M, Moose D, Zhao L, Stamnes MA, Schultz M, Wu M, Henry MD.
      Epithelial-to-mesenchymal transition (EMT) is implicated in cancer metastasis and drug resistance. Specifically targeting cancer cells in an EMT-like state may have therapeutic value. In this study, we developed a cell imaging-based high-content screening protocol to identify EMT-selective cytotoxic compounds. Among the 2,640 compounds tested, salinomycin and monensin, both monovalent cation ionophores, displayed a potent and selective cytotoxic effect against EMT-like cells. The mechanism of action of monensin was further evaluated. Monensin (10 nM) induced apoptosis, cell cycle arrest, and an increase in reactive oxygen species (ROS) production in TEM 4-18 cells. In addition, monensin rapidly induced swelling of Golgi apparatus and perturbed mitochondrial function. These are previously known effects of monensin, albeit occurring at much higher concentrations in the micromolar range. The cytotoxic effect of monensin was not blocked by inhibitors of ferroptosis. To explore the generality of our findings, we evaluated the toxicity of monensin in 24 human cancer cell lines and classified them as resistant or sensitive based on IC50 cutoff of 100 nM. Gene Set Enrichment Analysis identified EMT as the top enriched gene set in the sensitive group. Importantly, increased monensin sensitivity in EMT-like cells is associated with elevated uptake of 3H-monensin compared to resistant cells.
    DOI:  https://doi.org/10.1038/s41598-018-38019-y
  29. Anticancer Res. 2019 Feb;39(2): 687-693
    Shinada H, Watanabe T, Okudaira K, Iwase Y, Nishi K, Yumita N.
      BACKGROUND: The ultraviolet A (UVA) spectrum mainly includes the region associated with the phototoxicity of fluoroquinolone antimicrobial agents. This study investigated apoptosis induced with UVA light and enoxacin in HL-60 cells.MATERIALS AND METHODS: HL-60 cells were irradiated by UVA (1.1 mW/cm2) for 20 min in the presence or absence of enoxacin. The induction of apoptosis was investigated by analysing cell morphology, flow cytometry of annexin V-positive cells, DNA ladder formation, and caspase-3 activation.
    RESULTS: Significant induction of apoptosis, DNA fragmentation, and caspase-3 activation were observed in cells treated with both UVA and enoxacin. UVA-induced apoptosis was significantly suppressed when NaN3, a singlet oxygen scavenger, was present.
    CONCLUSION: Apoptosis was induced by the combination of UVA and enoxacin in HL-60 cells, and singlet oxygen appears to play an important role in photodynamically-induced apoptosis.
    Keywords:  Apoptosis; HL-60; caspase-3; enoxacin; reactive oxygen species; ultraviolet A
    DOI:  https://doi.org/10.21873/anticanres.13164
  30. Cells Tissues Organs. 2019 Feb 04. 1-11
    Abu Gazia M, El-Magd MA.
      This study was conducted to evaluate the potential cardioprotective effect of cardamom (CAR) against myocardial injuries induced by doxorubicin (DOX) in rats through investigation of histological alterations and the associated oxidative stress, apoptosis, inflammation, and angiogenesis. This study included 30 adult male albino rats that were randomized to 3 groups (n = 10/group): group I (control), group II (DOX) rats injected with DOX (2.5 mg/kg body weight [BW] i.p.) every other day for 2 weeks, and group III (CAR+DOX) received CAR extract (200 mg/kg BW) orally for 3 weeks, and 1 week later (starting from the 2nd week) they were injected with DOX (2.5 mg/kg BW i.p.) every other day for 2 weeks. Rats treated with DOX alone exhibited notable myocardial damage (discontinuity and disorganization of cardiac muscle fibers, mononuclear cell infiltration, and apparent increases in collagen fiber deposition) accompanied by loss of function (revealed by elevated serum levels of lactate dehydrogenase, creatine kinase, and cardiac troponin), induction of oxidative stress (indicated by higher levels of nitric oxide and malon-dialdehyde, and lower levels of superoxide dismutase, catalase, and glutathione peroxidase), apoptosis (evidenced by high caspase 3 activity and immunostaining), and inflammation (marked by high cardiac NFκB level). However, administration of CAR not only ameliorated all deleterious effects of DOX but also induced angiogenesis, as indicated by a significant increase in VEGF immunoreactivity. These data indicate that CAR could relieve DOX-induced cardiotoxicity, at least in part, via reductions in oxidative stress, apoptosis, and inflammation and increased tissue regeneration via induction of angiogenesis. Therefore, CAR could be a promising cytoprotective agent against DOX cardiotoxicity.
    Keywords:  Apoptosis; Cardamom; Cardiotoxicity; Doxorubicin; Oxidative stress
    DOI:  https://doi.org/10.1159/000496109
  31. Oxid Med Cell Longev. 2019 ;2019 4264580
    Jun JH, Shim JK, Oh JE, Shin EJ, Shin E, Kwak YL.
      Emerging evidence indicates the pronounced role of inflammasome activation linked to reactive oxygen species (ROS) in the sterile inflammatory response triggered by ischemia/reperfusion (I/R) injury. Ethyl pyruvate (EP) is an antioxidant and conveys myocardial protection against I/R injury, while the exact mechanisms remain elusive. We aimed to investigate the effect of EP on myocardial I/R injury through mechanisms related to ROS and inflammasome regulation. The rats were randomly assigned to four groups: (1) sham, (2) I/R-control (IRC), (3) EP-pretreatment + I/R, and (4) I/R + EP-posttreatment. I/R was induced by a 30 min ligation of the left anterior descending artery followed by 4 h of reperfusion. EP (50 mg/kg) was administered intraperitoneally at 1 h before ischemia (pretreatment) or upon reperfusion (posttreatment). Both pre- and post-EP treatment resulted in significant reductions in myocardial infarct size (by 34% and 31%, respectively) and neutrophil infiltration. I/R-induced myocardial expressions of NADPH oxidase-4, carnitine palmitoyltransferase 1A, and thioredoxin-interacting protein (TXNIP) were mitigated by EP. EP treatment was associated with diminished inflammasome activation (NOD-like receptor 3 (NLRP3), apoptosis-associated speck-like protein, and caspase-1) and interleukin-1β induced by I/R. I/R-induced phosphorylation of ERK and p38 were also mitigated with EP treatments. In H9c2 cells, hypoxia-induced TXNIP and NLRP3 expressions were inhibited by EP and to a lesser degree by U0126 (MEK inhibitor) and SB203580 (p38 inhibitor) as well. EP's downstream protective mechanisms in myocardial I/R injury would include mitigation of ROS-mediated NLRP3 inflammasome upregulation and its associated pathways, partly via inhibition of hypoxia-induced phosphorylation of ERK and p38.
    DOI:  https://doi.org/10.1155/2019/4264580
  32. J Inorg Biochem. 2019 Jan 25. pii: S0162-0134(18)30536-1. [Epub ahead of print]193 112-123
    Chen JC, Zhang Y, Jie XM, She J, Dongye GZ, Zhong Y, Deng YY, Wang J, Guo BY, Chen LM.
      Thioredoxin reductase (TrxR), a major component of the thioredoxin system, makes a critical role in regulating cellular redox signaling and is found to be overexpressed in many human cancer cells. TrxR has become an attractive target for anticancer agents. In this work, three Ru(II) complexes with salicylate as ligand, [Ru(phen)2(SA)] (phen = 1,10-phenanthroline, SA = salicylate, 1), [Ru(dmb)2(SA)] (dmb = 4,4'-dimethyl-2,2'-bipyridine, 2) and [Ru(bpy)2(SA)] (bpy = 2,2'-bipyridine, 3), were synthesized and characterized. The anticancer effect exerted by them was evaluated. Complex 1 was found to exhibit obvious anticancer activity, in comparison with cisplatin, against cancer cell lines, while displaying low toxicity to the normal cell line BEAS-2B. The mechanism of complex 1 cancer cell growth suppress was investigated in A549 cells. Complex 1 exerted its anticancer through inducing apoptosis and triggering cell cycle arrest at the G0/G1 phase. Complex 1 can selectively inhibit TrxR activity and thus promote the generation and accumulation of reactive oxygen species (ROS), which subsequently trigger mitochondrial dysfunction and DNA damage, activate oxidative stress-sensitive mitogen activated protein kinase (MAPK), and suppress the protein kinase B (PKB or AKT) signal pathway, resulting in apoptosis in A549 cells.
    Keywords:  Apoptosis; ROS; Ru (II) complexes; Salicylate; Thioredoxin reductase
    DOI:  https://doi.org/10.1016/j.jinorgbio.2019.01.011
  33. Sci Rep. 2019 Feb 08. 9(1): 1683
    Da X, Nishiyama Y, Tie D, Hein KZ, Yamamoto O, Morita E.
      Ou-gon, an extract from Scutellaria baicalensis Georgi root, has been shown to exhibit pronounced antifungal activity. The present study aimed to identify antifungal components of Ou-gon and to determine their mechanism of action against pathogenic fungi. Antifungal activity was assessed by the microbroth dilution method using four common human pathogenic fungi, Trichophyton rubrum, Trichophyton mentagrophytes, Aspergillus fumigatus, and Candida albicans. Components of crude Ou-gon extract were separated by reversed-phase high-performance liquid chromatography. Active antifungal components were identified by liquid chromatography-electrospray ionization tandem mass spectrometry. Terminal deoxynucleotidyl transferase dUTP nick end-labelling assay, SYTOX® green uptake assay, determination of intracellular reactive oxygen species and mitochondrial membrane potential as well as microscopy (confocal laser microscopy, scanning and transmission electron microscopy) were used to probe the mode of action. Two components with potent antifungal activity, baicalein and wogonin, were identified in Ou-gon. Baicalein showed potent antifungal activity against the four fungi tested. Wogonin displayed antifungal activity against all four fungi except C. albicans. The components are considered to induce apoptosis-like programmed cell death via hyperproduction of reactive oxygen species. This study enhances our understanding of the antifungal activity of Kampo medicine, and may contribute to the development of new and safe antifungal therapeutics.
    DOI:  https://doi.org/10.1038/s41598-019-38916-w
  34. Methods Mol Biol. 2019 ;1866 199-210
    Spallholz JE.
      Colloidal selenium, was first used to treat cancer as early as 1911 in both humans and mice. Selenium was identified as the toxic component in forage plants of sheep, cattle, and horses in the 1930s. The animal toxicity of selenium compounds was determined to be from the metabolism by animals of the elevated concentrations of Se-methylselenocysteine and selenomethionine in plants. The metabolism of both Se-methylselenocysteine and selenomethionine by animals gives rise to the metabolite, methylselenide (CH3Se-), which if in sufficient concentration oxidizes thiols and generates superoxide and other reactive oxygen species. Cancer cells that may overly express methionine gamma-lyase, or beta-lyase (methioninase), by induced viral genomic expression, are susceptible to free radical-induced apoptosis from selenomethionine or Se-methylselenocysteine supplementation.
    Keywords:  Cancer; Prevention; Selenide; Selenium; Selenomethionnie; Therapy
    DOI:  https://doi.org/10.1007/978-1-4939-8796-2_15
  35. Biol Trace Elem Res. 2019 Feb 05.
    Biswas P, Mukhopadhyay A, Kabir SN, Mukhopadhyay PK.
      Arsenic toxicity purportedly threats a broad spectrum of female reproductive functions. We investigated the remedial role of a casein- and pea protein-enriched high-protein diet (HPD) in combating the arsenic insult. Cyclic female rats maintained on standard diet (n = 6) or an isocaloric HPD (n = 6) were gavaged with As2O3 at 3 mg/kg BW/rat/day (n = 12) for 28 days. Vehicle-fed rats (n = 6) maintained on the standard diet served as the control. We monitored the estrus cycles and performed the histomorphometric analyses of the uterus and ovary. Uterine luminal epithelial (ULE) ultrastructure was appraised by scanning electron microscopy. Uterine oxidative stress was evaluated in the forms of ROS generation and activities of the ROS scavengers. The uterine apoptotic manifestation was blueprinted by Western blot analysis of caspase-3 and Bax expression. Arsenic treatment arrested the follicular maturation and disrupted the estrus cycles with a typical increase in the diestrus index. Shrunken endometrial glands and thinned microvilli density of the ULE reflected loss of cell polarity and mislaid uterine homeostasis. Increased ROS generation and attenuated activities of the ROS scavengers marked a state of uterine oxidative imbalance and loss of redox regulation. Superfluous expression of procaspase-3, cleaved caspase-3, and Bax mirrored an inflated state of uterine apoptosis. HPD supplementation, by and large, counteracted these arsenic impacts and maintained the frameworks close to the control levels. In conclusion, arsenic mediates its reproductive toxicity, at least in part, by upsetting the uterine ROS homeostasis and redox regulation. Pea proteins and casein-supplemented HPD can counteract the arsenic effects and maintain the reproductive functions.
    Keywords:  Apoptosis; Arsenic; Casein; Oxidative stress; Pea protein; Uterine microvilli
    DOI:  https://doi.org/10.1007/s12011-019-1657-2
  36. Eur J Med Chem. 2019 Jan 24. pii: S0223-5234(19)30069-8. [Epub ahead of print]166 243-255
    Güzelcan EA, Baxendale IR, Cetin-Atalay R, Baumann M.
      Two series of boehmeriasin A analogs have been synthesized in short and high yielding processes providing derivatives differing either in the alkaloid's pentacyclic scaffold or its peripheral substitution pattern. These series have enabled, for the first time, comparative studies into key biological properties revealing a new lead compound with exceptionally high activity against liver cancer cell lines in the picomolar range for both well (Huh7, Hep3B and HepG2) and poorly (Mahlavu, FOCUS and SNU475) differentiated cells. The cell death was characterized as apoptosis by cytochrome-C release, PARP protein cleavage and SubG1 cell cycle arrest. Subsequent testing associated apoptosis via oxidative stress with in situ formation of reactive oxygen species (ROS) and altered phospho-protein levels. Compound 19 decreased Akt protein phosphorylation which is crucially involved in liver cancer tumorigenesis. Given its simple synthetic accessibility and intriguing biological properties this new lead compound could address unmet challenges within liver cancer therapy.
    Keywords:  Boehmeriasin A; Cell cycle analysis; Liver cancer; Natural product analogs
    DOI:  https://doi.org/10.1016/j.ejmech.2019.01.056
  37. Mar Drugs. 2019 Feb 01. pii: E95. [Epub ahead of print]17(2):
    Zhen AX, Piao MJ, Hyun YJ, Kang KA, Madushan Fernando PDS, Cho SJ, Ahn MJ, Hyun JW.
      The skin, the largest organ in humans, is exposed to major sources of outdoor air pollution, such as fine particulate matter with a diameter ≤ 2.5 µm (PM2.5). Diphlorethohydroxycarmalol (DPHC), a marine-based compound, possesses multiple activities including antioxidant effects. In the present study, we evaluated the protective effect of DPHC on PM2.5-induced skin cell damage and elucidated the underlying mechanisms in vitro and in vivo. The results showed that DPHC blocked PM2.5-induced reactive oxygen species generation in human keratinocytes. In addition, DPHC protected cells against PM2.5-induced DNA damage, endoplasmic reticulum stress, and autophagy. HR-1 hairless mice exposed to PM2.5 showed lipid peroxidation, protein carbonylation, and increased epidermal height, which were inhibited by DPHC. Moreover, PM2.5 induced apoptosis and mitogen-activated protein kinase (MAPK) protein expression; however, these changes were attenuated by DPHC 5. MAPK inhibitors were used to elucidate the molecular mechanisms underlying these actions, and the results demonstrated that MAPK signaling pathway may play a key role in PM2.5-induced skin damage.
    Keywords:  MAPK; PM2.5; diphlorethohydroxycarmalol; human keratinocytes; skin cell damage
    DOI:  https://doi.org/10.3390/md17020095
  38. Int J Mol Sci. 2019 Feb 02. pii: E651. [Epub ahead of print]20(3):
    Park SH, Kim DS, Kim S, Lorz LR, Choi E, Lim HY, Hossain MA, Jang S, Choi YI, Park KJ, Yoon K, Kim JH, Cho JY.
      Loliolide is a monoterpenoid hydroxylactone present in freshwater algae that has anti-inflammatory and antiaging activity; however, its effects on ultraviolet-damaged skin have yet to be elucidated. This study investigated the antiapoptosis and wound-healing effects of loliolide using HaCaT cells (a human keratinocyte cell line). Loliolide inhibited the expression of reactive oxygen species (ROS) induced by ultraviolet radiation as well as wrinkle formation-related matrix metalloproteinase genes and increased the expression of the damage repair-related gene SIRT1. The apoptosis signaling pathway was confirmed by Western blot analysis, which showed that loliolide was able to reduce the expression of caspases 3, 8, and 9, which are related to ROS-induced apoptosis. In addition, Western blotting, reverse-transcription polymerase chain reaction (PCR), and real-time PCR analyses showed that loliolide enhanced the expression of the epidermal growth factor receptor signaling pathway (PI3K, AKT) and migration factors, such as K6, K16, and K17; keratinocyte growth factor; and inflammatory cytokines, such as interleukin (IL)-1, IL-17, and IL-22 expressed during the cellular scratching process, suggesting a putative wound-healing ability. Because of the antiapoptosis and antiscratching effects on skin of both loliolide and loliolide-rich Prasiola japonica ethanol extract, we consider the former to be an important compound used in the cosmeceutical industry.
    Keywords:  antiapoptosis; antioxidant activity; antiscratching effect; antiwrinkling activity
    DOI:  https://doi.org/10.3390/ijms20030651
  39. Drug Des Devel Ther. 2019 ;13 387-396
    Yao K, Zhao YF, Zu HB.
      Purpose: As a novel antidepressant drug, agomelatine has good therapeutic effect on the mood disorder and insomnia in Alzheimer's disease (AD). Recent studies have shown the neuroprotective function of agomelatine, including anti-oxidative and anti-apoptosis effect. However, it remains unclear whether agomelatine exerts neuroprotection in AD. Thus, the neuroprotective effect of agomelatine against amyloid beta 25-35 (Aβ25-35)-induced toxicity in PC12 cells was evaluated in this study.Methods: The concentration of malondialdehyde (MDA), LDH, and ROS was investigated to evaluate oxidative damage. The expression of P-tau, tau, PTEN, P-Akt, Akt, P-GSK3β, and GSK3β proteins was assessed by Western blotting. Our results demonstrated that Aβ25-35 significantly increased the content of MDA, LDH, and ROS. Meanwhile, Aβ25-35 upregulated the expression of P-tau and PTEN as well as downregulated P-Akt and P-GSK3β expression. These effects could be blocked by agomelatine pretreatment. Furthermore, luzindole, the melatonin receptor (MT) antagonist, could reverse the neuroprotective effect of agomelatine.
    Conclusion: The results demonstrated that antidepressant agomelatine might prevent the tau protein phosphorylation and oxidative damage induced by Aβ25-35 in PC12 cells by activating MT-PTEN/Akt/GSK3β signaling. This study provided a novel therapeutic target for AD in the future.
    Keywords:  Alzheimer’s disease; agomelatine; oxidative stress; tau hyperphosphorylation
    DOI:  https://doi.org/10.2147/DDDT.S182684
  40. Anticancer Res. 2019 Feb;39(2): 591-596
    Mantso T, Anestopoulos I, Lamprianidou E, Kotsianidis I, Pappa A, Panayiotidis MI.
      BACKGROUND/AIM: Several studies have documented the effects of isothiocyanates (ITCs) on cancer prevention by inducing oxidative stress, activating apoptosis, affecting cell-cycle regulation, etc. Previously, we have shown that ITCs, administered at low concentrations by the means of double-bolus are capable of potentiating cytotoxicity in human malignant melanoma (A375) cells by inducing apoptosis. The aim of the present study was to further investigate the effect of the treatment of A375 cells with ITCs on cell-cycle progression and the levels of various cell cycle regulators.MATERIALS AND METHODS: Cell-cycle analysis was performed by means of flow cytometry whereas western immunoblotting was used to determine the expression levels of these protein regulators.
    RESULTS: Our data showed an increase in the number of cells in the G2/M phase accompanied by a decrease in the G0/G1 phase, while several cell-cycle regulators were shown to be differentially expressed upon exposure to ITCs.
    CONCLUSION: ITCs induced cell-cycle arrest in A375 cells.
    Keywords:  A375 cells; Isothiocyanates; cell cycle; growth arrest; malignant melanoma
    DOI:  https://doi.org/10.21873/anticanres.13152
  41. Oxid Med Cell Longev. 2019 ;2019 1896041
    Xu L, Lin X, Guan M, Zeng Y, Liu Y.
      Diabetic neuropathy (DN) is a common and severe complication of diabetes mellitus. There is still a lack of an effective treatment to DN because of its complex pathogenesis. Thioredoxin-interacting protein (TXNIP), an endogenous inhibitor of thioredoxin, has been shown to be associated with diabetic retinopathy and nephropathy. Herein, we aim to investigate the role of TXNIP in prediabetic neuropathy and therapeutic potential of verapamil which has been shown to inhibit TXNIP expression. The effects of mediating TXNIP on prediabetic neuropathy and its exact mechanism were performed using high-fat diet- (HFD-) induced diabetic mice and palmitate-treated neurons. Our results showed that TXNIP upregulation is associated with prediabetic neuropathy in HFD-fed mice. TXNIP knockdown improved DN in HFD-induced prediabetic mice. Mechanistically, increased TXNIP in dorsal root ganglion is transferred into the cytoplasm and shuttled to the mitochondria. In cytoplasm, TXNIP binding to TRX1 results in the increased oxidative stress and inflammation. In mitochondria, TXNIP binding to TRX2 induced mitochondria dysfunction and apoptosis. TXNIP isolated from TRX2 then shuttles to the cytoplasm and binds to NLRP3, resulting in further increased TXNIP-NLRP3 complex, which induced the release of IL-1β and the development of inflammation. Thus, apoptosis and inflammation of dorsal root ganglion neuron eventually cause neural dysfunction. In addition, we also showed that verapamil, a known inhibitor of calcium channels, improved prediabetic neuropathy in the HFD-fed mice by inhibiting the upregulation of TXNIP. Our finding suggests that TXNIP might be a potential target for the treatment of neuropathy in prediabetic patients with dyslipidemia.
    DOI:  https://doi.org/10.1155/2019/1896041
  42. J Trace Elem Med Biol. 2019 Mar;pii: S0946-672X(18)30529-7. [Epub ahead of print]52 254-262
    Caglayan A, Katlan DC, Tuncer ZS, Yüce K.
      Epithelial ovarian cancer (EOC) has been associated with oxidative stress (OS) due to epithelial inflammation which makes ovaries more vulnerable to the deleterious effects of reactive oxygen species (ROS). However, antioxidant enzymes (AOEs) such as manganese-superoxide dismutase (Mn-SOD), copper,zinc-superoxide dismutase (Cu,Zn-SOD) and glutathione peroxidase (GPx1) protect cells against the biological damage of ROS-induced OS and support cancer prevention by maintaining normal cell cycle progression, inhibiting proliferation, tumor invasion, angiogenesis, inflammation or inducing apoptosis. In the present study, we aimed to measure the levels of trace elements [manganese (Mn), copper (Cu), zinc (Zn) and selenium (Se)] which are structurally and/or functionally associated with the AOEs by inductively coupled plasma/mass-spectrometry (ICP/MS) in blood samples of patients with EOC (M, n = 26) and compare the data with healthy subjects (C, n = 46). Serous EOC (M1, n = 18) data were also evaluated according to the tumor grading [well or moderately well differentiated (G 1-2) vs. poorly differentiated or undifferentiated (G3)] and staging of disease [stage I-II (SI-II) vs. stage III (SIII)]. We obtained; i) The Mn and Se levels of M were significantly lower than C, ii) only Mn levels were changed [(G3(Mn) < G 1-2 (Mn)] in M1, iii) significant correlations were observed between [Cu and Zn levels (r = 0.701, p = 0.036) in G 1-2 and (r = 0.686, p = 0.041) in G3; Cu and Se levels (r = 0.960, p = 0.000) in G3; Mn levels and Mn-SOD expression (r = 0.551, p = 0.006) in M, (r = 0.857, p = 0.007) in G 1-2 and (r = 0.690, p = 0.056) in G3; Se levels and erythrocyte GPx1 activity (r = 0.660, p = 0.053) in G 1-2 ; Se levels and erythrocyte Cu,Zn-SOD activity (r = 0.693, p = 0.038) in G3]. The study revealed that trace elements, particularly low Mn and Se levels along with high Cu/Se ratios might be of value in all histologic subtypes of EOC. Although Mn level was important in terms of discriminating tumor grades, positive correlation between Cu-Se levels was also remarkable in patients with G 1-2 tumors of M1. Moreover, high erythrocyte Cu/Se ratios might be a favourable marker for EOC.
    Keywords:  Copper; Epithelial ovarian cancer; Manganese; Selenium; Trace elements; Zinc
    DOI:  https://doi.org/10.1016/j.jtemb.2019.01.010
  43. J Card Fail. 2019 Feb 02. pii: S1071-9164(18)30313-0. [Epub ahead of print]
    Neidhardt S, Garbade J, Emrich F, Klaeske K, Borger MA, Lehmann S, Jawad K, Dieterlen MT.
      BACKGROUND: Oxidative stress due to reactive oxygen species (ROS) production is one key factor in the development of heart failure (HF). The present study investigated the thioredoxin (Trx) system, which plays a major role in antioxidant defense, in patients suffering from ischemic (ICM) or dilative cardiomyopathy (DCM).METHODS: Myocardial tissue from ICM (n=13), DCM (n=13) as well as septal tissue of patients with aortic stenosis but without diagnosed hypertrophic cardiomyopathy and subaortic stenosis (control, n=12) was analyzed for Trx1, Trx-interacting protein (TXNIP) and E3-ligase ITCH expression. Trx-reductase 1 (TXNRD1) amount and activity, cytosolic cytochrome c content and apoptosis markers were quantified by ELISA and multiplexing.
    RESULTS: Compared to controls, ITCH and Trx1 expression, TXNRD1 amount and activity were reduced and TXNIP expression was increased in ICM (ITCH: p=0.013; Trx1: p=0.028; TXNRD1 amount: p=0.035; TXNRD1 activity p=0.005; TXNIP: p=0.014) but not in DCM. A higher level of the downstream apoptosis marker caspase 9 (ICM: 582 ± 262 MFI, pto control= 0.995, DCM: 1251 ± 548 MFI, pto control=0.002, control: 561 ± 214) was detected in DCM tissue. A higher expression of Bcl-2 was found in DCM (p=0.011).
    CONCLUSION: The Trx system was impaired in ICM but not in DCM. ITCH appeared to be responsible for the downregulation of the Trx system. ROS-induced mitochondrial instability appeared to play a role in DCM.
    Keywords:  E3 ligases; Heart failure; ITCH; TXNIP; Thioredoxin; cardiomyopathy
    DOI:  https://doi.org/10.1016/j.cardfail.2019.01.017
  44. Toxicology. 2019 Feb 02. pii: S0300-483X(18)30409-8. [Epub ahead of print]
    Cherif B, Triki H, Sahnoun S, Hamden K, Sallemi A, Charfi S, Lassoued S.
      Imipenem is a β-Lactam antibiotics characterized by a larger specter activity, it's applied during hard infections in adults and children patients. Our goal is to investigate toxicity induced in male rat reproductive system following imipenem (15, 50 or 100 mg/kg) exposure compared to gentamicin (50 mg/kg) treatment. Effects of imipenem on reproductive organ weights, histoarchitecture, sperm parameters, and oxidative stress parameters were evaluated. Serum testosterone level was measured. Apoptosis and inflammatory behaviors were investigated by immunohistochemical proteins expression analysis of apoptosis regulator BAX (Bax), B-cell lymphoma 2 (Bcl-2), and interleukin-1 beta (IL-1 beta) in testis. Our results show a significant decrease in parameters related to the fertility of males such as sperm count, sperm motility, epididymis weights and serum testosterone level was observed after imipenem administration as compared to the control and gentamicin treated groups. While increased sperm abnormality was significantly observed in imipenem high doses. Oxidative stress analysis revealed a significant increase in lipid peroxidation and carbonyl groups levels in testicular tissues compared to control group. Same results were observed with superoxide dismutase and catalase activities from testicular tissues. In addition, severe testicular lesions were observed in the seminiferous tubules as well as important impairment in spermatogenesis testifying an inflammatory microenvironment confirmed by the intensive expression of IL1-beta and Bax protein by germinal cells while Bcl-2 by Leydig cells. In conclusion, imipenem treatments in particular high doses produce an oxidative stress in male reproductive organs and an inflammatory microenvironment leading to spermatogenesis dysfunction and histopathological changes in the testis.
    Keywords:  Imipenem/cilastatin; Inflammatory microenvironment; Oxidative stress; Rat Spermatogenesis
    DOI:  https://doi.org/10.1016/j.tox.2019.02.001
  45. Front Pharmacol. 2019 ;10 2
    Park H, Kang S, Nam E, Suh YH, Chang KA.
      Polygala tenuifolia Willdenow is a herb known for its therapeutic effects in insomnia, depression, disorientation, and memory impairment. In Alzheimer's disease (AD) animal model, there has been no report on the effects of memory and cognitive impairment. PSM-04, an extract from the root of P. tenuifolia Willdenow, was developed with improved bioabsorption. The present study aimed to investigate the neuroprotective effects of PSM-04 on AD and reveal the possible molecular mechanism. The neuroprotective effect of PSM-04 in primary cortical neurons treated with L-glutamate, oligomeric Aβ, or H2O2. PSM-04 exhibited significant neuroprotective effects against neurotoxicity induced by L-glutamate or oligomeric Aβ was studied. PSM-04 exhibited significant neuroprotective effects against neurotoxicity induced by L-glutamate or oligomeric Aβ. Oxidative stress induced by ROS was monitored using the DCF-DA assay, and apoptosis was assessed using the TUNEL assay in primary cortical neurons treated with H2O2 or oligomeric Aβ. PSM-04 also decreased oxidative stress induced by H2O2 and apoptotic cell death induced by oligomeric Aβ. We evaluated the therapeutic effect of PSM-04 in 5xFAD (Tg) mice, an animal model for AD. PSM-04 was orally administered to 4-month-old 5xFAD mice for 2 months. To confirm the degree of cognitive impairment, a novel object recognition task was performed. The treatment with PSM-04 significantly alleviated cognitive impairments in Tg mice. In addition, amyloid plaques and gliosis decreased significantly in the brains of PSM-04-administered Tg mice compared with Tg-vehicle mice. Furthermore, the administration of PSM-04 increased the superoxide dismutase-2 (SOD-2) protein level in hippocampal brain tissues. Our results indicated that PSM-04 showed therapeutic effects by alleviating cognitive impairment and decreasing amyloid plaque deposition in Tg mice. Therefore, PSM-04 was considered as a potential pharmacological agent for neuroprotective effects in neurodegenerative diseases, including AD.
    Keywords:  5xFAD mice; Alzheimer’s disease; PSM-04; Polygala tenuifolia Willdenow; neuroprotection
    DOI:  https://doi.org/10.3389/fphar.2019.00002
  46. J Pathol. 2019 Feb 08.
    Herrington CS, Poulsom R, Coates PJ.
      In this Annual Review Issue of The Journal of Pathology, we present 15 invited reviews on topical aspects of pathology, ranging from the impacts of the microbiome in human disease through mechanisms of cell death and autophagy to recent advances in immunity and the uses of genomics for understanding, classifying and treating human cancers. Each of the reviews is authored by experts in their fields and our intention is to provide comprehensive updates in specific areas of pathology in which there has been considerable recent progress.
    Keywords:  ARDS; Breast Cancer Index; CD44; DAMP; DNA damage response; EndoPredict (EPclin); Epstein-Barr virus; FoxO3A; GATA3; ILC2; KRT5; Lynch syndrome; MLKL; MammaPrint; MapQuantDx; Melanoma; NanoString; OncotypeDX; PAMP; PD-1; PD-L1; PPARgamma; Prosigna; RB1; TGFbeta; TP53; VUS; abscopal response; acral; acute kidney injury, transplantation; adaptive immunity; alveolarisation; apoptosis; autoimmunity; autophagy; basal-like bladder cancer; biobank; bioinformatics; biomarkers; breast cancer; bronchopulmonary dysplasia; cancer genes; cancer predisposition; cancer stem cell; cancer syndromes; caspase-3; checkpoint inhibitors; chemokine; chemoprevention; chronic inflammation; circulating tumour DNA (ctDNA); cutaneous; desmoplastic; dormancy; driver genes; drug resistance; dysbiosis; extracellular matrix; familial adenomatous polyposis; ferroptosis; fibroblasts; gasdermin; genomic signatures; group 2 innate lymphoid cells; hedgehog; hereditary colorectal cancer; hypoxia; imaging mass cytometry; immune-oncology; in situ hybridisation; inflammation; innate immunity; interferon; interleukin; leukotriene; liquid biopsy; lung cancer, small cell lung cancer, pathology, molecular genetics, in vitro models; lung development; lymphatic system; macrophages; massively parallel sequencing; matrix metalloproteinase; metastasis; microbiota; mismatch repair, microsatellite instability; mitochondria; mucosal; multiplexed immunohistochemistry; multiplexed ion beam imaging; muscle-invasive bladder cancer; mutations; myeloid-derived suppressor cells; nasopharyngeal carcinoma; necroptosis; neutrophil extracellular traps; neutrophils; polyposis; population health science; postnatal development; predictive; prognostic; prognostic signatures; protease; pyroptosis; quiescence; radiation; reactive oxygen species; respiratory infection; screening; severe inflammatory response syndrome; size-based diagnostics; statistics; stem cells; tissue biomarkers; toll-like receptor; translational research; triple negative breast cancer; tumour invasion; tumour microenvironment; tumour progression; tumour-infiltrating lymphocytes; type 2 immunity; urothelial carcinoma; uveal; variants of uncertain significance
    DOI:  https://doi.org/10.1002/path.5255
  47. Nefrologia. 2019 Jan 31. pii: S0211-6995(19)30004-9. [Epub ahead of print]
    Kumaş M, Eşrefoğlu M, Karataş E, Duymaç N, Kanbay S, Ergün IS, Üyüklü M, Koçyiğit A.
      BACKGROUND: Ischemia-reperfusion injury causes various severe morphological and functional changes in diabetic patients. To date, numerous antidiabetic and antioxidant agents have been used for treatment of the disease-related changes.OBJECTIVES: We aimed to examine effective therapeutic doses or doses of berberine against renal ischemia/reperfusion injury (IRI) in a streptozotocin (STZ)-induced diabetic rat model by histopathological and biochemical analysis.
    METHODS: Thirty male Sprague Dawley rats were treated with STZ injection for the development of diabetes, and divided into the following groups: STZ-induced diabetic group (STZ); IRI-induced diabetic group (STZ+IRI); 50mg/kg berberine (BRB) treated diabetic group after inducing IRI (STZ+IRI+BRB1); 100mg/kg BRB treated diabetic group after IRI (STZ+IRI+BRB2); 150mg/kg BRB treated diabetic group after IRI (STZ+IRI+BRB3). Bilateral renal ischemia model was applied for 45min, then reperfusion was allowed for 14 days in STZ-induced diabetic rats. Renal injury was detected histopathologically. Blood urea nitrogen (BUN), creatinine and lactate dehydrogenase (LDH) levels were measured in serum using the ELISA method. Total antioxidant status (TAS) and total oxidant status (TOS) of renal tissue was studied by spectrophotometric assay. Oxidative stress index (OSI) was calculated as TOS-to-TAS ratio. Tumor necrosis factor alpha (TNF-α), C-reactive protein (CRP), Na+/K+-ATPase (sodium pump), and Ca2+-ATPase (calcium ATPase) enzyme levels were measured in tissues using the ELISA method. Anti-apoptotic Bax and pro-apoptotic Bcl-2 protein levels were detected by Western blot analysis. All data were evaluated statistically.
    RESULTS: The highest histopathological score was detected in the STZ+IRI group compared to the other group. BRB administration at the doses of 100mg/kg and 150mg/kg markedly improved renal injury. BUN and creatinine levels significantly increased in the STZ+IRI group compared to the STZ group (p<0.001). 100mg/kg and 150mg/kg BRB administration significantly decreased those levels (p<0.01). The highest TOS and the lowest TAS levels were detected in the STZ+IRI group (p<0.001). IRI markedly aggravated inflammation via increasing levels of TNF-α and CRP (<0.001), and caused apoptosis via inducing Bcl-2 protein, and suppressing Bax protein (p<0.001). BRB administration at the doses of 100mg/kg and 150mg/kg showed anti-oxidant, anti-inflammatory and anti-apoptotic effects (p<0.01). The LDH enzyme, was used as a necrosis marker, was higher in the STZ+IRI group than other groups. BRB administration at all of the doses, resulted in the decline of LDH enzyme level (p<0.001). Ca2+-ATPase and Na+/K+-ATPase enzyme activities decreased in the STZ+IRI group compared to the STZ group (p<0.001), while BRB administration at the doses of 100mg/kg and 150mg/kg significantly increased those of enzyme activities, respectively (p<0.05).
    CONCLUSION: Ischemia with diabetes caused severe histopathological and biochemical damage in renal tissue. The high doses of berberine markedly improved histopathological findings, regulated kidney function via decreasing BUN and creatinine levels, and rearranged intercellular ion concentration via increasing Na+/K+-ATPase and Ca2+- ATPase levels. Berberine showed anti-oxidant, anti-apoptotic, and anti-inflammatory effects. According to these data, we suggest that berberine at the doses of 100 and 150mg may be used as a potential therapeutic agent to prevent renal ischemic injury.
    Keywords:  Apoptosis; Berberina; Berberine; Estrés oxidativo; Histopathology; Histopatología; Inflamación; Inflammation; Insuficiencia renal por isquemia/reperfusión; Oxidative stress; Renal ischemia/reperfusion injury
    DOI:  https://doi.org/10.1016/j.nefro.2018.10.006
  48. PLoS One. 2019 ;14(2): e0208399
    Pearsall EA, Cheng R, Matsuzaki S, Zhou K, Ding L, Ahn B, Kinter M, Humphries KM, Quiambao AB, Farjo RA, Ma JX.
      Diabetic retinopathy (DR) is a common neurovascular complication of type 1 diabetes. Current therapeutics target neovascularization characteristic of end-stage disease, but are associated with significant adverse effects. Targeting early events of DR such as neurodegeneration may lead to safer and more effective approaches to treatment. Two independent prospective clinical trials unexpectedly identified that the PPARα agonist fenofibrate had unprecedented therapeutic effects in DR, but gave little insight into the physiological and molecular mechanisms of action. The objective of the present study was to evaluate potential neuroprotective effects of PPARα in DR, and subsequently to identify the responsible mechanism of action. Here we reveal that activation of PPARα had a robust protective effect on retinal function as shown by Optokinetic tracking in a rat model of type 1 diabetes, and also decreased retinal cell death, as demonstrated by a DNA fragmentation ELISA. Further, PPARα ablation exacerbated diabetes-induced decline of visual function as demonstrated by ERG analysis. We further found that PPARα improved mitochondrial efficiency in DR, and decreased ROS production and cell death in cultured retinal neurons. Oxidative stress biomarkers were elevated in diabetic Pparα-/- mice, suggesting increased oxidative stress. Mitochondrially mediated apoptosis and oxidative stress secondary to mitochondrial dysfunction contribute to neurodegeneration in DR. Taken together, these findings identify a robust neuroprotective effect for PPARα in DR, which may be due to improved mitochondrial function and subsequent alleviation of energetic deficits, oxidative stress and mitochondrially mediated apoptosis.
    DOI:  https://doi.org/10.1371/journal.pone.0208399
  49. ACS Appl Mater Interfaces. 2019 Feb 08.
    Zhang L, Yi H, Song J, Huang J, Yang K, Tan B, Wang D, Yang N, Wang ZG, Li X.
      Sonodynamic therapy (SDT), a promising alternative for cancer therapy, utilizes a sonosensitizer combined with ultrasound (US) irradiation to damage tumor cells/tissue for therapeutic purposes. The ability for sonosensitizers to specifically accumulate in tumor cells/tissues could greatly influence their therapeutic efficiency. In this work, we report the use of US-activated sonosensitizer (IR780)-based nanodroplets (IR780-NDs) for SDT, which provide numerous benefits for killing cancer cells compared to traditional methods. For instance, IR780-NDs showed effective surface to core diffusion both in vitro and in vivo. In the presence of US, the acoustic droplet vaporization (ADV) effect significantly assisted the conveyance of IR780-NDs from the circulatory system to tumor regions, and the acoustic wave force also increased the penetration depth within tumor tissues. Furthermore, IR780-NDs possesses mitochondrial targeting capabilities, which improves the precision and accuracy of SDT delivery. During in vitro assessment, the overproduction of reactive oxygen species (ROS) was observed following mitochondrial targeting, which rendered cancer cells more susceptible to ROS-induced apoptosis. Additionally, IR780-ND is a suitable candidate for photoacoustic (PA) and fluorescence (FL) imaging, and can also enhance US imaging due to the ADV-generated bubbles, which provides the potential for SDT guidance and monitoring. Therefore, with combined modalities, IR780-NDs can be a promising theranostics nanoplatform for cancer therapy.
    DOI:  https://doi.org/10.1021/acsami.8b21968
  50. J Photochem Photobiol B. 2019 Jan 15. pii: S1011-1344(18)31356-3. [Epub ahead of print]192 90-96
    Elshawi OE, Nabeel AI.
      Radiotherapy is considered as a primary modality for cancer treatment which accompanied by several side effects. Protection of normal tissues from radiation effects is one of the most significant concerns for researchers. Although many compounds acting as radio protectors, only two compounds were licensed clinically. Cyclooxygenase-2 (COX-2), as an inflammatory mediator is associated with ROS production with a NF-κB gene up regulation dependent manner in normal tissues. To that extend, his study was designed to target COX-2 and NF-κB by a newly synthesized benzopyran-4-one or chromone derivative; (2E)-2-((4-oxo-4H-chromen-3-yl) methylene amino-4- nitrobenzoic acid (Ch). Exposure of mice to IRR significantly induced intestinal inflammation via overexpression of COX-2 and NF-κB which is accompanied by an increase in the levels of MDA and iNOS in tissue homogenate and in the production of TNF-α and IL-6 as inflammatory signs. Moreover, the apoptotic effect of IRR was manifested by obvious elevation in caspase-3. Interapretonial injection of Ch significantly controls the inflammatory response by blocking the COX-2 and decrease the expression NF-κB which subsequently decreases other inflammatory parameters. Thus Ch compound might be a promising nonsteroidal anti-inflammatory drug (NSAID) against radiation-induced inflammation with a specific mode of COX-2 inhibition. Further researches are needed to elucidate its molecular mechanism and its combination with radiotherapy as a protector.
    Keywords:  Apoptosis; Chromone derivative; Inflammation; Oxidative stress; Radiotherapy
    DOI:  https://doi.org/10.1016/j.jphotobiol.2019.01.006
  51. Chemosphere. 2019 Jan 24. pii: S0045-6535(19)30147-X. [Epub ahead of print]222 355-363
    Cao X, Wang S, Bi R, Tian S, Huo Y, Liu J.
      Hexavalent chromium [Cr(VI)] is the main harmful component in the atmosphere released by chemical industry. The study was conducted to assess Cr(VI) inducing cardiovascular diseases (CVDs) in vitro by investigating the effects of Cr(VI) on bovine hemoglobin (BHb) and human umbilical vein endothelial cells (HUVECs). Multi-spectroscopic techniques and molecular docking method were used to determine the interaction of Cr(VI) and BHb. Fluorescence spectra results showed that the quenching constant (Ksv) decreased with temperature raise, indicating that Cr(VI) quenches BHb fluorescence through static quenching mechanism. The number of binding sites was 1.14 (310 K), enthalpy and entropy changes revealed the interaction of Cr(VI) and BHb was driven by hydrogen bonds. The results of synchronous fluorescence and circular dichroism (CD) spectra suggested that Cr(VI) could change BHb conformation and influence the microenvironment of Trp and Tyr residues. Moreover, in order to study Cr(VI) induced HUVECs damage, inflammatory factors were detected at the mRNA level, JNK and p38 MAPK pathways were analyzed. The results shown that Cr(VI) could induce mRNA expression of NLRP3, ICAM-1, VCAM-1, TNF-α and IL-1β, and increased intracellular ROS. Furthermore, Cr(VI) could induce oxidative stress in HUVECs, and then activate JNK and p38 MAPK pathways, ultimately lead to apoptosis of HUVECs by activating mitochondrial apoptosis pathways. These results suggested that Cr(VI) might bring about CVDs by both changing the BHb conformation and inducing HUVECs damage.
    Keywords:  Cr(VI); Fluorescence; HUVECs; Inflammatory; Oxidative stress
    DOI:  https://doi.org/10.1016/j.chemosphere.2019.01.137
  52. Toxicol Res (Camb). 2019 Jan 01. 8(1): 67-76
    Li M, Wang S, Li X, Kou R, Wang Q, Wang X, Zhao N, Zeng T, Xie K.
      The purpose of the present study was to investigate the effects and underlying mechanisms of diallyl sulfide (DAS), an organosulfur compound extracted from garlic, on drug-induced or chemical-induced liver injury caused by acetaminophen (APAP) or carbon tetrachloride (CCl4) in mice. DAS (100, 200, or 400 μmol kg-1) was orally administered 1 hour before APAP or CCl4 intraperitoneal injection, and the serum and liver tissue were collected 24 hours after APAP or CCl4 exposure. The serum aminotransferase activities and liver histopathological examination showed that DAS exhibited obvious hepatoprotective effects against acute liver injury induced by APAP or CCl4. In addition, exposure to APAP or CCl4 resulted in an increased content of malonaldehyde as well as a decreased ratio of reduced to oxidized glutathione, and a decreased level of superoxide dismutase and catalase activity in the liver (p < 0.05); however, pretreatment with DAS restored the perturbations of the antioxidant system in the liver. Beyond that, DAS pretreatment reduced the APAP-/CCl4-induced increase in phosphorylation of inhibitor of kappa B alpha (IκBα) and p65 subunit of nuclear factor kappa B (NF-κB) expression in the cytoplasm and nucleus in the liver. DAS pretreatment also decreased the excessive level of TNF-α caused by APAP or CCl4 in serum (p < 0.05). Moreover, DAS pretreatment regulated the expression of cleaved caspase 3, Bax and Bcl-2 in the liver and suppressed APAP-/CCl4-induced hepatocyte apoptosis. In conclusion, DAS exhibits hepatoprotective effects against drug-induced and chemical-induced liver injuries induced by APAP or CCl4 in mice, probably due to its ability to reduce hepatic oxidative stress and inhibit inflammatory injury and hepatocyte apoptosis.
    DOI:  https://doi.org/10.1039/c8tx00185e
  53. Oxid Med Cell Longev. 2019 ;2019 9529676
    Wang PW, Cheng YC, Hung YC, Lee CH, Fang JY, Li WT, Wu YR, Pan TL.
      Extensive exposure to UVB (280-320 nm) is the major risk responsible for various skin injuries. Numerous reports have shown that natural products could demonstrate photochemopreventive efficacy against UVB damage. We investigated the preventive effects and associated molecular mechanisms of red raspberry extract upon UVB-caused damage in human epidermal keratinocytes and a nude mouse model. The protein profiles and immunohistological study on a nude mouse skin indicated that red raspberry extract could prevent UVB-caused cell death and protect the skin against UVB-exposed injury manifested by wrinkling, scaling, tanning, and water loss as well as epidermal thickening. In addition, red raspberry extract application effectively abolished oxidative damage in DNA and attenuated the carbonylation level of proteins, which attributed to the activation of SOD, Nrf2 and its target genes, and HO-1. Red raspberry extract also altered the cells' apoptotic signaling pathways including caspase-3 as well as the inflammatory cascade such as c-jun and attenuated UVB-induced activation of NF-κB and COX-2. Red raspberry extract could alleviate direct photodamage to the skin caused by UVB exposure through the ROS scavenger and protection against inflammatory responses, which may allow the development of novel strategies in protecting the skin subjected to UVB radiation.
    DOI:  https://doi.org/10.1155/2019/9529676
  54. Oxid Med Cell Longev. 2019 ;2019 7670854
    Yu LM, Dong X, Zhang J, Li Z, Xue XD, Wu HJ, Yang ZL, Yang Y, Wang HS.
      Endoplasmic reticulum (ER) stress and oxidative stress contribute greatly to myocardial ischemia-reperfusion (MI/R) injury. Naringenin, a flavonoid derived from the citrus genus, exerts cardioprotective effects. However, the effects of naringenin on ER stress as well as oxidative stress under MI/R condition and the detailed mechanisms remain poorly defined. This study investigated the protective effect of naringenin on MI/R-injured heart with a focus on cyclic guanosine monophosphate- (cGMP-) dependent protein kinase (PKG) signaling. Sprague-Dawley rats were treated with naringenin (50 mg/kg/d) and subjected to MI/R surgery with or without KT5823 (2 mg/kg, a selective inhibitor of PKG) cotreatment. Cellular experiment was conducted on H9c2 cardiomyoblasts subjected to simulated ischemia-reperfusion treatment. Before the treatment, the cells were incubated with naringenin (80 μmol/L). PKGIα siRNA was employed to inhibit PKG signaling. Our in vivo and in vitro data showed that naringenin effectively improved heart function while it attenuated myocardial apoptosis and infarction. Furthermore, pretreatment with naringenin suppressed MI/R-induced oxidative stress as well as ER stress as evidenced by decreased superoxide generation, myocardial MDA level, gp91 phox expression, and phosphorylation of PERK, IRE1α, and EIF2α as well as reduced ATF6 and CHOP. Importantly, naringenin significantly activated myocardial cGMP-PKGIα signaling while inhibition of PKG signaling with KT5823 (in vivo) or siRNA (in vitro) not only abolished these actions but also blunted naringenin's inhibitory effects against oxidative stress and ER stress. In summary, our study demonstrates that naringenin treatment protects against MI/R injury by reducing oxidative stress and ER stress via cGMP-PKGIα signaling. Its cardioprotective effect deserves further clinical study.
    DOI:  https://doi.org/10.1155/2019/7670854
  55. J Trace Elem Med Biol. 2019 Mar;pii: S0946-672X(18)30479-6. [Epub ahead of print]52 74-82
    Salama SA, Arab HH, Hassan MH, Al Robaian MM, Maghrabi IA.
      Cadmium is an extremely toxic pollutant that reaches human body through intake of the industrially polluted food and water as well as through cigarette smoking and exposure to polluted air. Cadmium accumulates in different body organs especially the liver. It induces tissue injury largely through inflammation and oxidative stress-based mechanisms. The aim of the current study was to investigate the ability of γ glutamyl cysteine (γGC) to protect against cadmium-induced hepatocellular injury employing Wistar rats as a mammalian model. The results of the current work indicated that γGC upregulated the level of the anti-inflammatory cytokine IL-10 and downregulated the levels of the pro-inflammatory cytokines (TNF-α, IL-6, and IL-1β) in the cadmium-exposed rats. In addition, γGC reduced the liver tissues cadmium content in the cadmium-treated rats, suppressed the cadmium-induced hepatocellular apoptosis and oxidative modifications of cellular DNA, lipids, and proteins. Additionally, γGC enhanced the antioxidant potential of the liver tissues in the cadmium-treated rats as evidenced by a remarkable increase in the activity of the antioxidant enzymes superoxide dismutase and glutathione peroxidase and significant increase in the levels of the total antioxidant capacity and reduced glutathione as well as a significant reduction in oxidized to reduced glutathione (GSSG/GSH) ratio. Moreover, it effectively improved liver cell integrity in the cadmium-treated rats as demonstrated by a significant reduction in the serum activity of the liver enzymes (ALT and AST) and amelioration of the cadmium-evoked histopathological alterations. Together, these findings underscore, for the first time, the alleviating effects of γGC against cadmium-induced hepatocellular injury that is potentially mediated through reduction of liver tissue cadmium content along with modulation of both hepatocellular redox status and inflammatory cytokines.
    Keywords:  8-Oxo-2′- deoxyguanosine; Cadmium; Inflammatory cytokines; Liver; Oxidative stress
    DOI:  https://doi.org/10.1016/j.jtemb.2018.12.003
  56. Neurotoxicol Teratol. 2019 Jan 30. pii: S0892-0362(18)30133-8. [Epub ahead of print]
    Nasrallah G, Salem R, Da'as S, Al-Jamal OLA, Scott M, Mustafa I.
      OBJECTIVES: Clinically approved iron chelators are effective in decreasing significant transfusional iron accumulation. Starch-Deferoxamine (S-DFO), a novel high molecular weight iron chelator, was produced to increase binding capacity to iron and reduce toxicity. Although its efficacy was established in one small cohort clinical trial, its potential adverse effect was not adequately addressed.METHODS: We utilized zebrafish model to assess S-DFO toxicity using following assays: mortality, teratogenicity, hatching rate, tail flicking, Acridine Orange staining for apoptosis detection, o-dianisidine staining for hemoglobin synthesis, and the level of Hsp70 as a general stress indicator. Embryos were exposed to different concentrations of S-DFO, Zinc Oxide nanoparticle (ZnO) (positive control), along with untreated control (UC).
    RESULTS: S-DFO showed no significant mortality nor deformities at all tested concentrations (0.0-1000 μM). Thus, the LC50 is expected to >1000 μM. 100 μM S-DFO treatment did not affect embryo development (as judged by hatching rate); neuromuscular activity (as judged by tail flicking); and hemoglobin synthesis. Neither apoptosis, nor increase in Hsp70 level was noticed upon S-DFO treatment.
    CONCLUSION: Our assays demonstrate that S-DFO does not induce cellular or biochemical stress and has no adverse effect on organ development of zebrafish embryos, suggesting its safe use as an iron chelator.
    Keywords:  Iron chelation; S-DFO; Toxicity; Zebrafish; ZnO
    DOI:  https://doi.org/10.1016/j.ntt.2019.01.004
  57. J Dairy Res. 2019 Feb 07. 1-5
    Santillo A, Figliola L, Caroprese M, Marino R, dʼApolito M, Giardino I, Albenzio M.
      We evaluated the effect of in vitro digested milk on mature adipocytes 3T3-L1, paying particular attention to its fatty acid composition, and comparing human (HM), donkey (DM), bovine (BM), ovine (OM), caprine (CM) and formula (FM) milk. Cellular viability, apoptosis, oxidative response and gene expression levels of NF-κB p65, HMGB1, SREBP-1c and FAS were evaluated. Digested milk treatments significantly reduced 3T3-L1 mature adipocytes viability and caspase activity compared with control group, but no significant differences were observed among different sources of digested milk. In all digested milk samples, ROS level was higher than the control, however, the digested human and formula milk showed lower levels of ROS than DM, BM, OM and CM samples. Lower capacity of HM and FM to induce oxidative stress in mature adipocytes was ascribed to the peculiar free fatty acids profile of digested milk samples. All milk treatments elicited a significant over-expression of NF-κB p65 in 3T3-L1 adipocytes compared to the control; the lowest gene expression was found in HM, BM, OM and CM, the highest in FM and an intermediate behavior was shown in DM. All digested milk treatments influenced the gene expression of SRBP-1c with FM and HM showing the highest levels. For FAS expression, BM showed the highest level, OM and CM intermediate and FM, HM and DM the lowest levels, however HM and DM had comparable levels to the control.
    Keywords:  3T3-L1 adipocyte; digested milk; fatty acids; human milk; milking species
    DOI:  https://doi.org/10.1017/S0022029919000104
  58. Lasers Med Sci. 2019 Feb 02.
    Maldaner DR, Azzolin VF, Barbisan F, Mastela MH, Teixeira CF, Dihel A, Duarte T, Pellenz NL, Lemos LFC, Negretto CMU, da Cruz IBM, Duarte MMMF.
      Skin aging is a complex biological process induced by intrinsic and extrinsic factors which is characterized by clinical and cellular changes, especially dermal fibroblasts. It is possible that, some procedures, such as low-level laser therapy (LLLT), could decelerate this process. To test this hypothesis, this study evaluated the in vitro LLLT on dermal fibroblast cell line (HFF-1) with premature senescence H2O2-induced. HFF-1 cells were cultured in standardized conditions, and initially H2O2 exposed at different concentrations. Fibroblasts were also just exposed at different LLLT (660 nm) doses. From these curves, the lowest H2O2 concentration that induced indicators of premature senescence and the lowest LLLT doses that triggered fibroblast proliferation were used in all assays. Cellular mortality, proliferation, and the levels of oxidative, inflammatory cytokines, apoptotic markers, and of two growth signaling molecules (FGF-1 and KGF) were compared among treatments. The H2O2 at 50 μM concentration induced some fibroblast senescence markers and for LLLT, the best dose for treatment was 4 J (p < 0.001). The interaction between H2O2 at 50 μM and LLLT at 4 J showed partially reversion of the higher levels of DNA oxidation, CASP 3, CASP 8, IL-1B, IL-6, and INFy induced by H2O2 exposure. LLLT also trigger increase of IL-10 anti-inflammatory cytokine, FGF-1 and KGF levels. Cellular proliferation was also improved when fibroblasts treated with H2O2 were exposed to LLLT (p < 0.001). These results suggest that in fibroblast with some senescence characteristics H2O2-induced, the LLLT presented an important protective and proliferative action, reverting partially or totally negative effects triggering by H2O2.
    Keywords:  Aging; Fibroblast; Low-level laser; Skin
    DOI:  https://doi.org/10.1007/s10103-019-02728-1
  59. Mol Genet Genomic Med. 2019 Feb 03. e548
    Talepoor Ardakani M, Rostamian Delavar M, Baghi M, Nasr-Esfahani MH, Kiani-Esfahani A, Ghaedi K.
      BACKGROUND: Parkinson's disease (PD) is ranked as the second most common neurodegenerative disorder caused by loss of dopaminergic neurons in the substantia nigra. Micro(mi)RNAs are a class of small noncoding RNAs that regulate gene expression and aberrant expression of them is closely correlated with many neurodegenerative conditions including PD. Silent information regulator 1 (SIRT1) as a known deacetylase and B-cell lymphoma-2 (BCL2) as an antiapoptotic factor play vital roles in neural protection and survival.METHODS: Differentiated PC12 cells exposed to MPP+ were served here as a known PD model. Cell viability was determined by MTS assay. Apoptotic cells and ROS levels were detected using flow cytometry. Gene selection and miRNA-mRNA interaction analysis were performed through in silico methods. Relative expression of miRNAs and genes was examined by RT-qPCR.
    RESULTS: MPP+ exposure markedly reduced cell viability, enhanced oxidative stress, and induced apoptosis in differentiated PC12 cells. Sirt1 and BCL2were shown to be markedly declined in response to MPP+ , while miR-200a and miR-204 were significantly upregulated.
    CONCLUSION: The first novel finding of the current study is altered expression of miR-200a and miR-204 in differentiated PC12 cells in response to MPP+ , suggesting that deregulation of them participate in MPP+ neurotoxicity mechanisms, possibly via affecting the expression of Sirt1 and BCL2 as potential targets.
    Keywords:  MPP+; Parkinson’s disease; differentiated PC12 cell; miR-200a; miR-204
    DOI:  https://doi.org/10.1002/mgg3.548
  60. Hum Mol Genet. 2019 Jan 30.
    Hajji K, Mteyrek A, Sun J, Cassar M, Mezghani S, Leprince J, Vaudry D, Masmoudi-Kouki O, Birman S.
      Parkinson's disease (PD) is a progressive neurodegenerative movement disorder that can arise after long-term exposure to environmental oxidative stressors, such as the herbicide paraquat (PQ). Here we investigated the potential neuroprotective action of vertebrate pituitary adenylate cyclase-activating polypeptide (PACAP) against PQ in Drosophila. We found that pretreatment with this neuropeptide applied to the ventral nerve cord (VNC) at low doses markedly extended the survival of wild-type decapitated flies exposed to neurotoxic levels of PQ or dopamine (DA). In contrast and interestingly, application of a PACAP receptor antagonist, PACAP-6-38, had opposite effects, significantly decreasing the resistance of flies to PQ. PACAP also reduced PQ-induced caspase activation and reactive oxygen species (ROS) accumulation in the VNC. We then searched for the endogenous neuropeptide receptor potentially involved in PACAP-mediated neuroprotection in Drosophila. Knocking down the gene encoding the receptor Han/PDFR of the neuropeptide pigment-dispersing factor (PDF) in all neurons conferred to flies higher resistance to PQ, whereas PDFR downregulation restricted to PDF or DA neurons did not increase PQ resistance, but remarkably suppressed the neuroprotective action of PACAP. Further experiments performed with Pdf and Pdfr-deficient mutant strains confirmed that PDF and its receptor are required for PACAP-mediated neuroprotection in flies. We also provide evidence using split-GFP reconstitution that PDF neurons make synaptic contacts onto DA neurons in the abdominal VNC. Our results, therefore, suggest that the protective action of PACAP against PQ-induced defects in the Drosophila nervous system involves the modulation of PDFR signaling in a small number of interconnected neurons.
    DOI:  https://doi.org/10.1093/hmg/ddz031
  61. Front Pharmacol. 2018 ;9 1554
    Nemecz M, Constantin A, Dumitrescu M, Alexandru N, Filippi A, Tanko G, Georgescu A.
      In this study, we aimed to identify the mechanisms underlying the different effects of palmitic acid and oleic acid on human pancreatic beta cell function. To address this problem, the oxidative stress, endoplasmic reticulum stress, inflammation, apoptosis and their mediator molecules have been investigated in the insulin releasing beta cells exposed to palmitic and/or oleic acid. Herein, we have demonstrated that in cultured 1.1B4 beta cells oleic acid promotes neutral lipid accumulation and insulin secretion, whereas palmitic acid is poorly incorporated into triglyceride and it does not stimulate insulin secretion from human pancreatic islets at physiologically glucose concentrations. In addition, palmitic acid caused: (1) oxidative stress through a mechanism involving increases in ROS production and MMP-2 protein expression/gelatinolytic activity associated with down-regulation of SOD2 protein; (2) endoplasmic reticulum stress by up-regulation of chaperone BiP protein and unfolded protein response (UPR) transcription factors (eIF2α, ATF6, XBP1u proteins) and by PTP-1B down-regulation in both mRNA and protein levels; (3) inflammation through enhanced synthesis of proinflammatory cytokines (IL6, IL8 proteins); and (4) apoptosis by enforced proteic expression of CHOP multifunctional transcription factor. Oleic acid alone had opposite effects due to its different capacity of controlling these metabolic pathways, in particular by reduction of the ROS levels and MMP-2 activity, down-regulation of BiP, eIF2α, ATF6, XBP1u, CHOP, IL6, IL8 and by SOD2 and PTP-1B overexpression. The supplementation of saturated palmitic acid with the monounsaturated oleic acid reversed the negative effects of palmitic acid alone regulating insulin secretion from pancreatic beta cells through ROS, MMP-2, ATF6, XBP1u, IL8 reduction and SOD2, PTP-1B activation. Our findings have shown the protective action of oleic acid against palmitic acid on beta cell lipotoxicity through promotion of triglyceride accumulation and insulin secretion and regulation of some effector molecules involved in oxidative stress, endoplasmic reticulum stress, inflammation and apoptosis.
    Keywords:  endoplasmic reticulum stress; human pancreatic beta cells; inflammation; oleic acid; oxidative stress; palmitic acid
    DOI:  https://doi.org/10.3389/fphar.2018.01554