Hongkun Xue1, Qian Li2, Jiaqi Tan3, Pengcheng Li4,5. 1. Key Laboratory of Particle & Radiation Imaging, Ministry of Education, Department of Engineering Physics, Tsinghua University, No. 30 Shuangqing Road, Haidian District, Beijing, 100084, China. xuehk@mail.tsinghua.edu.cn. 2. Key Laboratory of Particle & Radiation Imaging, Ministry of Education, Department of Engineering Physics, Tsinghua University, No. 30 Shuangqing Road, Haidian District, Beijing, 100084, China. liqianld@126.com. 3. Academy for Advanced Interdisciplinary Studies, Peking University, No. 5 Yiheyuan Road, Haidian District, Beijing, 100871, China. 4. Key Laboratory of Particle & Radiation Imaging, Ministry of Education, Department of Engineering Physics, Tsinghua University, No. 30 Shuangqing Road, Haidian District, Beijing, 100084, China. 5. College of Food Science and Technology, Jilin Agricultural University, No. 2888 Xincheng Street, Changchun, 130118, Jilin, China.
Abstract
OBJECTIVE: The aim of this study is to evaluate the cytoprotection and potential molecular mechanisms of cyanidin-3-glucoside (C3G) on hydrogen peroxide (H2O2)-induced oxidative damage in HepG2 cells. METHODS: The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay was carried out to examine the viability of HepG2 cells exposure to H2O2 or C3G. Meanwhile, the antioxidant properties of C3G were measured by determining the activities of superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), catalase (CAT) and the malondialdehyde (MDA) levels. Flow cytometry was employed to determine HepG2 cells apoptosis, and HepG2 cells were stained with Hoechst 33342 to observe cell morphology. 2',7'-dichlorofluorescin diacetate (DCFH-DA) was used to evaluate the production of intracellular reactive oxygen species (ROS). Finally, the expression of apoptosis-related protein was monitored through western blot analysis. RESULTS: HepG2 cells induced with H2O2 presented a remarkable decrease in cell viability that was suppressed when HepG2 cells were interfered with C3G (2.5-10 μM). C3G interference memorably and dose-dependently inhibited H2O2-induced intracellular ROS and MDA overproduction, while C3G treatment markedly increased H2O2-induced the activities of intracellular SOD, GSH-Px and CAT. Eventually, the relative proteins expression levels of p53, cleaved caspase-9/3, cytochrome c, Fas-L, Fas, FADD and caspase-8 were substantially up-regulated in H2O2-triggered HepG2 cells, and Bax/Bcl-2 ratio and the relative protein expression levels of PARP were dramatically down-regulated. However, the expression levels of these relative proteins were reversed in C3G-interfered HepG2 cells. CONCLUSIONS: C3G could protect HepG2 cells from oxidative damage, and the effects that were mediated by the mitochondrial apoptotic pathways and the external pathways.
OBJECTIVE: The aim of this study is to evaluate the cytoprotection and potential molecular mechanisms of cyanidin-3-glucoside (C3G) on hydrogen peroxide (H2O2)-induced oxidative damage in HepG2 cells. METHODS: The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay was carried out to examine the viability of HepG2 cells exposure to H2O2 or C3G. Meanwhile, the antioxidant properties of C3G were measured by determining the activities of superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), catalase (CAT) and the malondialdehyde (MDA) levels. Flow cytometry was employed to determine HepG2 cells apoptosis, and HepG2 cells were stained with Hoechst 33342 to observe cell morphology. 2',7'-dichlorofluorescin diacetate (DCFH-DA) was used to evaluate the production of intracellular reactive oxygen species (ROS). Finally, the expression of apoptosis-related protein was monitored through western blot analysis. RESULTS:HepG2 cells induced with H2O2 presented a remarkable decrease in cell viability that was suppressed when HepG2 cells were interfered with C3G (2.5-10 μM). C3G interference memorably and dose-dependently inhibited H2O2-induced intracellular ROS and MDA overproduction, while C3G treatment markedly increased H2O2-induced the activities of intracellular SOD, GSH-Px and CAT. Eventually, the relative proteins expression levels of p53, cleaved caspase-9/3, cytochrome c, Fas-L, Fas, FADD and caspase-8 were substantially up-regulated in H2O2-triggered HepG2 cells, and Bax/Bcl-2 ratio and the relative protein expression levels of PARP were dramatically down-regulated. However, the expression levels of these relative proteins were reversed in C3G-interfered HepG2 cells. CONCLUSIONS: C3G could protect HepG2 cells from oxidative damage, and the effects that were mediated by the mitochondrial apoptotic pathways and the external pathways.
Authors: Linsey E S de Groot; Yanaika S Sabogal Piñeros; Suzanne M Bal; Marianne A van de Pol; Jörg Hamann; Peter J Sterk; Wim Kulik; René Lutter Journal: Clin Exp Allergy Date: 2019-04-09 Impact factor: 5.018
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