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《Chinese Journal of Cellular and Molecular Immunology》 2015-01
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Oxidative stress promotes hepatocyte apoptosis mediated by glycogen synthase kinase 3β

ZHANG Xiangying;GUO Yuanyuan;ZHANG Li;WEN Tao;PIAO Zhengfu;SHI Hongbo;CHEN Dexi;DUAN Zhongping;REN Feng;Beijing Youan Hospital,Capital Medical University,Beijing Institute of Hepatology;  
Objective To analyze the role of glycogen synthase kinase 3β(GSK3β) in hepatocyte apoptosis induced by oxidative stress.Methods Human HL-7702 hepatoma cells were induced by H2O2/antimycin A to establish oxidative stress-induced cell apoptosis models.SB216763,a specific inhibitor of GSK3β,was given to the cells two hours before H2O2/antimycin A induction.Cell survival was observed using calcein acetoxymethyl ester/propidium iodide(PI) double staining,and cell apoptosis was detected using annexin Ⅴ-FITC/PI staining combined with flow cytometry.In the meanwhile,the cel culture supernatant was subjected to lactate dehydrogenase(LDH) assay to evaluate the extent of cel death.The expressions of p-GSK3β,GSK3β,caspase-3,cleaved caspase-3,c-Jun N-terminal kinase(JNK) and cytochrome C(Cyt C)proteins were examined using Western blotting.Results Oxidative stress triggered by H2O2/antimycin A promoted GSK3β activity;inhibition of GSK3β activity by SB216763 relieved oxidative stress and reduced cell apoptosis induced by oxidative stress.Compared with the model groups,SB216763 intervened group showed that the cell apoptosis rate and the level of LDH were reduced significantly,and that the expressions of cleaved caspase-3,JNK,Cyt C proteins decreased.Conclusion GSK3β is an important signaling molecule in the apoptosis pathway induced by oxidative stress.The inhibition on GSK3β may alleviate the oxidative stress-induced hepatocyte apoptosis.
【Fund】: 国家125科技重大专项(2012ZX10002004-006 2012ZX10004904-003-001 2013ZX10002002-006-001);; 国家自然科学基金(81270532);; 北京市优秀人才培养资助(D类 2011D003034000022);; 2012北京市留学人员科技活动择优资助;; 首都临床特色应用研究(Z121107001012167);; 王宝恩肝纤维化研究基金(CFHPC20131031);; 北京市肝病研究所所内基金(BJIH-01403)
【CateGory Index】: R575.3
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