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H2O2 Induced Hepatocyte Apoptosis by Intracellular Calcium Homeostasis
Zhao Pei1, Yan Xiaofeng2, Wang Xiaoling2*
1Experiment Center for Teaching & Learning, Shanghai University of Traditional Chinese Medicine, Shanghai 201203, China;
2Department of Biology, School of Basic Medicine, Shanghai University of Traditional Chinese Medicine, Shanghai 201203, China
2Department of Biology, School of Basic Medicine, Shanghai University of Traditional Chinese Medicine, Shanghai 201203, China
Abstract: This work was aim to investigate the effect of intracellular calcium homeostasis on apoptosis in hepatocytes. H2O2 was used to induce hepatocyte injury， CCK-8 assay was used to measure cell viability. Cytosolic free calcium ion concentration was determined by Fura-2/AM. The cell proliferation and intracellular colocalization of STIM1 and Orai1 were detected by immunofluorescence. The expressions of STIM1， Orai1， Bax were evaluated by Western blot. Apoptosis was assayed with flow cytometry. Our results indicated that after exposure to H2O2， cell viability reduced to 73%， the number of apoptotic cells increased but proliferating cells reduced. Compared with the control， the intracellular [Ca2+]i in H2O2-treated cells was significantly increased， protein expression of STIM1 and Orai1 was up-regulated， colocalization of STIM1 and Orai1 was increased; while the 2-APB treatment could reduce [Ca2+]i， down-regulate protein expression of STIM1 and Orai1 and decrease the colocalization of STIM1 and Orai1.Our study suggested that H2O2 could induce cell apoptosis by cellular calcium homeostasis.
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