H_2O_2通过钙稳态失调诱导小鼠胚胎肝细胞凋亡  

H_2O_2 Induced Hepatocyte Apoptosis by Intracellular Calcium Homeostasis

作　　者：赵培[1] 闫晓风[2] 王晓玲[2] 

机构地区：[1]上海中医药大学教学实验中心,上海201203 [2]上海中医药大学基础医学院生物教研室,上海201203

出　　处：《中国细胞生物学学报》2015年第9期1257-1262,共6页Chinese Journal of Cell Biology

基　　金：上海市教委预算内项目(批准号:2014YSN22;2014YSN03);上海中医药大学教师学术共同体项目(批准号:p304030105)资助的课题~~

摘　　要：该文研究了体外培养肝细胞内钙离子浓度改变对细胞存活率、凋亡和增殖的影响。建立了H2O2诱导小鼠胚胎肝细胞损伤模型,CCK-8检测细胞存活率,Fura-2/AM负载检测细胞内[Ca2+]i;免疫荧光和Western blot分别检测STIM1和Orai1在细胞内的定位和含量;流式细胞术检测细胞凋亡;Brdu掺入检测细胞增殖。结果显示,H2O2刺激后细胞存活率降低为对照组的73%,凋亡细胞比例增加,增殖细胞数目显著减少,细胞内[Ca2+]i升高,STIM1和Orai1蛋白质水平增加,且STIM1可与Orai1蛋白质共定位。2-APB预处理组可以降低细胞内[Ca2+]i,减少STIM1和Orai1蛋白质表达水平,抑制STIM1和Orai1蛋白质的相互作用。结果表明,H2O2可通过影响细胞内钙离子稳态导致细胞凋亡。This work was aim to investigate the effect of intracellular calcium homeostasis on apoptosis in hepatocytes. H202 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 STIM 1 and Orai I were detected by immunofluorescence. The expressions of STIM 1, Orail, Bax were evaluated by Western blot. Apoptosis was assayed with flow cytometry. Our results indicated that after exposure to H202, Cell viability reduced to 73%, the number of apoptotic cells increased but proliferating cells reduced. Compared with the control, the intracellular [Ca2＋]i in H202-treated cells was significantly increased, protein expression of STIM1 and Orail was up-regulated, colocalization of STIM1 and Orail was increased; while the 2-APB treatment could reduce [ca2＋]i, down-regulate protein expression of STIM1 and Orail and decrease the colocalization of STIM1 and Orail. Our study suggested that H202 could induce cell apoptosis by cellular calcium homeostasis.

关 键 词：肝细胞 H2O2 钙离子稳态 细胞凋亡 

分 类 号：R575[医药卫生—消化系统]