凋亡通路蛋白表达对过氧化氢诱导PC12细胞凋亡的影响

Expression of Apoptosis Pathway Proteins Modulates H_2O_2-induced Apoptosis in PC12 Cells

目的探讨过氧化氢(H2O2)诱导PC12细胞凋亡的机制。方法 MTT测定细胞存活率,荧光染色观察细胞形态,透射电镜观察细胞的超微结构,流式细胞仪测定细胞凋亡率,Western blotting技术和流式细胞仪检测Bcl-2和Bax蛋白的表达,分光光度计测定Caspase-3释放率。结果 0.25 mM的H2O2可诱导PC12细胞凋亡,细胞的形态发生改变,细胞核受损,细胞内Bcl-2蛋白的表达降低、Bax蛋白的表达增加,Caspase-3被激活。结论 H2O2诱导了PC12细胞凋亡,其作用机制与细胞内Bcl-2、Bax蛋白的表达及Caspase-3活性密切相关。

Objective：To investigate the apoptotic effect of H2O2 in rat pheochromocytoma（PC12） cell line.Methods The cell viability was analyzed by MTT;the morphological changes were observed by Hoechst 33 258 staining analysis;the ultrathin sections of cells were observed by transmission electron microscopic（TEM）;the apoptosis rate,the protein of Bcl-2 and Bax were measured by the flow cytometry analysis（FCM and Western blotting）;the Caspase-3 activation was measuredusing Microplate Fluorometer.Results：H2O2（0.25 mM） could result in the decrease of cell viability,the increase of Bax and the dropping of Bcl-2 protein expression of PC12 cell.It increased the activity of Caspase-3 and the apoptotic rate of cells.Conclusions：The apoptosis in H2O2-induced PC12 cells was accompanied by down-regulation of Bcl-2,up-regulation of Bax and sequential activation of Caspase-3.