过氧化氢诱导C_2C_(12)细胞凋亡的机制初探  

Preliminary Investigation on the Mechanism of C_2C_(12) Cell Apoptosis induced by Hydrogen Peroxide

作　　者：肖卫民[1] 蒋碧梅[1] 石永忠[1] 刘梅冬[1] 唐道林[1] 肖献忠[1] 

机构地区：[1]中南大学湘雅医学院病理生理学教研室,湖南长沙410078

出　　处：《医学临床研究》2004年第11期1288-1291,共4页Journal of Clinical Research

基　　金：国家自然科学基金 (3 0 0 0 0 0 69;3 0 2 70 5 3 3 );国家 973重点项目 (G2 0 0 0 0 5 690 8);教育部博士点专项基金 (2 0 0 2 0 5 3 3 0 3 2 )资助

摘　　要：【目的】探讨氧化应激诱导小鼠胚胎肌原细胞株C2 C12 细胞凋亡的分子机制。【方法】采用 0 .5mmol/L过氧化氢 (hydrogenperoxide,H2 O2 )作用于C2 C12 细胞 ;通过Hoechst荧光染色检测C2 C12 细胞凋亡 ,Caspase活性定量分析及Western blot检测Caspase 3, 8, 9活化情况 ,Western blot及间接免疫荧光检测细胞色素C在细胞内的分布情况。【结果】本实验发现H2 O2 能明显诱导C2 C12 细胞凋亡 ,同时Caspase 3, 8, 9被激活 ,而细胞色素C从线粒体释放入胞浆。【结论】氧化应激可通过同时激活线粒体通路与死亡受体通路导致C2 C12 细胞凋亡 。ObjectiveTo investigate the molecular mechanism that apoptosis of mouse embryonic myogenic cell line C 2C 12 cells was induced by oxidative stress.Apoptosis of mouse C 2C 12 cells was induced by exposure to 0.5 mmol/L hydrogen peroxide (H 2O 2) for different durations, and their apoptotic changes were detected by Hoechst 33258 fluorescence staining. The activities of caspase -3, -8, -9 were examined by caspase colorimetric assay kit and Western-blotting. The intracellular distribution of cytochrome C and its release from mitochondria were observed by indirect immunofluorescence and Western-blotting.Exposure to 0.5 mmol/L H 2O 2 for 24 hours markedly induced C 2C 12 cell apoptosis as shown by Hoechst 33258 fluorescence staining. The activities of caspase -3, -8, -9 significantly increased after 4 hours of H 2O 2 treatment, and reached their peaks at 8～12 hour. The release of cytochrome C from mitochondria to cytoplasm was detected after exposure to H 2O 2 for 1～2 hours.[Conclusions]Oxidative stress can induce apoptosis of C 2C 12 cells through simultaneous activation of mitochondria and death receptor signal pathways. This result provides new information for clinical prophylaxis and treatment of apoptosis related cardiovascular diseases.

关 键 词：脱噬作用 过氧化氢 小鼠 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]