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目的:直接暴露细胞于活性氧能诱导发生凋亡,本文研究氧化应激诱导HepG2肝癌细胞的死亡及其机制。方法:暴露细胞于2mmol/L过氧化氢产生氧化应激,用DNA凝胶电泳检测细胞凋亡,用荧光染色法检测细胞线粒体膜电位变化,Western blotting检测细胞浆中细胞色素c变化,fluorometric assay kit检测caspase活性变化。结果:氧化应激作用于HepG2细胞后12h开始发生凋亡;氧化应激作用后4h,细胞线粒体膜电位明显下降;胞浆中细胞色素c浓度呈时间依赖性增高;氧化应激作用8h、12h后细胞内caspase-3、caspase-9活性分别升高6.7及3.6倍,但caspase-8活性无变化。结论:氧化应激能诱导HepG2肝癌细胞发生凋亡,其途径与线粒体通路及caspase激活有关。
Aims: Direct exposure of cells to reactive oxygen species (ROS) can induce apoptosis. This study was designed to investigate the mechanism of oxidative stress-induced apoptosis in HepG2 hepatocarcinoma cells. Methods: The cells were exposed to 2mmol / L hydrogen peroxide to induce oxidative stress. The apoptosis of cells was detected by DNA gel electrophoresis. The changes of mitochondrial membrane potential were detected by fluorescence staining. The cytochrome c changes were detected by Western blotting. Fluorometric assay kit assay caspase activity changes. Results: Oxidative stress began to occur at 12h after HepG2 cells apoptosis. At 4h after oxidative stress, the mitochondrial membrane potential decreased markedly. Cytochrome c concentration in cytoplasm increased in a time-dependent manner. Oxidative stress was induced by 8h and 12h After intracellular caspase-3, caspase-9 activity increased by 6.7 and 3.6 times, respectively, but no change in caspase-8 activity. Conclusion: Oxidative stress can induce apoptosis in HepG2 hepatocarcinoma cells, which is related to mitochondrial pathway and caspase activation.