【摘要】 采用不同浓度酒精处理PC12细胞,观察酒精诱导细胞凋亡作用及其与线粒体凋亡途径的关系.结果显示,采用50、100、200、400和800mmol/L浓度的酒精处理去血清培养的PC12细胞24h后,细胞存活率分别是单纯去血清培养细胞存活率的85.66%、74.28%、62.47%、54.14%和50.35%,呈浓度依赖性,与对照组比较差异显著(P<0.05).Hoechst 33342/PI染色法观察到典型凋亡现象,如核型固缩、染色质凝集等.DNA琼脂糖凝胶电泳可见100~400mmol/L浓度酒精处理组呈现凋亡细胞典型梯状DNA.酒精能使PC12细胞线粒体膜电位下降并呈浓度依赖性.酒精致PC12细胞凋亡过程中,凋亡关键蛋白Caspase-3、Caspase-9表达量升高.上述结果表明,酒精能够诱导PC12细胞凋亡,诱导作用随浓度升高而增强且酒精诱导PC12细胞凋亡与线粒体凋亡途径有关.更多还原

【Abstract】 In this study,the author established the ethanol-induced apoptotic model of pheochromocytoma(PC12)cells.Using the model,the author examined the inhibition effect of ethanol on proliferation of PC12 cells by MTT assay.Ethanol at the concentrations of 50,100,200,400 and 800mmol/L decreased cell proliferation of PC12 cells to 85.66%,74.28%,62.47%,54.14% and 50.35%,respectively,compared to the control in serum-free media after 24 h,indicating a dose-dependent inhibitory effect of ethanol on cell proliferation(P < 0.05).The nucleimorphological changes were detected by using Hoechst 33342 staining and fluorescent microscopy.Addition of ethanol with different concentrations to PC12 cells resulted in a number of morphological changes that are characteristic of apoptosis,including cell shrinkage,chromatin compaction and nuclear fragmentation.Agarose gel electrophoresis of the DNA isolated from ethanol(100~400 mmol/L)treated cells maintained in the absence of serum yielded the classical DNA laddering pattern of 180 to 200 bp fragments suggestive of apoptosis and a concomitant dissipation of the mitochondrial membrane potential.In addition,alcohol increased the protein expression of caspase-9and caspase-3.These results indicate that alcohol induced apoptosis of PC12 cells in a dosedependent manner,and mitochondrial-dependent pathways were involved in alcohol induction of apoptosis of PC12 cells.更多还原