线粒体凋亡途径在硫酸黏菌素致PC12细胞凋亡中的作用

Effect of mitochondria mediated pathway of apoptosis induced by colistin sulfate in PC12 cells

探讨线粒体凋亡途径在硫酸黏菌素诱导PC12细胞凋亡中作用。取对数生长期PC12细胞,用含0、62.5、125、250μg.mL-1硫酸黏菌素DMEM培养液,作用24 h,采用Hoechst33258荧光染色法检测PC12细胞凋亡,Western blot法检测细胞色素C(Cyt-C)、Bax、Bcl-2蛋白表达含量,试剂盒检测Caspase-3活性。与对照组相比,125、250μg.mL-1硫酸黏菌素剂量组细胞PC12细胞Cyt-C、Bax蛋白表达量、Caspase-3活性显著升高(P<0.01);Bcl-2蛋白表达显著降低(P<0.01),具有剂量-效应关系。说明线粒体途径介导PC12细胞凋亡是硫酸黏菌素神经毒性作用机制之一。

In order to explore the effects of mitochondria mediated pathway of colistin sulfate induced apoptosis in PC12 cells. PC12 cells were treated with colistin sulfate dissolved in the DMEM for 24 h, whose final concentrations were 0, 62.5,125 and 250μg. mL-1, respectively. The apoptosis was detected by Hoechst 33258 fluorescent staining. Expression of Cytochrome C （Cyt-C）, Bax, Bcl-2 protein was detected by westem blotting. The activities of Caspase-3 were examined by Kit. Compared with control group, 125 and 250 μg . mL-1 colistin sulfate could significantly increase （P〈0.01） the expression of Cytochrome C, Bax protein, the activity of caspase-3 in PC12 cell, significantly reduce （P〈0.01） the expression of Bcl-2 protein. These trends had dose-effect relationship. The results showed that the apoptosis induced by mitochondria pathway was one of neurotoxicity mechanisms caused by colistin sulfate.