活性氧在氯化锰致PC12细胞凋亡中的作用

目的 探讨活性氧（ROS）在氯化锰（MnCl2）诱导PC12细胞凋亡中的作用机制.方法 构建MnCl2诱导的PC12细胞模型,MTT法检测细胞存活率,流式细胞分析PC12细胞凋亡率;琼脂糖凝胶电泳检测DNA片段化程度;分光光度法检测培养基中活性氧（ROS）生成量变化;化学发光法检测细胞三磷酸腺苷（ATP）生成量和Caspase-3表达量的变化;RT-PCR法检测凋亡相关基因bcl-xl和bax的表达.结果 PC12细胞存活率与MnCl2诱导浓度和诱导时间呈负相关（P＜0.01）;2 mmol/L MnCl2诱导PC12细胞36 h,细胞凋亡率明显上升（P＜0.01）;核DNA发生明显片段化;细胞ROS生成量明显增加（P＜0.001）,细胞ATP生成量受到明显抑制（P＜0.01）;基因bcl-xl表达被抑制,bax表达上升（P＜0.01）;Caspase-3在细胞凋亡中被激活（P＜0.01）.结论 MnCl2诱导的PC12细胞凋亡与ROS升高、线粒体功能破坏和激活Caspase-3有关.

Mechanism of reactive oxygen species in manganese chloride-induced apoptosis in PC12 Cells

Objective To explore the mechanism of reactive oxygen species (ROS) in manganese chloride (MnCl_2)-induced apoptosis in PC12 cells. Methods The model that MnCl_2 induced apoptosis in PC12 cells was established.The apoptotic effect of MnCl_2 on PC12 cells was analyzed with the MTT,the flow cytometry and the DNA fragmentation.The production of ROS and ATP in MnCl_2-induced apoptosis of PC12 cells was examined.The influence of MnCl_2 on the expression of bcl-xl,bax and the activity of Caspase 3 was also analyzed. Results MnCl_2 triggered PC12 cells apoptosis in a dose-and time-dependent manner (P<0.01).The rate of aptosis was significantly increased (P<0.01) when MnCl_2 of 2 mmol/L induced PC12 cells for 36 hours.The production of ROS was increased (P<0.001) and the quantity of ATP was decreased (P<0.01) in PC12 cells with the same inducement of MnCl_2.The expression of bcl-xl was inhibited and the bax was activated in this process (P<0.01).Caspase 3 was also activated (P<0.01). Conclusion MnCl_2 induces apoptosis of PC12 cells,which is related to the increase of ROS,the inhibition of the mitochondria and the activation of Caspase3.