【摘要】 目的确定氯化钴(CoCl2)诱导PC12细胞凋亡的分子机制。方法500μmolLCoCl2诱导PC12细胞24h后,检测活性氧(ROS)生成量的变化以及抗氧化剂N乙酰半胱氨酸(NAC)、二硫代苏糖醇(DTT)对细胞存活率和ROS生成量的影响;琼脂糖凝胶电泳检测NAC、DTT对CoCl2诱导PC12细胞DNA片段化的影响;利用RTPCR法检测凋亡相关基因bclxl和bax在PC12细胞调亡过程中的表达及NAC、DTT对基因表达的影响;化学发光法检测NAC、DTT对Caspase3表达的作用。结果在CoCl2诱导PC12细胞凋亡中,ROS生成量明显上升,为正常时的2.92倍;NAC、DTT可提高细胞存活率,由53.1%上升为94.8%和92.3%(P<0.01),并有效抑制ROS的生成,为正常时的24.2%和82.1%(P<0.01);同时抑制DNA片段化的发生。bclxl在细胞凋亡过程中表达明显下降,bax表达无明显变化;NAC和DTT可使bclxl的表达明显上升。Caspase3在细胞凋亡中被激活,NAC、DTT可抑制Caspase3的表达(P<0.01)。结论ROS介导了CoCl2诱导的PC12细胞凋亡,这一过程与抑制bclxl表达,激活Caspase3有关。更多还原

【Abstract】 Objective To explore the mechanism of cobalt chloride（CoCl₂）-induced apoptosis in PC12 cells.Method The production of reactive oxygen species（ROS）in CoCl₂-induced apoptosis in PC12 cells was examined.The influence of NAC and DTT on cell viability,the production of ROS and the DNA fragment in PC12 cells were also examined.RT-CPR was used to examine the change of gene bcl-xl and bax expression in CoCl₂-induced apoptosis in PC12 cells and the effect of NAC and DTT on it.The expression of Caspase-3 and the influence of NAC and DTT on it were also examined.Results The production of ROS increased 2.92 times in CoCl₂-induced apoptosis in PC12 cells（P<0.01）.NAC and DTT improved the cell viability from 53.1% to 94.8% and 92.3%（P<0.01）.They could also inhibit the production of ROS to 24.2% and 82.1%（P<0.01） and DNA fragment.The expression of bcl-xl descended in the apoptosis,but bax had no change.Both NAC and DTT could protect the expression of bcl-xl.Caspase-3 was activated in the process,NAC and DTT could inhibit it （P<0.01）.Conclusion CoCl₂ induces PC12 cells apoptosis through ROS and accompanied with bcl-xl expression inhibited and Caspase-3 activated.更多还原