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。
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 N AC and DTF 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 expressiun 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 ceils（ 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 bel-xl. Caspase-3 was activated in the process,NAC and DTT could inhibit it （P 〈 0.01 ）. Conclusion CoCl2 induces PCl2 cells apoptosis through ROS and accompanied with bcl-xl expression inhibited and Caspase-3 activated.
出处
《毒理学杂志》
CAS
CSCD
北大核心
2005年第3期193-195,共3页
Journal of Toxicology
基金
国家自然科学基金资助项目(30371564)
