氯化钴对PC12细胞的凋亡作用

Apoptotic effect of cobalt chloride on PC12 cells

目的 确定氯化钴(CoCl2 )对PC12细胞的影响。方法 构建CoCl2 诱导的PC12细胞模型,检测CoCl2 对PC12细胞的毒性作用;将Caspases的抑制基因p3 5转染PC12细胞得到可稳定表达p3 5基因的细胞株PC12 p3 5 ,检测p3 5对CoCl2 诱导的PC12细胞的作用;检测Caspases特异性多肽抑制剂Z VAD FMK对CoCl2 诱导的PC12细胞的作用。结果 分别以10 0、3 0 0、5 0 0、70 0和10 0 0 μmol LCoCl2 诱导PC12细胞2 4h或以5 0 0 μmol LCoCl2 分别诱导PC12细胞12、2 4、3 6、48和60h后,PC12细胞存活率均明显下降(P <0 0 1) ,并与CoCl2 诱导的时间和浓度呈正相关;细胞亚显微结构检测,流式细胞分析和DNA片段化结果也表明5 0 0 μmol LCoCl2 诱导PC12细胞2 4h可使PC12细胞出现明显凋亡特征。构建可稳定表达Caspase蛋白酶抑制基因p3 5的PC12细胞株,或分别加入5 0和10 0 μmol LCaspases多肽抑制剂Z VAD FMK预处理PC12细胞1h后,以5 0 0 μmol LCoCl2 诱导PC12细胞2 4h ,形态学观察结果,细胞存活率检测和流式细胞分析均表明p3 5基因和Z VAD FMK均可有效地抑制CoCl2 诱导的PC12细胞凋亡(P <0 0 1)。结论 CoCl2 可诱导PC12细胞凋亡。

Objective To identify the effect of cobalt chloride (CoCl_2) on PC12 cells.Method The model that CoCl_2 induce apoptosis in PC12 cells was generated.PC12 cells that stable expressed the Caspases inhibitor gene p35 was generated and the effect of p35 and Z-VAD-FMK on the process of apoptosis induced by CoCl_2 was analyzed.Results PC12 cells triggered by 100,300,500,700 and 1 000 μmol CoCl_2 for 24 h or 500 μmol CoCl_2 for 12,24,36,48 and 60 h could both appear apoptosis in a dose and time-dependent manner (P<0.01). Electron microscope observation, flow cytometric analysis and DNA fragmentation could also identify the significant apoptosis in PC12 cells after induction by 500 μmol CoCl_2 for 24 h. The p35 gene and Z-VAD-FMK could significantly block the apoptosis induced by 500 μmol CoCl_2 for 24 h (P<0.01).Conclusion CoCl_2 can induce PC12 cells apoptosis.