氯化钴对乳鼠心肌细胞凋亡的影响

目的确定萌化钴(CoCl2)对乳鼠心肌细胞凋亡的影响。方法采用达氏修正依氏(DMEM)/F12(1∶1)培养基原代培养乳鼠心肌细胞,3d后分别给予浓度200、400、600μmol/L的CoCl2培养24h,对照组不加CoCl2。锥虫蓝染色检测细胞存活力,Hochest33258荧光染色检测细胞凋亡,用Western Blot检测缺氧诱导因子-1α(HIF-1α)蛋白水平。结果与对照组比较,CoCl2培养组细胞存活率明显下降(P<0.01),凋亡率明显增加(P<0.01),且心肌细胞存活率下降程度及细胞凋亡率睁加程度均随着氯化钴浓度增加而升高。HIF-1α蛋白水平亦随CoCl2浓度升高而增加。结论CoCl2可模拟缺氧诱导乳鼠心肌细胞凋亡。

Effects of cobaltchloride on neonatal rat cardiac myocyte apoptosis

Objective To identify the effect of cobalt chloride(CoCl2)on cells apoptosis in primary neonatal rat cardiac myocytes.Methods Isolation and culturing the primary neonatal rat cardiac myocytes were performed in DMEM/F12(1∶1)medium.After 3 days,cells were incubated for 24 h in medium added CoCl2 with different concentration(200,400,600 μmol/L respectively).Controm group was free of CoCl2.Cell viability was measured by Trypan blue staining.Cardiomyocyte apoptotic rate was detected with Hoechst33258 DNA binding assay.Western blot was used to detect the protein level of hypoxia inducible factor 1α(HIF-1α).Results Compared with control group,myocyte survival rate significantly decreased(P<0.01),and apoptotic rate significantly increased in cells treated with CoCl2(P<0.01).Moreover,the extent of decreasing of cell survival rate and increasing of apoptotic rate were larger with increasing concentration of CoCl2.However,the protein level of hypoxia HIF-1α was also higher in accompany with increasing concentration of CoCl2.Conclusion CoCl2 can mimic hypoxia to induce apoptosis in cultured primary neonatal rat cardiac myocytes.