钾、氯离子通道阻断剂对staurosporine诱导心肌细胞凋亡的调节作用

Role of potassium and chloride channel blockers in apoptosis of cardiomyocytes induced by staurosporine

目的 :探讨钾、氯离子通道在心肌细胞凋亡过程中的调节作用与Caspase 3激活的关系 .方法 :在staurosporine诱导原代培养鼠心肌细胞凋亡模型中 ,观察钾、氯离子通道阻断剂 (Quinine ,BaCl2 ,DIDS和NPPB)对心肌细胞的存活率、DNA片断、Caspase 3活性及细胞膜完整性的影响 .实验分为阴性对照组 (无药物处理 )、阳性对照组 (staurosporine处理 )与药物干预组 (staurosporine加离子通道阻断剂 ) .结果 :①钾、氯离子通道阻断剂能有效的抑制staurosporine诱导的心肌细胞凋亡 .与阳性对照组 (2 9.8% )相比细胞的成活率在奎宁、氯化钡、DIDS和NPPB组分别是 5 9.7% ,4 7.2 % ,5 8.6 %和 6 0 .7% ,均有显著的增加 (P <0 .0 1 ) .相对应的DNA片断电泳显示 :Qui nine,BaCl2 ,DIDS和NPPB组均有显著的抑制DNA的降解 .②钾、氯离子通道阻断剂能有效的抑制Caspase 3活性的激活 ,与阳性对照组 (6 0 0 .4 % )相比细胞的Caspase 3活性在Quinine,BaCl2 ,DIDS和NPPB组分别是 1 84 .2 % ,2 1 6 .3% ,1 75 .6 %和2 2 1 .4 % ,均有显著的抑制 (P <0 .0 1 ) .③细胞膜完整性实验乳酸脱氢酶水平显示各组中细胞的坏死性死亡小于 1 0 % .结论 :在staurosporine诱导的心肌细胞凋亡模型中 ,钾、氯离子通道阻断剂能有效的抑制Caspase 3?

AIM: To explore the role of potassium and chloride channel blockers in cardiomyocyte apoptosis process and their relationship with caspase 3 activation. METHODS: Primarily cultured mouse cardiomyocytes were exposed to staurosporine and the cell viability, DNA fragmentations, caspase 3 activation, and cell membrane integrity were observed in three groups: Normal control (without treatment), positive control (staurosporine) and drug group (staurosporine + potassium or chloride channel blocker). RESULTS: ① Potassium and chloride channel blockers potently inhibited cardiomyocytes apoptosis induced by staurosporine. Compared to the positive control group, the cell viability in drug group increased significantly ( P <0.01) but apoptotic DNA fragmentations were suppressed in cardiomyocytes apoptosis induced by staurosporine. ② Potassium and chloride channel blockers also remarkably inhibited caspase 3 activity activated by staurosporine. Compared to the positive control group, caspase 3 activation decreased significantly ( P <0.01) in drug group. ③ In both positive control and drug groups, the cell lactate dehydrogenase (LDH) leakage was less than 10%. CONCLUSION: Potassium and chloride channel blockers can remarkably inhibit cardiomyocytes apoptosis mediated by caspase 3 activation in the model of mouse cardiomyocyte apoptosis induced by staurosporine.