辛伐他汀对H2O2诱导心肌细胞凋亡抑制作用探究.docVIP

辛伐他汀对H2O2诱导心肌细胞凋亡抑制作用探究 　【摘要】 目的 研究辛伐他汀对H2O2诱导的心肌细胞凋亡的抑制作用并探讨其机制。方法 应用出生2～3 d的SD（Sprague-Dawley）大鼠乳鼠心肌细胞进行培养，用H2O2建立心肌细胞凋亡模型。随机分为五组：正常组，损伤组，1 μmol/L，3 μmol/L，10 μmol/L辛伐他汀保护组。损伤组H2O20.2 mmol/L作用6 h。甲基四唑蓝比色法（MTT法）测定心肌细胞活力；ＰＩ－ＡＶ双染后，流式细胞分析仪测定细胞凋亡率；用Western blot法测定caspase-3蛋白含量。结果 (1)H2O2能明显造成心肌细胞损伤:损伤组与正常组相比细胞活力下降。辛伐他汀1 μmol/L，3 μmol/L，10 μmol/L保护组对损伤心肌细胞有不同程度的保护作用(Plt;0.05)，并随剂量增加效果明显；（2）流式细胞仪检测细胞凋亡率损伤组比正常组增高(Plt;0.05)；辛伐他汀各保护组细胞凋亡率均比损伤组降低(Plt;0.05)，并随剂量增加效果明显；（3）用Western blot法测定caspase-3蛋白含量显示，损伤组比正常组caspase-3含量明显增加，辛伐他汀保护组可以降低H2O2引起的caspase-3蛋白含量增加，并随剂量增加效果明显。结论 辛伐他汀能够减轻H2O2引起的培养心肌细胞的凋亡,可能与抑制细胞内凋亡相关蛋白caspase-3有关。 【关键词】 辛伐他汀 心肌细胞 细胞凋亡 半胱氨酸天冬氨酸蛋白酶 过氧化氢 　　Abstract：Objective To investigate the protective effect and mechanism of simvastatin on cultured cardiomyocytes apoptosis by H2O2. Methods About 2～3 days old Sprague-Dawley rat neonatal cardiac myocytes were cultured, the cultured cells were divided into five groups, namely control group, damage group(H2O2 0.2 mmol/L), damage+1 mol/L simvastatin group, damage+3 mol/L simvastatin group, and damage+10 mol/L simvastatin group. The activity of dehydrogenase in mito-chondria was detected by MTT assay. Apoptosis rates of the cells were analyzed by flow cytometry. Expression of caspase-3 of cardiomyocytes were determined by Western blotting.Results (1) H2O2 could induce injury on cultured cardiac myocyte. This was evident in the descent of cell livability detected by MTT. (2)The rate of cell apoptosis of control group was 6.64%, the injury group was 24.28%, and different concentrations of simvastatin (1 mol/L,3 mol/L,10 mol/L) groups were 18.83%, 12.39%, 8.93%（Plt;0.01）.(3)Compared with the damage group expression of caspase-3, the simvastatin(1 mol/L,3 mol/L,10 mol/L) groups were much lower. Conclusions The results showed that simvastatin decreased the apoptosis of cardiomyocytes induced by H2O2, and the protective effect may be related to caspase-3. Key words：simvastatin; cardiomyocyte; apoptosis; caspase-3; H2