【摘要】 目的:探讨促凋亡蛋白Bid在模拟缺血再灌注乳鼠心肌细胞凋亡中的作用及其可能的机制。方法:培养出生1~3 d的SD乳鼠心肌细胞,随机分为正常对照组、模拟缺血再灌注组、模拟缺血再灌注+无关siRNA组、模拟缺血再灌注+Bid特异siRNA组。各组心肌细胞经缺氧2 h,复氧4 h建立缺血再灌注损伤模型;脂质体转染法(Oligofectamine)转入Bid无关的siRNA及Bid特异的siRNA,转入Bid特异的siRNA以拆除Bid基因的表达;免疫印迹技术(Western blotting)检测心肌细胞中Bid及caspase-8的活化;流式细胞术(flow cytometry,FCM)检测心肌细胞的凋亡率。结果:模拟缺血再灌注时心肌细胞Bid及caspase-8被活化。转染Bid特异siRNA组的心肌细胞caspase-8的活化较模拟缺血再灌注组及转染无关siRNA组均明显减少,心肌细胞凋亡率(7.4%)较模拟缺血再灌注组(51.2%)及转染无关siRNA组(48.7%)明显降低(P<0.01),而与正常对照组凋亡率接近(4.6%)。结论:促凋亡蛋白Bid介导了模拟缺血再灌注时心肌细胞的凋亡。更多还原

【Abstract】 Objective To study the role of Bid in simulated ischemia/reperfusion injury in cultured neonatal rat cardiocytes and its possible mechanism.Methods A cell culture model of neonatal rat(born within 3 days) cardiocytes was used to establish simulated ischemia/reperfusion model.Unrelated siRNA or Bid specific siRNA was transfected into rat cardiocytes by Oligofectamine.The activation of Bid and caspase-8 was detected by Western blotting.The apoptotic rate of cardiocytes was determined by flow cytometry.Results Bid and caspase-8 of cardiocytes were activatied in simulated ischemia/reperfusion injury model.The activation of caspase-8 of cardiocytes in Bid-specific siRNA group decreased significantly compared with those in simulated(ischemia/reperfusion) group and unrelated siRNA group;the apoptotic rate of cardrocytes(7.4%) was lower than those in simulated ischemia/reperfusion group(51.2%) and unrelated siRNA group(48.7%)(P<0.01),and approached the rate in control group(4.6%).Conclusion Bid mediates the apoptosis of rat cardiocytes in simulated(ischemia/reperfusion) injury.更多还原