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• 基础研究 • 上一篇    下一篇

促凋亡蛋白Bid介导模拟缺血再灌注乳鼠心肌细胞的凋亡

于少娟, 赵学忠,张国成,纪 莉*   

  1. 吉林大学第一医院干部病房,吉林 长春130021
  • 收稿日期:2005-05-30 修回日期:1900-01-01 出版日期:2006-03-28 发布日期:2006-03-28
  • 通讯作者: 纪 莉*

Apoptosis of cardiocytes of neonatal rats with simulatedischemia/reperfusion injury mediated by pro-apoptosis by protein Bid

YU Shao-juan, ZHAO Xue-zhong,ZHANG Guo-cheng, JI Li*   

  1. Department of Cadre Ward,First Hospital, Jilin University, Changchun 130021,China
  • Received:2005-05-30 Revised:1900-01-01 Online:2006-03-28 Published:2006-03-28
  • Contact: JI Li*

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。

关键词: caspase-8, 再灌注损伤, 细胞凋亡, RNA干涉

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.

Key words: caspase-8, reperfusion injury, apoptosis, RNA interference

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