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1～3d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2h，复氧4h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。
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 ischernia/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.
出处
《吉林大学学报（医学版）》
CAS
CSCD
北大核心
2006年第2期271-274,共4页
Journal of Jilin University：Medicine Edition
基金
吉林省科技厅资助课题(990581-03)
