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参附注射液对大鼠心肌缺血再灌注细胞凋亡的影响
引用本文:郑曙云,徐建国,李明.参附注射液对大鼠心肌缺血再灌注细胞凋亡的影响[J].南京大学学报(自然科学版),2004,40(2):192-198.
作者姓名:郑曙云  徐建国  李明
作者单位:南京大学医学院临床学院南京军区南京总医院麻醉科,南京大学医学院临床学院南京军区南京总医院麻醉科,中国药科大学心血管药理教研室 南京,210002,南京,210002,南京,210008
摘    要:通过观察参附注射液对缺血再灌注心肌超微结构和细胞凋亡参数的影响 ,探讨参附注射液拮抗心肌缺血再灌注损伤的作用机制 .使用结扎 /松解左冠状动脉前降支缺血 60min,再灌注 2 4 0min复制缺血再灌注动物模型 .Sprague_Dawley(SD)大鼠 40只随机分为 5组 :对照组(Ⅰ组 ,n =8) ,缺血再灌注组(Ⅱ组 ,1 /R组 ,n =8) ,参附组 (Ⅲ组 ,n=8) ,红参组 (Ⅳ组 ,n =8) ,附子组 (Ⅴ组 ,n =8) .Ⅲ、Ⅳ、Ⅴ各治疗组在缺血前 1 0min经静脉分别注射参附注射液 (1 0mg/kg)、红参注射液 (9mg/kg)、附子注射液 (1mg/kg) ,Ⅰ组、Ⅱ组在缺血前 1 0min经静脉注射同等容积生理盐水 .测定心肌组织丙二醛 (MDA)和超氧化物歧化酶 (SOD)值 ;电镜观察心肌细胞超微结构变化 ;TUNEL法原位标记凋亡心肌细胞 ,免疫组化和图像分析技术检测心肌细胞内Bcl_2、Bax蛋白表达 .与Ⅱ组比较 ,Ⅲ、Ⅳ、Ⅴ各治疗组 ,心肌组织MDA明显降低 ,有极显著差异(P <0 .0 1 ) .III组与IV组、V组之间比较 ,MDA降低 ,有显著性差异 (P <0 .0 5) .SOD活性明显升高 ,有显著差异 (P <0 .0 5) .心肌超微结构病变明显改善、减轻 .心肌细胞凋亡指数和Bax蛋白含量显著减少(P <0 .0 1 ) ,Bcl_2蛋白含量显著增多 (P <0 .0 1 ) .参附注射液具有抗大鼠心肌缺血再灌注损伤作用 ,

关 键 词:参附注射液  大鼠  心肌缺血  细胞凋亡  缺血再灌注损伤  Bcl-2  BaX  冠状动脉血运重建术

Effect of Shenfu Injection on Myocardial Apoptosis Induced by Ischemia/reperfusion Injury in Rat
Zheng Shu_Yun ,Xu Jian_Guo ,Li Ming.Effect of Shenfu Injection on Myocardial Apoptosis Induced by Ischemia/reperfusion Injury in Rat[J].Journal of Nanjing University: Nat Sci Ed,2004,40(2):192-198.
Authors:Zheng Shu_Yun  Xu Jian_Guo  Li Ming
Institution:Zheng Shu_Yun 1,Xu Jian_Guo 1,Li Ming 2
Abstract:To investigate the effects of shenfu injection,which was made of panax ginseng and aconitum carmichaeli,we observed the ultrastructure of myocardium and myocardial cell apoptosis and possible mechanism of myocardial protection affected by shenfu injection against ischemia/reperfusion(I/R)injury.The myocardial ischemia/reperfusion model of SD rat was established by using the method:ligating the left anterior descending branch of coronary artery for60minutes and then loosening it for240minutes.Forty SD rats were randomized into five groups:control group(I,n=8),with left anterior descending coronary artery exposed and a piece of silk thread placed around but not tied;I/R group(II,n=8),with normal saline(0.1mL/kg,i.v.)injected10min before I/R;shenfu injection group(III,n=8),with shenfu injection(10mg/kg,i.v.)injected10min before I/R;panax ginseng group(IV,n=8),with panax ginseng injection(9mg/kg,i.v)injected10min before I/R;aconitum carmichaeli group(V,n=8),with aconitum carmichaeli injection(1mg/kg,i.v.)injected10min before I/R.The animals were then sacrificed and the heart were harvested for determination of malondialdethyde(MDA)and superoxide dismutase(SOD)in myocardial tissue and ultrastructure examination(electron microscope).Apoptosis was identified by TUNEL and apoptosis index(AI)was obtained.The protein expression of Bax and Bcl_2genes was detected by using S.P immunohistochemical technique,and the mean optical density(OD)values of the positive fields of protein expression were quantitatively examined by using image analysis system.The results showed that MDA was significantly reduced and protested SOD activity in group III,IV and V.The pathological changes of myocardial ultrastructure in group III,IV and V showed relatively minor damage compared with that in group II(P<0.01).The AI in group III,IV and V was significantly higher than that of group I(P<0.01)while it was lower than that of group II(P<0.01).The protein expression of Bcl_2and Bax gene increased significantly in group II,III,IV and V compared with that of group I(P<0.05).Compared with that of group II,the OD value of Bcl_2protein and the protein expression ratio of Bcl_2/Bax increased significantly and the OD value of Bax protein decreased significantly in group III,IV and V(P<0.05).The present study suggested that shenfu injection could protect myocardium from ischemia/reperfusion injury by modulating the protein expression of Bcl_2and Bax genes and inhibit apoptosis following myocardial ischemia/reperfusion.
Keywords:shenfu injection  myocardial reperfusion injury  apoptosis  Bcl_2  Bax
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