葛根素预处理对大鼠缺血再灌注心肌细胞凋亡的影响  

The impact of puerarin preconditioning on ischemia-reperfusion cadiocyte apoptosis of rats

作　　者：吴青[1] 周祖玉[2] 陶大昌[3] 

机构地区：[1]四川省卫生干部管理学院生理教研室,四川成都610041 [2]四川大学基础与法医学院生理教研室,四川成都610041 [3]四川大学华西医学院遗传研究室,四川成都610041

出　　处：《基础医学与临床》2005年第2期147-151,共5页Basic and Clinical Medicine

基　　金：四川省卫生厅科学研究课题 (0 30 10 4)

摘　　要：目的观察葛根素(Pur)预处理对缺血再灌注(I/R)心肌细胞凋亡的干预作用,并探讨其作用的可能机理。方法将原代培养的新生大鼠心肌细胞随机分为(1)正常对照组及正常+Pur组;(2 )模拟缺血组及缺血+Pur组;(3)模拟再灌注组及再灌注+Pur组。末端标记法(TUNEL法)检测心肌细胞凋亡,流式细胞仪检测细胞凋亡百分率,透射电镜观察心肌细胞凋亡的形态学特征、免疫组织化学法检测Bcl 2 /Bax蛋白表达,并检测胞内Ca2 + 浓度。结果(1)心肌细胞I/R后电镜观察到典型的凋亡超微结构特征;TUNEL染色可见心肌细胞凋亡阳性反应;免疫组化检测发现Bcl 2蛋白表达下调,Bax蛋白表达上调;胞内明显Ca2 + 超载。(2 )葛根素可显著降低I/R心肌细胞内Ca2 + 浓度、降低凋亡指数和凋亡百分率、上调Bcl 2蛋白表达、下调Bax蛋白表达。结论葛根素具有抗心肌细胞凋亡的作用,其机理可能与抑制胞内Ca2 + 超载,干预Bcl 2 /Bax蛋白表达有关。Objective To observ the intervention of puerarins preconditioning on Ischemia/Reperfusion (I/R) cadiocytes apoptosis and approaching its possible mechanism. Methods Primary cultured cadiocytes of neonatal rats were stochastically divided into (1)normal control group and normal+Pur group,(2)simulating ischemia group and ischemia+Pur group,(3) simulating reperfusion group and reperfusion+Pur group. The cadiocyte apoptosis was detected by the TdT-mediated dUTP nick end labeling(TUNEI) staining, the percentage of apoptosis was examined by the flow cytometer, the morphologic features of apoptotic cadiocyts were identified by EM microscopy, the expressions of Bcl-2/bax protein in apoptotic myocardium was observed. The consistency of Ca 2+ was examined too. In both ischemia and reperfasion groups, the typical ultrastructure features of apoptotic myocardium were observed by teransmission electron microscopic and the positive reaction of apoptotic cadiocytes was detected by the TdT-mediated dUTP nick end labeling(TUNEI)staining. Results The phenomena that Bc1-2 protein expression was in down regulation, but Bax protein expression was opposite were checked by means of immunohistochemistry and analysed by image inalysis system. The Ca 2+was overloading obviously in cells. Pur has the abilities of clearly reducing the consistency of Ca 2+in cadiocytes, reducing the index and the percentage of apoptosis too. It also can up-regulate the Bc1-2 protein expression and down-regulate the Bax protein expression. Conclusion Pur has the effect of resisting the cadiocytes apoptosis. Its possible mechanism could be that these is relationship with refraining the overloading Ca 2+in cells and interfering in the expression of Bcl-2/bax protein.

关 键 词：心肌细胞凋亡 葛根素 缺血再灌注 BCL-2蛋白表达 Bax蛋白 CA^2+超载 BCL-2/BAX 下调 上调 作用 

分 类 号：R285[医药卫生—中药学]