蛋白激酶C活化对大鼠缺血再灌注心肌细胞核三磷酸肌醇受体结合活性的影响  

Effect of protein kinase C activation on the binding activity of cardiac nuclear inositol 1,4,5-trisphosphate receptor in ischemia-reperfusion injured myocardium of rats

作　　者：张红[1] 司良毅[1] 周红[2] 张乐之[2] 何华美[2] 

机构地区：[1]解放军第三军医大学第一附属医院老年科,重庆市400038 [2]解放军第三军医大学药理学教研室,重庆市400038

出　　处：《中国临床康复》2005年第19期32-34,共3页Chinese Journal of Clinical Rehabilitation

基　　金：国家自然科学基金资助项目(30010710)~~

摘　　要：目的:观察具有抑制心肌细胞凋亡作用的蛋白激酶C活化对缺血再灌注心肌细胞核三磷酸肌醇受体结合活性的影响,以探讨其对心肌的保护作用。方法:实验于2003-04/07在解放军第三军医大学药理教研室完成。选用SD健康大鼠12只。随机分为2组,缺血再灌注组及假手术对照组,每组6只。缺血再灌注组:冠状动脉结扎30min,再灌注3h。假手术对照组:只埋线不结扎冠状动脉。采用蔗糖密度梯度离心法提纯心肌细胞核,将分离纯化的两组心肌细胞核分别平等进行2个亚组:磷酸化组和非磷酸化组,磷酸化组进行蛋白激酶C激动剂12-o-十四烷酰佛波醋酸酯-13磷酸化干预后,与非磷酸化组同时进行氚-1,4,5三磷酸肌醇的放射受体分析,比较两组细胞核经磷酸化后细胞核三磷酸肌醇受体结合活性变化。结果:进入结果分析大鼠12只。①心肌细胞凋亡指数:缺血再灌注组明显高于假手术对照组(9.88±0.33,0.6±0.0,P<0.01)。②大鼠心肌细胞核膜三磷酸肌醇受体的结合活性:缺血再灌注组与磷酸化组明显低于非磷酸化组犤(88.27±0.08),(136.05±0.05)pmol/g,P<0.05犦。③大鼠心肌细胞核膜三磷酸肌醇受体的结合力:磷酸化组和非磷酸化组无明显差异犤(102.83±0.09),(106.24±0.26)pmol/g,P>0.05犦。结论:大鼠心肌缺血再灌注时,蛋白激酶C可能是通过抑制心肌细胞核膜三磷酸肌醇受体的结合活性,进而参与了抑制缺血再灌注性心肌细胞凋亡的发生与发展。AIM:To observe the effect of protein kinase C,which can inhibit the myocardia l apoptosis,on the binding activity of cardiac nuclear inositol 1,4,5 trisphlo spate receptor of rat with myocardial ischemia reperfusion,so as to investigate its protective effect against myocardial ischemia reperfusion. METHODS:The experiment was carried out in the Department of Pharmacology,Thir d Military Medical University of Chinese PLA between April and July 2003.Twelve healthy SD rats were randomly divided into ischemia reperfusion group(n=6) and s ham operated control group(n=6).Ischemia reperfusion group:coronary artery was ligated for 30 minutes and reperfused for 3 hours;Sham operated control group :only embedded and coronary artery was not ligated.Myocardial nucleus was purifi ed with the method of saccharose density gradient centrifugation,and then the se parated and purified myocardial nucleus was divided into 2 subgroups respectivel y:phosphorylation and non phosphorylation groups.After the interventions of pr otein kinase C excitant 12 o tetradecanoylphorbol 13 acetate phosphoryla tion,rats in the phosphorylation group received the radioreceptor analysis of tr itium inositol 1,4,5 trisphlospate together with those in the non phosphoryl ation group,the changes of binding activity of nuclear inositol 1,4,5 trisphlo spate receptor after phosphorylation were compared between the two groups. RESULTS:All the 12 rats were involved in the analysis of results.① Myocardia l apoptosis index:It was obviously higher in the ischemia reperfusion group than in the sham operated control group(9.88± 0.33,0.6± 0.0,P< 0.01).② Cardiac nuclear binding activity of nuclear inositol 1,4,5 trisphlospate receptor:In t he ischemia reperfusion group,it was obviously lower in the phosphorylation subgroup than in the non phosphorylation subgr oup[(88.27± 0.08),(136.05± 0.05) pmol/g,P< 0.05].③ Cardiac nuclear binding ac tivity of nuclear inositol 1,4,5 trisphlospate receptor:In the sham operated control group,there was no obvious difference

关 键 词：心肌缺血 心肌再灌注损伤 蛋白激酶C 

分 类 号：R542.2[医药卫生—心血管疾病]