缝隙连接在大鼠缺血预适应心肌保护中的作用  

Role of gap junction in ischemic preconditioning

作　　者：苏德淳[1] 常志文[1] 范书英[2] 

机构地区：[1]首都医科大学附属北京同仁医院干部医疗科 [2]北京中日友好医院心内科

出　　处：《中华心血管病杂志》2006年第8期690-694,共5页Chinese Journal of Cardiology

基　　金：北京市教育委员会科技发展基金资助项目(KM200610025026)

摘　　要：目的探讨心肌缝隙连接在缺血预适应中的调控机制以及在预适应信号传导通路中的地位和作用。方法Sprague-Dawley大鼠进行30 min的冠状动脉缺血和4h再灌注,根据是否使用缺血预适应、二氮嗪、18β-甘草次酸和5-羟基奎酸分为7组,即对照组(单纯缺血再灌注)、缺血预适应组、缺血预适应和5-羟基奎酸合用组、二氮嗪组、二氮嗪和5-羟基奎酸合用组、甘草次酸组、甘草次酸和5-羟基奎酸合用组。测量各组的血流动力学指标、心肌梗死面积以及缝隙连接蛋白43的磷酸化和内化。结果与对照组相比,缺血预适应、二氮嗪、18β-甘草次酸能降低心肌梗死面积(尸< 0.001),5-羟基奎酸可以阻断缺血预适应和二氮嗪的心肌保护作用(P>0.05),但是对缝隙连接失耦联剂甘草次酸没有影响(P<0.001)。缺血预适应和线粒体三磷酸腺苷敏感性钾通道开放剂二氮嗪能促进缝隙连接蛋白43磷酸化,抑制去磷酸化和内化(P<0.001,P<0.001),线粒体三磷酸腺苷敏感性钾通道阻断剂5-羟基奎酸能阻断该作用(P>0.05,P>0.005 1)。结论缺血预适应通过开放线粒体三磷酸腺苷敏感性钾通道来调节缝隙连接蛋白43的磷酸化和内化,进而调控缝隙连接的化学转运,减少心肌梗死面积。Objective To investigate the role of gap junction in ischemic preconditioning （ IPC ）. Methods Sprague-Dawley rats were subjected to a 30 min coronary artery occlusion followed by 4 h of reperfusion （I/R）. Rats were divided into seven groups： I/R, IPC/R, IPC/R ＋ 5-hydroxydecanoic acid （mitochondrial ATP sensitive potassium channel antagonist） , I/R ＋ diazoxide （mitochondrial ATP sensitive potassium channel agonist）, I/R ＋ 5-hydroxydecanoic acid ＋ diazoxide, I/R ＋ 18β-glycyrrhetinic acid （gap junction blocker） and I/R ＋ 18β-glycyrrhetinic acid ＋ 5-hydroxydecanoic acid. Hemodynamics and myocardial infarct size were measured and connexin43 phosphorylation and subcellular distribution were determined by quantitative immunoblotting and confocal immunofluorescence. Results Infarct size was reduced in IPC/R, I/R ＋ diazoxide and I/R ＋ 18β-glycyrrhetinic acid group （ 13.34%±7. 87%, 11.02% ± 2. 24%, and 15.03% ± 11.35%, respectively; P 〈 0.001 vs. I/R group： 45. 81% ± 7.91%） . 5-hydroxydecanoic acid abolished the cardioprotective effects of IPC and diazoxide （46. 57% ± 5.36% and 47.36% ±3.17% ; P〉0.05 vs. I/R） but not the effects of glycyrrhetinic acid （14.60%±7.36% ; P〈 0. 001 vs. I/R）. Phosphorylation of connexin43 was significantly increased, dephosphorylation and connexin43 intracellular redistribution significantly decreased （ Cx43 size in the cellular membrane 1.00% ± 0.35% and 0.83%±0.31%, P〈0.001 vs. I/R： 0.19%±0.06%） by IPC and diazoxide and these effects could be abolished by 5-hydroxydecanoic acid. Conclusion Ischemic preconditioning could reduce myocardial infarction size by activating mitochondrial ATP sensitive potassium channel and modulating connexin43 phosphorylation and internalization.

关 键 词：再灌注损伤 心肌 连接蛋白 缺血预适应 

分 类 号：R541[医药卫生—心血管疾病]