辛伐他汀保护缺血-再灌注损伤心肌的线粒体蛋白质组学研究  

Proteomics of Ischemia and Reperfusion Myocardial Mitochondria by Administration of Simvastatin

作　　者：陈学颖[1] 孙爱军[1,2] 弭守玲 樊慧芝[2,3] 徐磊 赵吉[1] 王克强[1] 邹云增[1,2] 葛均波[1,2] 

机构地区：[1]复旦大学附属中山医院上海市心血管病研究所,上海200032 [2]复旦大学生物医学研究院,上海200032 [3]复旦大学化学系,上海200433

出　　处：《中国分子心脏病学杂志》2010年第6期360-365,共6页Molecular Cardiology of China

基　　金：国家重点基础研究发展计划(973计划)(2011CB503905);国家高技术研究发展计划资助项目(863计划)(2006AA0ZA406);国家杰出青年科学基金(30725036);国家自然科学基金(30971250)

摘　　要：目的建立大鼠心肌缺血-再灌注损伤模型,通过蛋白质组学的方法研究辛伐他汀对大鼠缺血-再灌注损伤心肌线粒体代谢的保护作用。方法将大鼠随机分为辛伐他汀干预组(n=14)和生理盐水对照组(n=14),建立缺血-再灌注损伤模型,通过伊文思蓝和TTC染色评估梗死面积,提取大鼠左心室心肌线粒体蛋白行双向凝胶电泳,应用质谱分析鉴定差异蛋白点。结果辛伐他汀组和对照组相比,梗死区与危险区(梗死区+缺血区)的比值有统计学差异(29.4%±8.4%vs57.7%±6.5%,P<0.0001);梗死面积与左室面积的比值有统计学差异(15.9%±5.6%vs29.0%±8.9%,P=0.012)。双向凝胶电泳图谱分析有19个蛋白点的表达有差异,质谱鉴定了9种差异蛋白,相比对照组,辛伐他汀组4个蛋白表达上调:三功能酶亚基α(线粒体前体)、电子转移黄素蛋白脱氢酶、肌动蛋白α(心肌)、细胞色素c氧化酶亚基5A亚单位(线粒体前体);5个蛋白表达下调:L-乳酸脱氢酶B链、异柠檬酸脱氢酶[NAD]α亚基(线粒体前体)、α晶状体蛋白B链、内膜蛋白(线粒体)、肌动蛋白类似物(细胞质)。结论辛伐他汀组大鼠心肌在缺血-再灌注损伤后,心肌梗死面积显著减少,辛伐他汀改变线粒体呼吸链、能量代谢等途径上的蛋白,为阐明辛伐他汀保护缺血再灌注损伤提供了理论依据。Objective To identify the protective effect in rat myocardial mitochondria with simvastatin following myocardial ischemia and reperfusion injury by comparative proteomics.Methods Sprague-Dawley rats were randomized to receive either simvastatin(n=14)or saline(n=14)before myocardial ischemia and reperfusion.After reperfusion,six rats from each group were used to calculate the infarction ratio by Evans blue and triphenyltetrazolium chloride(TTC)staining.Total proteins extracted from the mitochondria were applied to the two-dimensional gel electrophoresis.The differentially expressed protein spots were evaluated by a software.Then they were subjected to in-gel digestion,and analyzed by thematrix-assisted laser desorptionionization time-of-flightmass spectrometry(MALDI-TOF-MS).Results The ratio of the infarct size and the risk area(ischemia area + infarct area)was 29.4%±8.4% in the simvastatin group and 57.7%±6.5% in controls(P < 0.0001).The ratio of the infarct size and the left ventricular myocardia was 15.9%±5.6% in the simvastatin group and 29.0%±8.9% in controls(P=0.012).Nineteen differentially expressed protein spots were observed,and nine of them were analyzed by MALDI-TOF-MS.Four proteins including trifunctional enzyme subunit alpha(mitochondrial precursor),electron-transferring-flavoprotein dehydrogenase,actin alpha(cardiac muscle 1)and cytochrome c oxidase subunit 5A(mitochondrial precursor)were up-regulated and five proteins including L-lactate dehydrogenase B chain,isocitrate dehydrogenase [NAD] subunit alpha(mitochondrial precursor),alpha crystallin B chain,inner membrane protein(mitochondrial)and similar to actin(cytoplasmic 2)exhibited down-regulation in the simvastatin group compared to the controls.Conclusions The infarct size of the myocardium in the simvastatin group was significantly reduced as compared to the controls.Several mitochondrial proteins concerning respiratory chain and energy metabolism in the simvastatin group were identified by proteomic analysis,which may probably illuminate th

关 键 词：辛伐他汀 缺血-再灌注损伤 线粒体 蛋白质组 

分 类 号：R96[医药卫生—药理学]