早期心肌缺血大鼠心肌中BNP的时序性表达  

Temporal expression of BNP in rats with early myocardial ischemia

作　　者：翟利琴 郭相杰[1] 靳茜茜[1] 刘明哲[1] 郭华林 高彩荣 ZHAI Liqin;GUO Xiangjie;JIN Qianqian;LIU Mingzhe;GUO Hualin;GAO Cairong(Department of Pathology,Forensic School of Shanxi Medical University,Taiyuan 030001,China;Department of Pathology,Shanxi Provincial People’s Hospital)

机构地区：[1]山西医科大学法医学院病理教研室,太原030001 [2]山西省人民医院病理科

出　　处：《山西医科大学学报》2020年第3期224-230,共7页Journal of Shanxi Medical University

基　　金：山西省自然科学基金资助项目(201601D102070);山西省归国留学人员基金项目资助(2015-050)。

摘　　要：目的观察大鼠缺血心肌组织中BNP mRNA及蛋白时序性表达。方法在GEO数据库中检索到大鼠急性心肌缺血基因表达谱数据GSE4648,分析并设计实验验证。66只10-12周龄雄性SD大鼠分为对照组(n=6)、过敏性猝死组(n=6)、假手术组(n=6)及早期心肌缺血组(n=48)。早期心肌缺血组结扎左冠状动脉前降支造成急性心肌缺血模型,分别于造模成功后0.5,1,2,4,8,12,24,48 h脱臼处死大鼠,取左心室前壁缺血区心肌。假手术组缝合线穿过左心室前壁心肌组织,而不结扎冠状动脉,脱臼处死大鼠,取左心室前壁心肌组织。过敏性猝死组大鼠尾静脉注射鸡卵清蛋白诱导过敏性猝死模型,猝死后取左心室前壁心肌组织。对照组常规饲养1周后脱臼处死,取左心室前壁心肌。应用qRT-PCR、Western blot及免疫组化法检测所取心肌BNP mRNA及蛋白表达情况。结果GSE4648基因表达谱数据显示大鼠心肌缺血4 h时荧光信号强度开始升高,缺血24 h荧光信号达到最强,缺血48 h荧光信号开始减弱。验证实验显示,与对照组相比,大鼠心肌缺血2 h时,缺血区心肌BNP mRNA开始升高(P<0.05),并于缺血24 h时达到峰值(P<0.01),缺血48 h时下降,但仍高于对照组(P<0.01)。与对照组相比,大鼠心肌缺血4 h时,BNP蛋白表达开始上升(P<0.05),在缺血12 h和24 h时达到最高峰(P<0.01),缺血48 h时下降,但仍高于对照组(P<0.05)。大鼠心肌缺血4 h时,缺血区心肌免疫组化出现弱阳性(P<0.05),之后逐渐增强,缺血24 h时达到强阳性(P<0.01),缺血48 h阳性减弱,但仍高于对照组(P<0.05)。对照组、假手术组及过敏性猝死组三组间心肌BNP mRNA及蛋白表达水平差异无统计学意义(P>0.05)。结论早期心肌缺血时存在BNP mRNA升高;检测BNP mRNA可能对鉴别冠状动脉性猝死和过敏性休克死亡有帮助。Objective To observe the expression pattern of BNP mRNA and protein in ischemic myocardial tissue of rats at different time points.Methods Gene expression profile of acute myocardial ischemia in rats,GSE4648,was retrieved from GEO database and then analyzed.After that,confirmation experiment was carried out.Sixty-six male Sprague-Dawley rats of 10-12 weeks old were divided into control group(n=6),allergy-induced sudden death group(n=6),sham group(n=6)and early myocardial ischemia(EMI)group(n=48).The left anterior descending coronary artery in EMI group was ligated to induce myocardial ischemia,and then the rats were sacrificed at 0.5,1,2,4,8,12,24,48 h after the modeling.Rats in sham group underwent exactly the same procedure as ischemia group,except that the anterior descending artery was left open,and then were sacrificed.Rats in allergy-induced death group were injected with chicken ovalbumin to induce anaphylaxis and death.Rats in control group were routinely fed for one week and then were sacrificed.All hearts in each group were harvested.The mRNA and protein expression levels of BNP in each group were detected by qRT-PCR,Western blot and immunohistochemistry,respectively.Results GSE4648 data showed that BNP fluorescence intensity increased at 4 h after ischemia,and peaked at 24 h,and then decreased at 48 h after ischemia.The qRT-PCR demonstrated that,compared with control group,BNP mRNA in ischemic myocardium rose at 2 h after ischemia in EMI group(P<0.05)and peaked at 24 h(P<0.01),and then decreased at 48 h(P<0.05).Western blot showed that,compared with control group,BNP protein began to increase at 4 h after ischemia in EMI group(P<0.05),and peaked at 12 h and 24 h(P<0.01),and then decreased at 48 h(P<0.05).Immunohistochemistry demonstrated that,compared with control group,BNP immunoreactive score slightly increased in ischemic myocardium at 4 h after ischemia in EMI group(P<0.05),then gradually increased,and peaked at 24 h(P<0.01),and then decreased at 48 h(P<0.05).BNP mRNA and protein showed no change i

关 键 词：脑钠尿肽 心肌缺血 过敏性猝死 

分 类 号：DF795.4[医药卫生—法医学]