重组腺病毒介导MRTF-A基因干预缺血再灌注损伤模型大鼠心肌细胞的凋亡  

Recombinant adenovirus encoding myocardin-related transcription factor-A prevents cardiomyocyte apoptosis induced by ischemia/reperfusion injury in rats

作　　者：杨辉 牛美芝 郭晓丽 刘福垒 Yang Hui;Niu Mei-zhi;Guo Xiao-li;Liu Fu-lei(Feicheng Mining Center Hospital,Feicheng 271608,Shandong Province,China;Central Hospital of Taian City,Taian 271000,Shandong Province,China)

机构地区：[1]山东省肥城矿业中心医院,山东省肥城市271608 [2]山东省泰安市中心医院,山东省泰安市271000

出　　处：《中国组织工程研究》2018年第24期3843-3848,共6页Chinese Journal of Tissue Engineering Research

基　　金：山东省自然科学基金(ZR2014HL108)~~

摘　　要：背景:有研究表明上调myocardin相关转录因子A(myocardin-related transcription factor-A,MRTF-A)基因表达可抑制脑缺血再灌注诱导的神经细胞凋亡而发挥脑保护作用,但其对心肌缺血-再灌注损伤中心肌细胞的作用鲜有报道。目的:探讨重组腺病毒介导MRTF-A基因对大鼠缺血再灌注损伤心肌细胞凋亡的影响。方法:SD大鼠随机分为假手术组、模型组、空白载体组和Adv-MRTF-A组,制备MRTF-A cDNA重组腺病毒载体,除假手术组外构建大鼠缺血再灌注损伤模型。Adv-MRTF-A组取大鼠左心室游离壁上、下、左、右4点各注射病毒滴度1.2×10^(10) TU/mL的Adv-MRTF-A;空白载体组以同样的方法注射同量的空白对照病毒,假手术组、模型组不注射。14 d后取材,TTC染色法检测各组大鼠心肌梗死面积,TUNEL法检测各组大鼠心肌细胞凋亡,Western blot法检测各组大鼠心肌组织中MRTF-A、Bcl-2、Mcl-1和Bax蛋白表达。结果与结论:(1)假手术组大鼠心肌梗死面积0%,与模型组和空白载体组比较,Adv-MRTF-A组大鼠心肌梗死面积明显减少(P<0.001);心肌细胞凋亡率显著降低(P<0.001);心肌组织中MRTF-A、Bcl-2、Mc-1蛋白相对表达量均升高,而Bax蛋白相对表达量降低(P<0.05)。(2)结果说明,重组腺病毒介导MRTF-A基因表达可有效减少心肌缺血再灌注损伤大鼠心肌梗死面积,其机制可能与促进Bcl-2、Mcl-1蛋白表达、抑制Bax蛋白表达从而抗心肌细胞凋亡有关。BACKGROUND： Up-regulation of m RNA expression of myocardin-related transcription factor-A（MRTF-A） has been shown to be inhibit neuronal apoptosis after cerebral ischemia/reperfusion, and play a protective role in the brain. However, its effect on the apoptosis of cardiomyocytes after ischemia/reperfusion injury remains unclear. OBJECTIVE： To investigate the effect of recombinant adenovirus encoding murine MRTF-A on the cardiomyocyte apoptosis induced by ischemia/reperfusion injury in rats. METHODS： Sprague-Dawley rats were randomly divided into sham operation, model, blank vector and Adv-MRTF-A groups. Recombinant adenovirus encoding MRTF-A was prepared, and the models of ischemia/reperfusion injury were constructed in the rats except the sham operation group. The virus titer 1.2×10^10 TU/mL of Adv-MRTF-A vector was injected into the infracted part on the left ventricular free wall in the Adv-MRTF-A group, while the rats in the blank vector group was given the injection of the same amount of blank virus. The myocardial infarct area was detected by TTC staining. The apoptosis of cardiomyocytes was detected by TUNEL method. The expression levels of MRTF-A, Bcl-2, Mcl-1 and Bax proteins in the myocardium were detected using western blot assay. RESULTS AND CONCLUSION： The myocardial infarct area in the sham operation group was 0%. Compared with the model and blank vector groups, the myocardial infarct area in the Adv-MRTF-A group was significantly decreased（P〈0.001）, the apoptotic rate of cardiomyocytes was significantly decreased（P〈0.001）; the relative expression levels of MRTF-A, Bcl-2 and Mcl-1 proteins in the in the myocardium were significantly up-regulated, while the relative expression level of Bax protein was significantly down-regulated（P〈0.05）. In summary, recombinant adenovirus encoding MRTF-A effectively reduces myocardial infarct ares in rats with myocardial ischemia/reperfusion injury, probably by promoting the expression levels of Bcl-2 and Mcl-1 proteins, and inhibiting

关 键 词：缺血再灌注损伤 MRTF-A 实验动物 大鼠 腺病毒载体 细胞凋亡 组织构建 

分 类 号：R318[医药卫生—生物医学工程]