miR-92a在心肌缺血再灌注损伤中作用及机制研究  

The role and mechanism of miR-92 a in myocardial ischemia-reperfusion injury

作　　者：周丽 王诗奇 邹武松 Zhou Li;Wang Shi-qi;Zou Wu-song(Department of Internal Medicine,Pu'ai Hospital,Tongji Medical College,Huazhong University of Science and Technology,Wuhan 4th Hospital,Wuhan 430033,China)

机构地区：[1]武汉市第四医院华中科技大学同济医学院附属普爱医院内科,湖北武汉430033

出　　处：《中国急救医学》2020年第3期254-259,共6页Chinese Journal of Critical Care Medicine

基　　金：武汉市卫生和计划生育委员会科研项目(WX18D49)。

摘　　要：目的探究miR-92a在心肌缺血再灌注损伤(MIRI)中作用及机制.方法60只小鼠随机分为4组,分别为对照组(假手术组)、模型组、模型+阴性对照(negative control,NC)组和模型+miR-92a抑制物(inhibitor)组.通过结扎冠脉降支的方法建立小鼠MIRI模型.模型+NC组和模型+inhibitor组在建模前12 h静脉注射空载质粒和miR-92a inhibitor质粒.检测建模后24 h每组小鼠的心脏功能、血清心肌酶水平、心肌梗死面积.使用流式细胞术检测细胞凋亡情况.使用RT-qPCR和Western blot检测JAK-STAT通路和炎症因子mRNA和蛋白水平.结果建模后小鼠心脏射血分数[EF,(43.35±2.98)%]和缩短分数[FS,(21.23±2.11)%]均显著下降(P<0.05),梗死百分比为(40.05±3.65)%.模型+inhibitor组EF(62.64±4.38)%和FS(32.45±2.29)%显著高于模型组和模型+NC组(P<0.05),梗死百分比(18.66+3.11)%显著低于模型组和模型+NC组(P<0.05).模型组肌酸激酶[CK,(0.51±0.07)U/mL]和乳酸脱氢酶[LDH,(517.76±35.37)U/L]显著高于对照组(P<0.05).模型+inhibitor组血清CK(0.39±0.05)U/mL和LDH(457.67±35.29)U/L显著低于模型组和模型+NC组(P<0.05).建模24 h后小鼠心肌细胞中miR-92a水平显著升高(P<0.05),而模型+inhibitor组miR-92a水平(2.94±0.78)显著低于模型组和模型+NC组(P<0.05).建模后小鼠心肌细胞凋亡率(31.83±2.76)%显著升高(P<0.05),模型+inhibitor组心肌细胞凋亡率(18.88±1.84)%显著低于模型组和模型+NC组(P<0.05).模型+inhibitor组TNF-α(1.33±0.19)和IL-6(1.44±0.19)和mRNA(1.25±0.18,1.23±0.15)显著低于模型组和模型+NC组(P<0.05).网站预测和荧光素酶报告验证了miR-92a靶向PTEN.建模后小鼠心肌组织中PTEN水平降低而JAK和STAT蛋白磷酸化水平显著增加(P<0.05),模型+inhibitor组PTEN水平升高而p-JAK/JAK和p-STAT/STAT水平显著低于模型组和模型+NC组(P<0.05).结论抑制miR-92a具有保护心肌细胞减少MIRI的作用,这可能是由于抑制miR-92a可通过上调PTEN的表达抑制JAK2-STAT3通路的活Objective To investigate the role and mechanism of miR-92a in myocardial ischem ia-reperfusion injury(MIRI).Methods Sixty mice were randomly divided into 4 groups:control group(sham operation group),model group,model+negative control(NC)group and model+miR-92 a inhibitor(inhibitor)group.MIRI models were established in mice by ligation of coronary descending.The model+NC group and the model+inhibitor group were injected with the empty plasmid and the miR-92 a inhibitor plasmid at 12 h before the modeling.Cardiac function,serum myocardial enzyme levels,and myocardial infarct size were measured in mice of each group after modeling for 24 hours.Apoptosis was detected by flow cytometry.The levels of JAK-STAT pathway and inflammatory factor mRNA and protein were detected by RT-qPCR and Western blot.Results After m odeling,the cardiac ejection fraction[E F,(43.35±2.98)%]and shortened fraction[F S,(21.23±2.11)%of the mice were significantly decreased(P<0.05),and the infarct percentage(40.05±3.65)%was increased.The EF(62.64±4.38)%and FS(32.45±2.29)%of the model+inhibitor group were significantly higher than those of the model group and model+NC group(P<0.05),but the infarct percentage(18.66±3.11)%was decreased(P<0.05).Creatine kinase[C K,(0.51±0.07)U/m L]and lactate dehydrogenase[LDH,(517.76±35.37)U/L]in the model group were significantly higher than those in the control group(P<0.05).The serum CK(0.39±0.05)U/m L and LDH(457.67±35.29)U/L in the model+inhibitor group were significantly lower than those in the model group and the model+NC group(P<0.05).The level of miR-92a in mouse cardiomyocytes increased significantly after modeling for 24 h(P<0.05).The level of miR-92a in the model+inhibitor group(2.94±0.78)was significantly lower than that in the model group and the model+NC group(P<0.05).Apoptosis rate of mouse cardiomyocytes(31.83±2.76)%increased significantly after modeling(P<0.05).The cardiomyocyte apoptosis rate(18.88±1.84)%in the model+inhibitor group was significantly lower than that in the model gro

关 键 词：miR-92a 心肌缺血 心肌缺血再灌注损伤(MIRI) JAK-STAT 

分 类 号：R542.2[医药卫生—心血管疾病]