IDI2-AS1通过微小RNA-33b-5p调控NR4A2影响急性心肌梗死的发展  

IDI2-AS1 influences the development of acute myocardial infarction by regulating NR4A2 through microRNA-33b-5p

作　　者：吴树兴 庞志华[1] 王茹[1] 崔健 黎文婷[1] 杨霄羽 姚朱华[1] Wu Shuxing;Pang Zhihua;Wang Ru;Cui Jian;Li Wenting;Yang Xiaoyu;Yao Zhuhua(Department of Cardiology 1,Tianjin People's Hospital,Tianjin 300131,China)

机构地区：[1]天津市人民医院心脏内科1,天津300131

出　　处：《中华危重病急救医学》2024年第9期972-979,共8页Chinese Critical Care Medicine

基　　金：天津市科技计划项目(22JCYBJC00580)。

摘　　要：目的探究长链非编码RNA(lncRNA)IDI2-AS1/微小RNA-33b-5p(miR-33b-5p)/核受体相关蛋白NR4A2竞争性内源RNA(ceRNA)调控网络对急性心肌梗死(AMI)的影响及相关性,验证IDI2-AS1通过miR-33b-5p调控NR4A2影响心肌梗死的发生发展。方法通过基因表达数据库(GEO)获取心肌梗死相关的miRNA及mRNA表达芯片,并进行差异表达分析;通过TargetScan数据库对NR4A2的上游调控机制进行预测。按随机数字表法将32只C57/BL6雄性小鼠分为假手术(Sham)组、AMI模型组、miR-33b-5p mimic组(结扎过程中心脏组织局部注射miR-33b-5p mimic慢病毒5×10^(7) TU)和miR-33b-5p inhibitor组(结扎过程中心脏组织局部注射miR-33b-5p inhibitor慢病毒5×10^(7) TU),每组8只。超声心动图观察各组小鼠心脏左室舒张期末内径(LVEDD)和左室收缩期末内径(LVESD),计算左室短轴缩短率(LVFS)和左室射血分数(LVEF)。末次称重后麻醉处死小鼠并取心脏组织,光镜下观察心脏组织Masson染色情况,计算心肌胶原容积分数(CVF)和心肌梗死面积。收集SPF级SD大鼠乳鼠心肌细胞,分为正常对照组(Control组)、缺血缺氧模型组、miR-33b-5p mimic转染组(缺血缺氧处理前转染miR-33b-5p mimic)和miR-33b-5p inhibitor转染组(缺血缺氧处理前转染miR-33b-5p inhibitor),测定心肌细胞天冬氨酸特异性半胱氨酸蛋白酶3/7(caspase-3/7)活性。采用酶联免疫吸附试验(ELISA)检测白细胞介素(IL-1β、IL-6)和肿瘤坏死因子-α(TNF-α)水平;采用比色法检测丙二醛(MDA)、超氧化物歧化酶(SOD)、肌酸激酶(CK)、肌酸激酶同工酶(CK-MB)、乳酸脱氢酶(LDH)水平;采用实时荧光定量聚合酶链反应(RT-qPCR)检测凋亡相关蛋白Bax和Bcl-2、细胞色素C(Cyt C)及IDI2-AS1/miR-33b-5p/NR4A2调控轴基因表达。结果心肌梗死芯片分析显示,NR4A2在心肌梗死中表达显著上调,上游调控机制预测表明其可能通过IDI2-AS1/miR-33b-5p/NR4A2调控轴影响心肌梗死的发生发展。超声心动图�Objective To explore the effect and correlation of long non-coding RNA(lncRNA)IDI2-AS1/microRNA-33b-5p(miR-33b-5p)/nuclear receptor-associated protein NR4A2 competitive endogenous RNA(ceRNA)regulatory network on acute myocardial infarction(AMI),and to verify whether IDI2-AS1 regulates NR4A2 through miR-33b-5p to affect the occurrence and development of myocardial infarction.Methods The miRNA and mRNA expression chips related to myocardial infarction were obtained from gene expression omnibus(GEO),and the differential expression was analyzed.The upstream regulatory mechanism of NR4A2 was predicted using TargetScan database.Thirty-two male C57/BL6 mice were divided into Sham group,AMI model group,miR-33b-5p mimic group[miR-33b-5p mimic lentivirus(5×10^(7) TU)was injected locally into the heart tissue during ligation]and miR-33b-5p inhibitor group[miR-33b-5p inhibitor lentivirus(5×10^(7) TU)was injected locally into the heart tissue during ligation]according to random number table method,with 8 mice per group.Left ventricular end-diastolic diameter(LVEDD)and left ventricular end-systolic diameter(LVESD)were asseessed by echocardiography,left ventricular fractional shortening(LVFS)and left ventricular ejection fraction(LVEF)were calculated.After the last weighing,the anesthetized mice were sacrificed and the heart tissues were taken.Masson staining of the heart tissues was observed under light microscope,myocardial collagen volume fraction(CVF)and infarct size were calculated.Cardiomyocytes of SPF grade SD rats were collected.They were divided into normal control group(control group),ischemia-hypoxia model group,miR-33b-5p mimic transfection group(miR-33b-5p mimic transfection group before ischemia and hypoxia treatment)and miR-33b-5p inhibitor transfection group(miR-33b-5p inhibitor transfection group before ischemia and hypoxia treatment).The activity of caspase-3/7 in cardiomyocytes was measured.The levels of interleukins(IL-1β,IL-6)and tumor necrosis factor-α(TNF-α)were detected by enzyme-linked immunosorben

关 键 词：IDI2-AS1/微小RNA-33b-5p/NR4A2内源竞争RNA调控网络 急性心肌梗死 小鼠模型 细胞缺氧模型 

分 类 号：R542.22[医药卫生—心血管疾病]