circRNA001653对急性心肌梗死心肌细胞凋亡的作用及机制  

Effects of circRNA_001653 on myocardial cell apoptosis in acute myocardial infarction and its mechanism

作　　者：李川 谢景臣 赵展庆 LI Chuan;XIE Jingchen;ZHAO Zhanqing(Intensive Care Unit,Hainan West Central Hospital,Danzhou 571799,Hainan,China;Department of Respiratory,Hainan West Central Hospital,Danzhou 571799,Hainan,China)

机构地区：[1]海南西部中心医院重症医学科,海南儋州571799 [2]海南西部中心医院呼吸内科,海南儋州571799

出　　处：《贵州医科大学学报》2023年第5期527-536,共10页Journal of Guizhou Medical University

基　　金：海南省自然科学基金面上项目(820MS154)。

摘　　要：目的探讨环状RNA(circRNA)_001653(circ_001653)对急性心肌梗死(AMI)心肌细胞凋亡的作用及机制。方法采用生物信息学分析AMI时差异表达的circRNA;大鼠心肌细胞分别培养于常氧(Normoxia组)和缺氧12 h(Hypoxia 12 h组)、24 h(Hypoxia 24 h组)、48 h(Hypoxia 48 h组)条件下,实时荧光定量PCR(RT-qPCR)检测上述4组细胞中circ_001653、微小RNA(microRNA)-324-5p、ELAV样RNA结合蛋白1(ELAVL1)的表达;选取缺氧48 h的心肌细胞转染sh-NC(sh-NC组)、sh-circ_001653(sh-circ_001653组)、circ-NC(circ-NC组)、circ_001653(circ_001653组)、miR-NC(miR-NC组)、miR-324-5p mimic(miR-324-5p mimic组)、circ_001653+miR-NC(circ_001653+miR-NC组)、circ_001653+miR-324-5p mimic(circ_001653+miR-324-5p mimic组)、miR-324-5p mimic+pcDNA3.1(miR-324-5p mimic+pcDNA3.1组)及miR-324-5p mimic+ELAVL1(miR-324-5p mimic+ELAVL1组)等载体,采用比色试剂盒和流式细胞术分别检测Normoxia组、Hypoxia-48 h组、sh-NC组、sh-circ_001653组、circ-NC组、circ_001653组、miR-NC组、miR-324-5p mimic组、circ_001653+miR-NC组、circ_001653+miR-324-5p mimic组、miR-324-5p mimic+pcDNA3.1组、miR-324-5p mimic+ELAVL1组心肌细胞中天冬氨酸特异性半胱氨酸蛋白酶-3(Caspase-3)、Caspase-9活性及细胞凋亡水平。结果生物信息学分析结果显示,与正常组比较、circ_001653在AMI中表达增强(P<0.05);与Normoxia组比较,Hypoxia 48 h组细胞中circ_001653和ELAVL1表达升高、miR-324-5p表达降低(P<0.05);与Normoxia组比较,Hypoxia 48 h组细胞凋亡率与Caspase-3、Caspase-9活力均升高(P<0.05);与sh-NC组比较,sh-circ_001653组细胞凋亡率和Caspase-3、Caspase-9活力均降低(P<0.05);与circ-NC组比较,circ_001653组细胞凋亡率和Caspase-3、Caspase-9活力均升高(P<0.05);与circ_001653+miR-NC组比较,circ_001653+miR-324-5p mimic组细胞凋亡率和Caspase-3、Caspase-9活力均降低(P<0.05);与miR-NC组比较,miR-324-5p mimic组细胞凋亡率和Caspase-3、Caspase-9活力均降低(P<0.05);与miR-324-5p mimic+pcDNObjective To explore the effects and mechanism of circular RNA(circRNA)_001653(circ_001653)on myocardial cell apoptosis in acute myocardial infarction(AMI).Methods The differentially expressed circRNAs in AMI were analyzed by bioinformatic tools.Rat cardiomyocytes were purchased and cultured under the conditions of normal oxygen(Normoxia group)and Hypoxia 12 h(Hypoxia 12 h group),24 h(Hypoxia 24 h group)and 48 h(Hypoxia 48 h group),respectively.The expression of circ_001653,microRNA-324-5p,and ElAV-like RNA binding protein 1(ELAVL1)in the above four groups was detected by real-time quantitative PCR(RT-qPCR).Cardiomyocytes were selected after 48 h of hypoxia and transfected with the following vector sh-NC(sh-NC group),sh-circ_001653(sh-circ_001653 group),circ-NC(circ-NC group),circ_001653(circ_001653group),miR-NC(miR-NC group),miR-324-5p mimic(miR-324-5p mimic group),circ_001653+miR-NC(circ_001653+miR-NC group),circ_001653+miR-324-5p mimic(circ_001653+miR-324-5p mimic group),miR-324-5p mimic+pcDNA3.1(miR-324-5p mimic+pcDNA3.1 group),and miR-324-5p mimic+ELAVL1(miR-324-5p mimic+ELAVL1 group).Colorimetric kit and flow cytometry were used to detect the aspartic acid-specific cysteine proteinase-3(Caspase-3),Caspase-9 activity and apoptosis level respectively in Normoxia group,Hypoxia-48 h group,sh-NC group,sh-circ_001653 group,circ-NC group,circ-001653 group,miR-NC group,miR-324-5p mimic group,circ_001653+miR-NC group,circ_001653+miR-324-5p mimic group,miR-324-5p mimic+pcDNA3.1 group,and miR-324-5p mimic+ELAVL1 group.Results Bioinformatics analysis showed that the expression of circ-001653 was enhanced in AMI compared with the normal group(P<0.05).Compared with Normoxia group,the expression of circ_001653 and ELAVL1 increased,but the expression of miR-324-5p decreased in Hypoxia 48 h group(all P<0.05).Compared with Normoxia group,the apoptosis rate and Caspase-3 and Caspase-9 activities in Hypoxia 48 h group increased(all P<0.05).Compared with sh-NC group,the apoptosis rate and the activities of Caspase-3 and Caspase

关 键 词：细胞凋亡 急性心肌梗死 心肌细胞 缺氧 环状RNA_001653 ELAV RNA结合蛋白1 

分 类 号：R542.2[医药卫生—心血管疾病]