lncRNA MAGI2-AS3靶向调控miR-130a-5p对柯萨奇病毒B3诱导的大鼠心肌细胞H9C2损伤的影响  

Effects of lncRNA MAGI2-AS3 targeting miR-130a-5p on coxsackie virus B3-induced rat cardiomyocyte H9C2 injury

作　　者：朱康宁[1] 郑培明 高岚[1] ZHU Kangning;ZHENG Peiming;GAO Lan(Department of Laboratory,Henan Provincial People′s Hospital,Zhengzhou 450003)

机构地区：[1]河南省人民医院检验科,郑州450003

出　　处：《郑州大学学报(医学版)》2023年第6期801-806,共6页Journal of Zhengzhou University(Medical Sciences)

基　　金：国家自然科学基金项目(81802094)。

摘　　要：目的:探讨lncRNA MAGI2-AS3靶向miR-130a-5p调控柯萨奇病毒B3(CVB3)诱导的大鼠心肌细胞H9C2损伤的分子机制。方法:对照组,用不含药物、病毒的DMEM处理H9C2细胞48 h。CVB3组,用含有100倍半数组织培养感染剂量CVB3的DMEM处理H9C2细胞48 h(感染)。CVB3+sh-NC组,H9C2细胞转染sh-NC慢病毒后感染CVB3。CVB3+sh-MAGI2-AS3组,H9C2细胞转染sh-MAGI2-AS3慢病毒后感染CVB3。CVB3+miR-NC组,H9C2细胞转染miR-NC后感染CVB3。CVB3+miR-130a-5p组,H9C2细胞转染miR-130a-5p mimic后感染CVB3。sh-MAGI2-AS3+抗miR-NC组,sh-MAGI2-AS3、抗miR-NC共转染H9C2细胞后感染CVB3。sh-MAGI2-AS3+抗miR-130a-5p组,sh-MAGI2-AS3、抗miR-130a-5p共转染H9C2细胞后感染CVB3。qRT-PCR检测MAGI2-AS3、miR-130a-5p相对表达量;Annexin V/PI双染法检测细胞凋亡率;ELISA法检测TNF-α、IL-6分泌水平;双荧光素酶报告实验验证MAGI2-AS3与miR-130a-5p靶向关系;Western blot检测Bcl-2、Cleaved-Caspase-3蛋白相对表达量。结果:与对照组比较,CVB3组MAGI2-AS3相对表达量、细胞凋亡率、Cleaved-Caspase-3蛋白表达水平及TNF-α、IL-6分泌水平升高,而miR-130a-5p相对表达量、Bcl-2蛋白表达水平降低(P<0.05)。分别与CVB3+sh-NC组、CVB3+miR-NC组相比,CVB3+sh-MAGI2-AS3组、CVB3+miR-130a-5p组细胞凋亡率、Cleaved-Caspase-3蛋白表达水平及TNF-α、IL-6分泌水平降低,Bcl-2蛋白表达水平升高(P<0.05)。MAGI2-AS3可靶向调控miR-130a-5p表达。与sh-MAGI2-AS3+抗miR-NC组比较,sh-MAGI2-AS3+抗miR-130a-5p组细胞凋亡率、Cleaved-Caspase-3蛋白表达水平及TNF-α、IL-6分泌水平升高,Bcl-2蛋白表达水平降低(P<0.05)。结论:沉默MAGI2-AS3可通过上调miR-130a-5p而抑制细胞凋亡、炎症反应,进而减轻CVB3诱导的H9C2细胞损伤。Aim:To explore the molecular mechanism of lncRNA MAGI2-AS3 targeting miR-130a-5p to regulate coxsackie virus B3(CVB3)-induced rat cardiomyocyte H9C2 injury.Methods:H9C2 cells were allocated into different groups and treated for 48 hours.Control group,DMEM treatment without drugs and viruses.CVB3 group,DMEM treatment with 100-fold 50% tissue culture infective dose of CVB3.CVB3+sh-NC group,treatment with CVB3 after sh-NC transfection.CVB3+sh-MAGI2-AS3 group,treatment with CVB3 after sh-MAGI2-AS3 transfection.CVB3+miR-NC group,treatment with CVB3 after miR-NC transfection.CVB3+miR-130a-5p group,CVB3 treatment after miR-130a-5p mimic transfection.sh-MAGI2-AS3+anti-miR-NC group,CVB3 treatment after sh-MAGI2-AS3,anti-miR-NC co-transfection.sh-MAGI2-AS3+anti-miR-130a-5p group,CVB3 treatment after sh-MAGI2-AS3,anti-miR-130a-5p co-transfection.MAGI2-AS3 and miR-130a-5p expression were detected by qRT-PCR.Annexin V/PI staining was used to detect the rate of apoptosis.The levels of TNF-α and IL-6 secreted were detected by ELISA.A dual-luciferase reporter assay was used to detect the targeting relationship between MAGI2-AS3 and miR-130a-5p.The protein expressions of Bcl-2 and Cleaved-Caspase-3 were detected by Western blot.Results:Compared with the control group,the expression of MAGI2-AS3,the rate of apoptosis,the protein level of Cleaved-Caspase-3,the levels of TNF-α and IL-6 secreted in the CVB3 group increased,while the expression of miR-130a-5p,the protein level of Bcl-2 decreased(P<0.05).Compared with CVB3+sh-NC group and CVB3+miR-NC group,the rate of apoptosis,the protein level of Cleaved-Caspase-3,and the levels of TNF-α and IL-6 secreted in the CVB3+sh-MAGI2-AS3 group and CVB3+miR-130a-5p group decreased,while the protein level of Bcl-2 increased(P<0.05).MAGI2-AS3 could target and regulate the expression of miR-130a-5p.Compared with sh-MAGI2-AS3+anti-miR-NC group,the rate of apoptosis,the protein level of Cleaved-Caspase-3,the levels of TNF-α and IL-6 secreted in the sh-MAGI2-AS3+anti-miR-130a-5p group increased

关 键 词：MAGI2-AS3 miR-130a-5p 柯萨奇病毒B3 大鼠 心肌细胞 

分 类 号：R373.23[医药卫生—病原生物学]