miR-30e-5p靶向BIM调控缺氧诱导细胞凋亡的应用研究  

Application Study of miR-30e-5p Targeted BIM in Regulating Hypoxia-Induced Apoptosis

作　　者：吴宁夏 李飞[2] 

机构地区：[1]延安大学医学院,陕西 延安 [2]延安大学附属医院心血管内科,陕西 延安

出　　处：《临床医学进展》2024年第6期344-352,共9页Advances in Clinical Medicine

摘　　要：目的:研究miR-30e-5p与H2O2损伤后心肌细胞凋亡的关系,探讨miR-30e-5p靶向BIM基因调控H2O2损伤后心肌细胞凋亡的作用,为心肌梗死的诊断以及治疗提供新的靶点。方法:将40天CA16分为正常组(N)、缺氧组(H)、过表达对照组(NC+H)、miR-30e-5p过表达组(miR+H)。N组置于正常培养箱下培养;H组置于缺氧培养箱培养24 h;NC+H组与miR+H组分别为稳定感染装载有miR-NC和miR-30e-5p的重组慢病毒后缺氧处理24 h。应用RT-qPCR检测CA16中miR-30e-5p和BIM的mRNA表达水平,应用Caspase-3活性实验和流式细胞技术检测hiPSC-CMs的凋亡水平。利用生物信息学miRNA数据库预测miR-30e-5p的靶基因,并应用双荧光素酶报告基因实验进行验证。结果:1) 与N组比较,H组miR-30e-5p表达水平明显下调;与NC+H组比较,miR+H组miR-30e-5p表达水平则明显上调。2) 与N组比较,H组Caspase-3活性水平明显升高;与NC+H组比较,miR+H组Caspase-3活性水平下降。3) 与N组比较,H组CA16细胞凋亡比例明显上升;与NC+H组比较,miR+H组CA16凋亡比例则明显下降。4) 与N组比较,H组蛋白BIM表达水平上调;与NC+H组比较,miR+H组蛋白BIM表达水平下调。5) 通过miRNA数据库分析发现BIM是miR-30e-5p的潜在靶基因之一。结论:1) miR-30e-5p调控缺氧诱导CA16的凋亡。2) BIM是miR-30e-5p的靶基因之一。3) miR-30e-5p靶向BIM调控缺氧诱导CA16的凋亡。4) miR-30e-5p在急性心肌梗死的发生、发展过程中可能发挥抑癌基因的作用。Objective: To investigate the relationship between miR-30e-5p and myocardial cell apoptosis after H2O2 injury, and to explore the role of miR-30e-5p targeting BIM genes in regulating myocardial cell apoptosis after H2O2 injury, providing new targets for the diagnosis and treatment of myocardial infarction. Method: The 40 day CA16 was divided into normal group (N), hypoxia group (H), overexpression control group (NC+H), and miR-30e-5p overexpression group (miR+H). Group N was cultured in a normal incubator;Group H was incubated in a hypoxia incubator for 24 hours;The NC+H group and miR+H group were treated with hypoxia for 24 hours after stable infection with recombinant lentivirus loaded with miR-NC and miR-30e-5p, respectively. RT qPCR was used to detect the mRNA expression levels of miR-30e-5p and BIM in CA16, and Caspase-3 activity assay and flow cytometry were used to detect the apoptosis level of hiPSC-CMs. Using bioinformatics miRNA database to predict target genes of miR-30e-5p, and validating with dual luciferase reporter gene experiments. Result: 1) Compared with group N, the expression level of miR-30e-5p in group H was significantly down-regulated;Compared with the NC+H group, the expression level of miR-30E-5p in the miR+H group was significantly up-regulated. 2) Compared with group N, the activity level of Caspase-3 in group H was significantly increased;Compared with NC+H group, the activity level of Caspase-3 in miR+H group was decreased. 3) Compared with group N, the apoptosis ratio of CA16 cells in group H was significantly increased;Compared with NC+H group, the apoptosis ratio of CA16 in miR+H group was significantly decreased. 4) Compared with group N, BIM expression level of histone H was up-regulated;Compared with NC+H group, BIM expression level of histone in miR+H was down-regulated. 5) The analysis of miRNA database showed that BIM was one of the potential target genes of miR-30e-5p. Conclusion: 1) miR-30e-5p regulates apoptosis of CA16 induced by hypoxia. 2) BIM is one of the target gene

关 键 词：miR-30e-5p CA16 抑制缺氧 凋亡 

分 类 号：R73[医药卫生—肿瘤]