miR-30a-5p靶向KDM3A缓解缺氧诱导H9c2细胞凋亡  

miR-30a-5p targets KDM3A to alleviate hypoxia-induced apoptosis in H9c2 cells

作　　者：罗梦圆 张辛颖 张彬 邵明学[1] 田乃亮[1] LUO Mengyuan;ZHANG Xinying;ZHANG Bin;SHAO Mingxue;TIAN Nailiang(Department of Cardiology,Nanjing First Hospital,Nanjing Medical University,Nanjing 210006,Jiangsu,China;Department of Emergency Medicine,Nanjing First Hospital,Nanjing Medical University,Nanjing 210006,Jiangsu,China;Department of Cardiology,Nanjing Yuhua Hospital,Nanjing 210039,Jiangsu,China)

机构地区：[1]南京医科大学附属南京医院心血管内科,南京210006 [2]南京医科大学附属南京医院急诊医学科,南京210006 [3]南京市雨花医院心内科,南京210039

出　　处：《中国分子心脏病学杂志》2025年第1期6637-6645,共9页Molecular Cardiology of China

基　　金：南京市卫生科技发展专项资金项目(YKK21130)。

摘　　要：目的探讨miR-30a-5p通过靶向KDM3A调节心肌细胞在低氧条件下的凋亡机制。方法构建急性心肌梗死(acute myocardial infarction,AMI)小鼠模型并模拟缺氧环境于小鼠心肌原代细胞中,采用qPCR技术定量分析miR-30a-5p的表达水平。同时,结合生物信息学分析预测miR-30a-5p与KDM3A 3′UTR的结合位点,并通过双荧光素酶报告实验验证二者的结合关系。在H9c2心肌细胞株中模拟低氧环境,建立缺氧诱导的细胞凋亡模型,采用qPCR分析miR-30a-5p的表达变化,并通过Western blot检测凋亡相关蛋白(Bax、Bcl-2、c-PARP1)的表达水平。利用TUNEL染色法评估低氧诱导的细胞凋亡程度。低氧处理前,分别转染H9c2细胞miR-30a-5p模拟物/抑制剂和KDM3A siRNA,以评估其对心肌细胞凋亡的影响。结果在小鼠AMI模型、缺氧诱导的小鼠心肌原代细胞及H9c2心肌细胞中,miR-30a-5p的表达显著下降。双荧光素酶实验验证了miR-30a-5p可与KDM3A 3′UTR结合。进一步分析显示,在低氧条件下,KDM3A蛋白的表达水平显著升高。Bax/Bcl-2比值的增加以及c-PARP1蛋白的升高与细胞凋亡率的增加密切相关。过表达miR-30a-5p能够显著逆转低氧诱导的心肌细胞凋亡相关蛋白的变化,而抑制miR-30a-5p则加剧了这些效应。敲低KDM3A亦能减轻低氧诱导的心肌细胞凋亡。结论miR-30a-5p通过靶向KDM3A调控心肌细胞在低氧条件下的凋亡过程,为AMI的治疗提供了新的分子靶点。Objective To explore whether miR-30a-5p regulates cardiomyocyte apoptosis under hypoxic conditions by targeting KDM3A.Methods Acute myocardial infarction(MI)mouse models were established,and hypoxic conditions were simulated in primary mouse cardiomyocytes.qPCR was used to quantify the expression level of miR-30a-5p.Bioinformatics analysis predicted the binding site of miR-30a-5p in the 3′UTR of KDM3A,and dual-luciferase reporter assays were performed to confirm their interaction.In H9c2 cardiomyocyte cells,hypoxia was induced to establish a hypoxia-induced apoptosis model.qPCR was employed to assess the expression of miR-30a-5p,and Western blotting was used to detect the apoptosis-related proteins Bax,Bcl-2,and c-PARP1.TUNEL staining was performed to evaluate the degree of hypoxia-induced apoptosis.Prior to hypoxic treatment,H9c2 cells were transfected with miR-30a-5p mimics/inhibitors and KDM3A siRNA to assess their effects on cardiomyocyte apoptosis.Results The expression of miR-30a-5p was significantly decreased in the acute MI mouse model,hypoxia-induced primary mouse cardiomyocytes,and H9c2 cardiomyocyte cells.Dual-luciferase reporter assays confirmed that miR-30a-5p binds to the 3′UTR of KDM3A.Further analysis revealed that under hypoxic conditions,the expression of KDM3A protein was significantly increased.The increased Bax/Bcl-2 ratio and elevated c-PARP1 protein levels were closely associated with the increased apoptosis rate.Overexpression of miR-30a-5p significantly reversed the changes in apoptosis-related proteins induced by hypoxia,whereas inhibition of miR-30a-5p exacerbated these effects.Knockdown of KDM3A also alleviated hypoxia-induced cardiomyocyte apoptosis.Conclusions This study reveals thatmiR-30a-5p regulates cardiomyocyte apoptosis under hypoxic conditions by targeting KDM3A,providing a potential new moleculartarget for the treatment of myocardial infarction.

关 键 词：急性心肌梗死 缺氧 miR-30a-5p KDM3A 细胞凋亡 

分 类 号：R541.4[医药卫生—心血管疾病]