miR-133a-3p修饰的间充质干细胞外泌体促进心肌梗死后大鼠心肌修复  

Exosomes derived from miR-133a-3p engineered mesenchymal stem cells promote myocardial repair in rats after acute myocardial infarction

作　　者：孙铃 朱文武 张健 赵鹏程 朱叶倩 张凤祥 Ling Sun;Wenwu Zhu;Jian Zhang;Pengcheng Zhao;Yeqian Zhu;Fengxiang Zhang(Department of Cardiology,the First Affiliated Hospital of Nanjing Medical University,Nanjing 210029,China;Department of Cardiology,Xuzhou Central Hospital,Xuzhou Clinical School of Nanjing Medical University,Xuzhou 221000,China)

机构地区：[1]南京医科大学第一附属医院、江苏省人民医院心血管内科,南京210029 [2]徐州市中心医院心血管内科,徐州221000

出　　处：《中华心血管病杂志》2024年第1期72-78,共7页Chinese Journal of Cardiology

基　　金：国家自然科学基金(81901410,81871113);江苏省自然基金(BK20201489)。

摘　　要：目的探讨miR-133a-3p修饰的人源脐带血间充质干细胞(ucMSC)衍生的外泌体对急性心肌梗死(AMI)后大鼠心肌修复的作用。方法体外扩增培养ucMSC,将携带miR-133a-3p的慢病毒载体及阴性对照载体分别转染至ucMSC,分别提取其分泌的外泌体,命名为miR-133a-3p-Exo及miR-NC-Exo。采用冠状动脉左前降支结扎术构建大鼠AMI模型,分别于梗死区边缘原位注射miR-133a-3p-Exo或miR-NC-Exo进行干预。干预后28 d,采用超声心动图检测大鼠心功能;采用Masson染色检测大鼠梗死后心肌纤维化面积;采用TUNEL染色评估梗死后大鼠心肌凋亡情况;采用免疫荧光染色评估梗死后大鼠血管新生情况。结果与miR-NC-Exo组相比,miR-133a-3p-Exo组在AMI后28 d左心室射血分数较高[(64.2%±8.9%)比(47.4%±9.8%),P<0.05],心肌纤维化面积[(18.0%±1.5%)比(31.2%±7.3%),P<0.01]和凋亡率[(15.1%±4.4%)比(25.6%±3.6%),P<0.05]较低;梗死边缘区血小板-内皮黏附因子(CD31)及平滑肌肌动蛋白(α-SMA)阳性血管的数量均较多[CD31:(13.9±2.0)条比(2.9±0.9)条,α-SMA:(11.0±1.6)条比(3.5±0.9)条,P均<0.0001]。结论miR-133a-3p修饰的ucMSC分泌的外泌体可有效抑制梗死后心肌凋亡、促进梗死后血管新生,从而改善AMI后大鼠的心功能。Objective To investigate the effects of exosome derived from miR-133a-3p engineered human umbilical cord blood mesenchymal stem cells(ucMSC)on myocardial repair after acute myocardial infarction(AMI)in rats.Methods UcMSC was amplified and cultured in vitro.Lentiviral carrying miR-133a-3p and negative control vectors were transfected into ucMSC.Exosomes secreted by the transfected ucMSC were named miR-133a-3p-Exo and miR-NC-Exo,respectively.The AMI model of rats was established by ligation of the left anterior descending coronary artery.MiR-133a-3p-Exo or miR-NC-Exo were then injected into the border zone of the infarct area.Cardiac function was assessed by echocardiography after twenty-eight days of intervention,and Masson staining was used to evaluate the area of myocardial fibrosis post-AMI.The myocardial apoptosis after infarction was evaluated by TUNEL staining and the angiogenesis after infarction was evaluated by immunofluorescence staining in the current study.Results Compared with the miR-NC-Exo group,the left ventricular ejection fraction in the miR-133a-3p-Exo group was significantly increased((47.4%±9.8%)vs.(64.2%±8.9%),P<0.05).While the myocardial fibrosis area((31.2%±7.3%)vs.(18.0%±1.5%),P<0.01)and the percentage of apoptotic cardiomyocytes((25.6%±3.6%)vs.(15.1%±4.4%),P<0.05)was significantly reduced in the miR-133a-Exo group.Besides,the expression of CD31 andα-smooth muscle actin(α-SMA)were also increased significantly in the miR-133a-3p-Exo group compared to the miR-NC-Exo group(CD31:(2.9±0.9)vs.(13.9±2.0),P<0.0001,α-SMA:(3.5±0.9)vs.(11.0±1.6),P<0.0001).Conclusion Exosome derived from miR-133a-3p engineered ucMSC effectively inhibited myocardial apoptosis and promoted angiogenesis,thus improving the cardiac function after myocardial infarction in rats.

关 键 词：微RNAS 间充质干细胞 外泌体 急性心肌梗死 心肌修复 

分 类 号：R542.22[医药卫生—心血管疾病]