过表达miR-486-5p的骨髓间充质干细胞通过外泌体来保护缺氧损伤心肌细胞  

Bone Marrow Mesenchymal Stem Cell Overexpressing miR-486-5p Protect Hypoxic-Injured Cardiomyocytes via Exosomes

作　　者：姚思雨 张婷婷[2] 安松涛[1] YAO Siyu;ZHANG Tingting;AN Songtao(Central China Fuwai Hospital of Zhengzhou University,Zhengzhou 450003,Henan,China;Center for Clinical Single-Cell Biomedicine,Henan Provincial People’s Hospital,Zhengzhou 450003,Henan,China)

机构地区：[1]郑州大学华中阜外医院,河南郑州450003 [2]河南省人民医院临床单细胞生物医学中心,河南郑州450003

出　　处：《心血管病学进展》2024年第9期858-864,共7页Advances in Cardiovascular Diseases

基　　金：国家自然科学基金(81970312)。

摘　　要：目的探讨过表达miR-486-5p小鼠骨髓间充质干细胞(BMSC)来源外泌体(Exo)对缺氧损伤心肌细胞的影响及机制。方法通过慢病毒转染来构建过表达miR-486-5p的小鼠BMSC。提取转染成功的BMSC所分泌的Exo,包括空载体BMSC分泌的Exo(Exo-NC)、miR-486-5p过表达BMSC分泌的Exo(Exo-miR-486)及未处理BMSC分泌的Exo(Exo-Control)。取12~18 d胎鼠原代心肌细胞,将其分为常氧培养组(Normoxia组)、缺氧培养组(Hypoxia组)、Hypoxia+Exo-NC组及Hypoxia+Exo-miR-486组,并给予不同时间的缺氧处理来模拟心肌细胞缺氧状态,检测细胞增殖状态、凋亡率,以及活性氧(ROS)、核苷酸结合结构域富含亮氨酸重复序列和含热蛋白结构域受体3(NLRP3)和裂解的胱天蛋白酶-1(cleaved caspase-1)的表达水平。结果Hypoxia组心肌细胞增殖能力低于Normoxia组,而凋亡率及ROS、NLRP3和cleaved caspase-1的表达水平均高于Normoxia组(P<0.05);Hypoxia+Exo-NC组心肌细胞增殖能力高于Hypoxia组,凋亡率及ROS、NLRP3和cleaved caspase-1的表达水平低于Hypoxia组(P<0.05);Hypoxia+Exo-miR-486组心肌细胞增殖能力高于Hypoxia+Exo-NC组,凋亡率及ROS、NLRP3和cleaved caspase-1表达水平低于Hypoxia+Exo-NC组(P<0.05)。结论来自miR-486-5p过表达BMSC的Exo能减少缺氧导致的ROS的产生及其引起的心肌细胞凋亡,并改善缺氧引起的心肌细胞增殖缓慢的问题。Objective To investigate the effects and mechanisms of exosomes(Exo)derived from bone marrow mesenchymal stem cell(BMSC)in mice overexpressing miR-486-5p on hypoxic-injured cardiomyocytes.Methods Mouse BMSC overexpressing miR-486-5p was constructed by lentivirus transfection.Exo secreted by successfully transfected BMSC were extracted,including Exo secreted by empty carrier BMSC(Exo-NC),Exo secreted by BMSC overexpressing miR-486-5p(Exo-miR-486),and Exo secreted by untreated BMSC(Exo-Control).Primary cardiomyocytes from 12~18 d fetal rats were divided into normal oxygen culture group(Normoxia group),hypoxia culture group(Hypoxia group),Hypoxia+Exo-NC group and Hypoxia+Exo-miR-486 group,and were given hypoxia treatments for different times to stimulate the hypoxic state of cardiomyocytes.Cell proliferation,apoptosis rate,and the expression levels of reactive oxygen species(ROS),nucleotide-binding domain leucine-rich repeat and pyrin domain-containing receptor 3(NLRP3),and cleaved caspase-1 were examined.Results The proliferation of cardiomyocytes in Hypoxia group was lower than that in Normoxia group,while the apoptosis rate and the expression levels of ROS,NLRP3 and cleaved caspase-1 in Hypoxia group were higher than those in Normoxia group(P<0.05);Myocardial cell proliferation capacity in Hypoxia+Exo-NC group was higher than that in Hypoxia group,the apoptosis rate and the expression levels of ROS,NLRP3 and cleaved caspase-1 were lower than those in Hypoxia group(P<0.05);The proliferation ability of myocardial cells in Hypoxia+Exo-miR-486 group was higher than that in Hypoxia+Exo-NC group,and the apoptosis rate and the expression levels of ROS,NLRP3 and cleaved caspase-1 were lower than those in Hypoxia+Exo-NC group(P<0.05).Conclusion Exo from BMSC overexpressing with miR-486-5p can reduce hypoxic-induced ROS production and apoptosis of cardiomyocytes,and ameliorate hypoxic-induced slow proliferation of cardiomyocytes.

关 键 词：骨髓间充质干细胞 外泌体 心肌缺氧 miR-486-5p 凋亡 

分 类 号：R541.4[医药卫生—心血管疾病]