Bone marrow mesenchymal stem cell-secreted exosomes carrying microRNA-125b protect against myocardial ischemia reperfusion injury via targeting SIRT7
Bone marrow mesenchymal stem cell-secreted exosomes carrying microRNA-125b protect against myocardial ischemia reperfusion injury via targeting SIRT7
复制标题
骨髓间充质干细胞分泌的携带microRNA-125b的外泌体通过靶向SIRT7保护心肌缺血再灌注损伤
DOI：
10.1007/s11010-019-03671-z
复制
发表时间：
2020-02-01
影响因子：
4.3
通讯作者：
Fu, Guosheng
中科院分区：
文献类型：
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作者：
Chen, Qi;Liu, Yu;Fu, Guosheng
microRNA-125 b（miR-125 b）可减少心肌梗死面积，抑制心肌缺血再灌注损伤（I/R）。本研究旨在探讨携带miR-125 b的骨髓间充质干细胞（BMSC）来源的exosomes对I/R大鼠的影响。结扎大鼠冠状动脉左前降支（LAD）建立心肌I/R模型。将大鼠随机分为I/R组和Sham组。将Lv-cel-miR-67（对照）或Lv-miR-125 b转染到BMSC中。从转染的BMSC中提取外泌体，并分别命名为BMSC-Exo-67、BMSC-Exo-125 b和BMSC-Exo。MTT法检测心肌细胞存活率，流式细胞术检测心肌细胞凋亡。Western blot检测细胞凋亡蛋白的表达，ELISA检测炎症因子的水平。利用StarBase 3. 0软件预测miR-125 b与SIRT 7的靶向关系，并通过双荧光素酶报告基因分析进行验证。采用qRT-PCR、免疫组化和Western blot方法检测SIRT 7在I/R大鼠心肌组织中的表达。BMSC来源的外泌体被成功分离并通过TEM和CD 9和CD 63的阳性表达鉴定。miR-125 b在I/R心肌组织和细胞中表达下调。BMSC-Exo-125 b显著上调I/R心肌细胞中miR-125 b的表达。BMSC-Exo-125 b干预可显著提高I/R心肌细胞的存活率，降低凋亡率，下调Bax和caspase-3表达，上调Bcl-2表达，降低IL-1β、IL-6和TNF-α水平。SIRT 7是miR-125 b的靶点，BMSC-Exo-125 b显著下调心肌细胞中SIRT 7的表达。BMSC-Exo-125 b注射可减轻I/R大鼠心肌组织病理损伤，下调SIRT 7表达。携带miR-125 b的BMSC衍生的外泌体通过靶向SIRT 7保护心肌I/R。
MicroRNA-125b (miR-125b) reduces myocardial infarct area and restrains myocardial ischemia reperfusion injury (I/R). In this study, we aimed to investigate the effect of bone marrow mesenchymal stem cell (BMSC)-derived exosomes carrying miR-125b on I/R rats. The myocardial I/R model in rats was constructed by ligation of the left anterior descending coronary artery (LAD). Rats were randomly divided into I/R and Sham group. Lv-cel-miR-67 (control) or Lv-miR-125b was transfected into BMSCs. Exosomes were extracted from transfected BMSCs, and separately named BMSC-Exo-67, BMSC-Exo-125b, and BMSC-Exo. MTT assay and flow cytometry were used to detect the viability and apoptosis of I/R myocardium cells, respectively. The expression of cell apoptosis proteins and the levels of inflammatory factors were examined by Western blot and ELISA assay, respectively. The target relationship between miR-125b and SIRT7 was predicted by using StarBase3.0, and was confirmed by using dual-luciferase reporter gene assay. qRT-PCR, immunohistochemistry staining, and Western blot were used to evaluate the expression of SIRT7 in myocardium tissues in I/R rats. BMSC-derived exosomes were successfully isolated and identified by TEM and positive expression of CD9 and CD63. The expression of miR-125b was down-regulated in I/R myocardium tissues and cells. BMSC-Exo-125b significantly up-regulated miR-125b in I/R myocardium cells. The intervention of BMSC-Exo-125b significantly increased the cell viability, decreased the apoptotic ratio, down-regulated Bax and caspase-3, up-regulated Bcl-2, and decreased the levels of IL-1β, IL-6, and TNF-α in I/R myocardium cells. SIRT7 was a target of miR-125b, and BMSC-Exo-125b significantly down-regulated SIRT7 in myocardium cells. In addition, the injection of BMSC-Exo-125b alleviated the pathological damages and down-regulated SIRT7 in myocardium tissues of I/R rats. BMSC-derived exosomes carrying miR-125b protected against myocardial I/R by targeting SIRT7.