Zou Liyuan, Ma Xiaokun, Lin Shuo, Wu Bingyuan, Chen Yang, Peng Chaoquan
Department of Prevention and Health Care, The Third Affiliated Hospital of Sun Yat-sen University, Guangzhou, Guangdong 510630, P.R. China.
Department of Medical Oncology, The Third Affiliated Hospital of Sun Yat-sen University, Guangzhou, Guangdong 510630, P.R. China.
Exp Ther Med. 2019 Oct;18(4):2574-2582. doi: 10.3892/etm.2019.7874. Epub 2019 Aug 9.
Exosomes have been demonstrated to be effective in the treatment of a variety of cardiac disorders. However, the effects of mesenchymal stem cell (MSC) exosomes on myocardial infarction is yet to be determined. The current study aimed to investigate the potential therapeutic effects of MSC exosomes on myocardial injuries that are caused by myocardial infarction. MSCs were isolated from rat bone marrow and were used for exosome enrichment using culture medium. Confirmation that MSCs and exosomes had been successfully extracted was performed using flow cytometry, electron microscopy and western blot analysis. A rat myocardial ischemia reperfusion (I/R) model was established by ligation of the left anterior descending coronary artery. Rat myocardial injuries were determined using 2,3,5-triphenyltetrazolium chloride, Masson and TUNEL staining. H9c2 cell proliferation, apoptosis and migration were analyzed using 5-ethynyl-2'-deoxyuridine, Hoechst staining, flow cytometry and Transwell assays. Marker gene expression was evaluated using reverse transcription-quantitative PCR, western blot analysis and immunofluorescence. Rat MSC exosomes were revealed to suppress myocardial injury and the myocardiocyte functions that were induced by I/R. The results also demonstrated decreased apoptotic protease activating factor-1 and increased autophagy-related protein 13 expression. The H9c2 cell proliferation and migration inhibition, as well as cell apoptosis during hypoxia-reoxygenation (H/R), were suppressed by rat MSC exosomes, with an alteration of the expression of apoptotic and autophagic genes also being demonstrated. The application of autophagy inhibitor 3-methyladenine significantly mitigated the effect of exosomes on H9c2 cell proliferation and apoptosis, which were induced by H/R. Rat MSC exosomes inhibited myocardial infarction pathogenesis, possibly by regulating autophagy.
外泌体已被证明在治疗多种心脏疾病方面有效。然而，间充质干细胞（MSC）外泌体对心肌梗死的影响尚未确定。当前研究旨在探讨MSC外泌体对心肌梗死所致心肌损伤的潜在治疗作用。从大鼠骨髓中分离出MSC，并使用培养基进行外泌体富集。通过流式细胞术、电子显微镜和蛋白质印迹分析来确认MSC和外泌体已成功提取。通过结扎左冠状动脉前降支建立大鼠心肌缺血再灌注（I/R）模型。使用氯化三苯基四氮唑、Masson染色和TUNEL染色来确定大鼠心肌损伤。使用5-乙炔基-2'-脱氧尿苷、Hoechst染色、流式细胞术和Transwell实验分析H9c2细胞增殖、凋亡和迁移。使用逆转录定量PCR、蛋白质印迹分析和免疫荧光评估标志物基因表达。结果显示，大鼠MSC外泌体可抑制I/R诱导的心肌损伤和心肌细胞功能。结果还表明凋亡蛋白酶激活因子-1减少，自噬相关蛋白13表达增加。大鼠MSC外泌体抑制了缺氧复氧（H/R）期间H9c2细胞的增殖和迁移以及细胞凋亡，同时也显示出凋亡和自噬基因表达的改变。自噬抑制剂3-甲基腺嘌呤的应用显著减轻了外泌体对H/R诱导的H9c2细胞增殖和凋亡的影响。大鼠MSC外泌体可能通过调节自噬来抑制心肌梗死的发病机制。
