单核巨噬细胞膜修饰iPSC-CM来源外泌体靶向修复损伤心脏的实验研究

Experimental Study on Targeted Repair of Injured Heart by iPSC-CM-derived Exosomes Modified by Monocyte-macrophage Membrane

目的利用单核巨噬细胞膜修饰人多潜能干细胞诱导心肌细胞(iPSC-CMs)来源的外泌体并探讨其心肌修复作用。方法使用高速离心法提取iPSC-CMs来源的外泌体,试剂盒提取单核巨噬细胞膜。采用脂质体挤出器融合外泌体和细胞膜,Western blot、透射电镜和纳米颗粒分析仪评价膜融合效果。采用CCK-8试验检测细胞活力,建立小鼠心肌梗死模型,隔天静脉注射M-iCM-EXs(1011个EXs/kg)3次;6 h后采用小动物活体光学成像系统评价M-iCM-EXs对损伤心脏的靶向性;28 d后采用Masson三色染色评估M-iCM-EXs治疗对心肌梗死后心室重构的作用和心动超声图评价M-iCM-EXs治疗后对的心功能变化。结果CCK-8试验证实M-iCM-EXs可提高OGD后细胞活力;小动物活体光学成像系统证实M-iCM-EXs对损伤心脏具有靶向性;Masson染色和心动超声图证实M-iCM-EXs治疗后能抑制小鼠心肌梗死后的心室重塑。结论单核巨噬细胞膜修饰iPSC-CM来源的外泌体具有靶向损伤心肌的能力,能更有效修复损伤心脏。

Objective To study and explore its myocardial repair effect.Methods The exosomes derived from iPSC-CMs were extracted by high-speed centrifugation,and the mononuclear macrophage membrane was extracted by the kit.The exosomes and cell membranes were fused by liposome extruder,and the membrane fusion effect was evaluated by Western blot,transmission electron microscopy and nanoparticle analyzer.CCK-8 assay was used to detect cell viability,and a mouse model of myocardial infarction was established.M-iCM-EXs(1011 EXs/kg)was intravenously injected every other day for 3 times.After 6 h,the targeting of M-iCM-EXs to injured heart was evaluated by small animal in vivo optical imaging system.After 28 days,Masson trichrome staining was used to evaluate the effect of M-iCM-EXs treatment on ventricular remodeling after myocardial infarction and echocardiography was used to evaluate the changes of cardiac function after M-iCM-EXs treatment.Results CCK-8 assay confirmed that M-iCM-EXs could improve cell viability after OGD.Small animal imaging technology confirmed the targeting of M-iCM-EXs to ischemic myocardial tissue.Masson trichrome staining test and echocardiography confirmed that M-iCM-EXs treatment could inhibit ventricular remodeling and improve cardiac function after MI in mice.Conclusion The iPSC-CMs-derived exosomes modified by monocyte-macrophage membrane have the ability to target damaged myocardium and can repair damaged heart more effectively.