miRNA-210修饰的MSCs治疗心肌缺血再灌注损伤模型大鼠的实验研究

Experimental Study of miRNA-210 modified MSCs in the treatment of myocardial Ischemiaischemia-reperfusion injury in rats

目的:探讨微小RNA210(miRNA-210)修饰的骨髓间充质干细胞(MSCs)对心肌缺血再灌注损伤(MIRI)模型大鼠的治疗作用。方法:选取1只SD大鼠将其处死,分离其下肢胫骨和股骨,采用全骨髓贴壁法培养MSCs,构建miRNA-210修饰的MSCs。选取40只SD大鼠分为假手术组、模型组、MSCs组、miRNA-210+MSCs组,每组10只,采用结扎左冠状动脉前降支法制备心肌缺血再灌注模型,造模成功后MSCs组尾静脉注射50μL MSCs悬液,miRNA-210+MSCs组尾静脉注射50μL miRNA-210修饰的MSCs悬液,假手术组和模型组注射等量生理盐水。造模第10天,比较各组心肌梗死面积、心肌组织形态学变化、心肌细胞凋亡率、miRNA-210表达量。结果:模型组心肌梗死面积及心肌细胞凋亡率高于假手术组(P<0.05);MSCs组心肌梗死面积及心肌细胞凋亡率低于模型组(P<0.05);miRNA-210+MSCs组心肌梗死面积及心肌细胞凋亡率低于MSCs组(P<0.05);miRNA-210+MSCs组心肌梗死面积及心肌细胞凋亡率高于假手术组(P<0.05);模型组心肌组织中miRNA-210表达量高于假手术组(P<0.05);MSCs组心肌组织中miRNA-210表达量较模型组无明显差异(P>0.05);miRNA-210+MSCs组心肌组织中miRNA-210表达量高于MSCs组、模型组、假手术组(P<0.05);HE染色显示miRNA-210+MSCs组大鼠心肌组织形态基本正常,胞浆染色较均匀,心肌纤维排列基本整齐,炎性细胞浸润及间质水肿较模型组和MSCs组明显改善。结论:miRNA-210修饰的MSCs可以抑制心肌缺血再灌注损伤模型大鼠心肌细胞凋亡,减少大鼠心肌梗死面积,改善大鼠心肌组织病理损伤,从而对心肌缺血再灌注损伤起到一定治疗作用。

Objective:To investigate the effect of microRNA210(miRNA-210)modified bone marrow mesenchymal stem cells(MSCs)on myocardial ischemia-reperfusion injury(MIRI)model rats.Methods:The tibia and femur of the lower extremity of 1 SD rat were isolated.MSCs could be obtained by whole bone marrow adherent method,and were modified by miRNA-210.Another Forty 40 SD rats were divided into sham operation group,model group,MSCs group and miRNA-210+MSCs group,with 10 rats in each group.Myocardial ischemia-reperfusion model was established by ligating the anterior descending branch of left coronary artery.After model establishment,50μL of MSCs suspension was injected into the tail vein of MSCs group,50μL of miRNA-210 modified MSCs suspension was injected into the tail of miRNA-210+MSCs group,and the same amount of normal saline was injected into sham operation group and model group.On the 10th day,myocardial infarction area,myocardial histomorphology,apoptosis rate and miRNA-210 expression were compared.Results:The myocardial infarction area and myocardial apoptosis rate among four groups ranking in a descending order were model group,MSCs group,miRNA-210+MSCs group and sham operation group,with statistic significant difference(P<0.05).The expression of miRNA-210 in the myocardial tissue of the model group was significantly higher than that in the sham operation group(P<0.05).The expression of miRNA-210 in myocardial tissue of MSCs group was not significantly different from that of model group(P>0.05).The expression of miRNA-210 in myocardial tissue of miRNA-210+MSCs group was significantly higher than that of MSCs group,model group and sham operation group(P<0.05).He HE staining showed that the myocardial morphology of mirna-210+MSCs group was basically normal,the cytoplasmic staining was more uniform,and the arrangement of myocardial fibers was basically orderly,;moreover,the inflammatory cell infiltration and interstitial edema were significantly improved compared with those of model group and MSCs group.Conclusion:The miRNA-210 modified MSCs can inhibit the apoptosis of myocardial cells,reduce the myocardial infarction area and improve the pathological injury of myocardial tissue in rats with myocardial ischemia-reperfusion injury.