金纳多注射液对大鼠急性心肌梗死后hAD-MSCs移植的影响

发布时间：2018-04-28 01:52

本文选题：金纳多注射液 + 急性心肌梗死　；参考：《北京中医药大学》2017年硕士论文

【摘要】：目的:研究金纳多注射液对人羊膜间充质干细胞移植治疗大鼠急性心肌梗死的影响。方法:本研究选择40只SD(Sprague-Dawley)雄性大鼠随机分为4组:金纳多注射液+干细胞组(n=10),干细胞组(n=10),模型组(n=10),假手术组(n=10)。结扎大鼠左冠状动脉前降支建立急性心肌梗死(acute myocardial infarction,AMI)模型。金纳多注射液+干细胞组、干细胞组在造模后当天,将分离纯化的第三代hAD-MSCs,经心外膜下注入大鼠体内心肌梗死区及周边组织。模型组和假手术组分别注入等量的无血清DMEM/F12培养基。金纳多注射液+干细胞组在hAD-MSCs移植前1周及后1周连续予金纳多注射按7.35mg/(Kg·d)进行灌胃。干细胞组、模型组、假手术组用生理盐水灌胃。于hAD-MSCs移植后1周各组进行超声心动图检查。移植后1周将大鼠处死进行组织病理学、免疫组织化学等检测,以观察金纳多注射液对心肌梗死区心室重构和心功能的影响。结果:①所分离的hAD-MSCs具有典型的间充质干细胞表型特征,高表达间充质干细胞特异性蛋白STRO-1和Vimentin,不表达上皮细胞标记的CK-19,说明分离得到的HAD-MSCs具有间充质干细胞的特性;②与模型组相比,金纳多注射液+干细胞组、干细胞组大鼠左室舒张末内径显著变小,金纳多注射液+干细胞组与干细胞组比较无显著差异。与模型组比较,金纳多注射液+干细胞组、干细胞组大鼠左室射血分数和左室短轴缩短率显著高于模型组(P0.01);与干细胞组比较,金纳多注射液+干细胞组大鼠左室射血分数和左室短轴缩短率改善更显著(P0.01)。③与模型组相比,金纳多注射液+干细胞组、干细胞组心肌细胞变性较轻,新生血管形成较多;金纳多注射液+干细胞组与干细胞组相比,炎性细胞浸润较少,心肌细胞变性较轻,新生血管形成较多;④金纳多注射液+干细胞组、干细胞组心肌梗死区及周边组织的纤维化程度较模型组降低;金纳多注射液+干细胞组心肌梗死区及周边组织的纤维化程度较干细胞组降低;⑤金纳多注射液+干细胞组、干细胞组心肌梗死区及周边组织的细胞凋亡较模型组显著减少;金纳多注射液+干细胞组较干细胞组心肌梗死区及周边组织的的细胞凋亡显著减少。结论:金纳多注射液联合hAD-MSCs可改善心肌梗死区及周边区域的微环境,减轻心肌组织纤维化,减少心肌细胞凋亡,改善心功能。
[Abstract]:Aim: to study the effect of Ginaton injection on the transplantation of human amniotic mesenchymal stem cells in acute myocardial infarction in rats. Methods: forty SD Sprague-Dawley (SD) male rats were randomly divided into 4 groups: Jinnado injection stem cell group, model group and sham operation group. Acute myocardial infarction (AMI) model was established by ligating the left anterior descending coronary artery in rats. The third generation of hAD-MSCswere isolated and purified and injected into the myocardial infarction area and surrounding tissues of rats under epicardial membrane on the same day after the establishment of the model. The model group and the sham operation group were injected with the same amount of serum-free DMEM/F12 medium. Ginado injection stem cells group was treated with 7.35mg/(Kg d 1 week before and 1 week after hAD-MSCs transplantation. Stem cells group, model group and sham operation group were treated with normal saline. Echocardiography was performed 1 week after hAD-MSCs transplantation. One week after transplantation, the rats were sacrificed for histopathological and immunohistochemical examination to observe the effects of Ginado injection on ventricular remodeling and cardiac function in myocardial infarction area. Results the hAD-MSCs isolated by 1: 1 had typical phenotypic characteristics of mesenchymal stem cells, high expression of mesenchymal stem cell specific proteins STRO-1 and Vimentin, and no expression of epithelial cell labeled CK-19, indicating that the isolated HAD-MSCs possessed the characteristics of mesenchymal stem cells. 2Compared with the model group, the left ventricular end-diastolic diameter in the stem cell group was significantly smaller than that in the model group, but there was no significant difference between the stem cell group and the stem cell group. Compared with the model group, the left ventricular ejection fraction and the shortening rate of the left ventricular short axis in the stem cell group were significantly higher than those in the model group, and those in the stem cell group were significantly higher than those in the model group, and those in the stem cell group were significantly higher than those in the model group. The left ventricular ejection fraction (LVEF) and the shortening rate of the short axis of left ventricle were improved more significantly in the stem cell group than in the model group. Compared with the model group, the myocardial cell degeneration and angiogenesis in the stem cell group were lighter and the neovascularization rate was higher in the stem cell group than in the model group. Compared with the stem cell group, the inflammatory cell infiltration was less, the degeneration of myocardial cells was lighter, and the neovascularization was more frequent in the stem cell group than in the stem cell group. The degree of fibrosis in myocardial infarction area and surrounding tissue in stem cell group was lower than that in model group, and the fibrosis degree in myocardial infarction area and surrounding tissue in Ginaton injection stem cell group was lower than that in stem cell group, and that in Jinnadio injection stem cell group was lower than that in stem cell group. The apoptosis of myocardial infarction area and peripheral tissue in stem cell group was significantly lower than that in model group, and that in stem cell group was significantly lower than that in stem cell group. Conclusion: Ginado injection combined with hAD-MSCs can improve the microenvironment of myocardial infarction area and surrounding area, alleviate myocardial fibrosis, reduce myocardial cell apoptosis and improve cardiac function.
【学位授予单位】：北京中医药大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R542.22

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