金納多注射液對大鼠急性心肌梗死后hAD-MSCs移植的影響
本文選題:金納多注射液 + 急性心肌梗死; 參考:《北京中醫(yī)藥大學(xué)》2017年碩士論文
【摘要】:目的:研究金納多注射液對人羊膜間充質(zhì)干細(xì)胞移植治療大鼠急性心肌梗死的影響。方法:本研究選擇40只SD(Sprague-Dawley)雄性大鼠隨機(jī)分為4組:金納多注射液+干細(xì)胞組(n=10),干細(xì)胞組(n=10),模型組(n=10),假手術(shù)組(n=10)。結(jié)扎大鼠左冠狀動脈前降支建立急性心肌梗死(acute myocardial infarction,AMI)模型。金納多注射液+干細(xì)胞組、干細(xì)胞組在造模后當(dāng)天,將分離純化的第三代hAD-MSCs,經(jīng)心外膜下注入大鼠體內(nèi)心肌梗死區(qū)及周邊組織。模型組和假手術(shù)組分別注入等量的無血清DMEM/F12培養(yǎng)基。金納多注射液+干細(xì)胞組在hAD-MSCs移植前1周及后1周連續(xù)予金納多注射按7.35mg/(Kg·d)進(jìn)行灌胃。干細(xì)胞組、模型組、假手術(shù)組用生理鹽水灌胃。于hAD-MSCs移植后1周各組進(jìn)行超聲心動圖檢查。移植后1周將大鼠處死進(jìn)行組織病理學(xué)、免疫組織化學(xué)等檢測,以觀察金納多注射液對心肌梗死區(qū)心室重構(gòu)和心功能的影響。結(jié)果:①所分離的hAD-MSCs具有典型的間充質(zhì)干細(xì)胞表型特征,高表達(dá)間充質(zhì)干細(xì)胞特異性蛋白STRO-1和Vimentin,不表達(dá)上皮細(xì)胞標(biāo)記的CK-19,說明分離得到的HAD-MSCs具有間充質(zhì)干細(xì)胞的特性;②與模型組相比,金納多注射液+干細(xì)胞組、干細(xì)胞組大鼠左室舒張末內(nèi)徑顯著變小,金納多注射液+干細(xì)胞組與干細(xì)胞組比較無顯著差異。與模型組比較,金納多注射液+干細(xì)胞組、干細(xì)胞組大鼠左室射血分?jǐn)?shù)和左室短軸縮短率顯著高于模型組(P0.01);與干細(xì)胞組比較,金納多注射液+干細(xì)胞組大鼠左室射血分?jǐn)?shù)和左室短軸縮短率改善更顯著(P0.01)。③與模型組相比,金納多注射液+干細(xì)胞組、干細(xì)胞組心肌細(xì)胞變性較輕,新生血管形成較多;金納多注射液+干細(xì)胞組與干細(xì)胞組相比,炎性細(xì)胞浸潤較少,心肌細(xì)胞變性較輕,新生血管形成較多;④金納多注射液+干細(xì)胞組、干細(xì)胞組心肌梗死區(qū)及周邊組織的纖維化程度較模型組降低;金納多注射液+干細(xì)胞組心肌梗死區(qū)及周邊組織的纖維化程度較干細(xì)胞組降低;⑤金納多注射液+干細(xì)胞組、干細(xì)胞組心肌梗死區(qū)及周邊組織的細(xì)胞凋亡較模型組顯著減少;金納多注射液+干細(xì)胞組較干細(xì)胞組心肌梗死區(qū)及周邊組織的的細(xì)胞凋亡顯著減少。結(jié)論:金納多注射液聯(lián)合hAD-MSCs可改善心肌梗死區(qū)及周邊區(qū)域的微環(huán)境,減輕心肌組織纖維化,減少心肌細(xì)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.
【學(xué)位授予單位】:北京中醫(yī)藥大學(xué)
【學(xué)位級別】:碩士
【學(xué)位授予年份】:2017
【分類號】:R542.22
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