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背景:骨髓间质干细胞和血管内皮细胞生长因子移植具有促进血管再生、改善心功能的作用,但是二者联合应用是否优于单独应用尚不清楚。目的:观察同种异体骨髓间质干细胞移植联合血管内皮生长因子基因转染对大鼠急性心肌梗死血管再生和心功能的影响。设计:大鼠骨髓间质干细胞培养采用单一样本观察;细胞移植和基因转染采用随机对照动物实验。单位:华中科技大学协和医院心内科,同济医学院心血管病研究所。材料:健康雄性Wistar大鼠94只。表达载体PAdTrack/VEGF165。方法:实验于2004-06/2005-06在同济医学院心血管病研究所实验室完成。①体外分离、纯化、培养大鼠骨髓间质干细胞,以5-溴-2’-脱氧尿苷标记细胞。②制备、抽提、纯化、鉴定质粒PAdTrack/VEGF165。③结扎冠状动脉建立急性心肌梗死模型2周后,随机将其分为4组,每组均为12只,干细胞+质粒组、干细胞组、质粒组、对照组,分别进行心肌内大鼠骨髓间质干细胞移植和/或VEGF165转染、DMEM注射。④4周后行免疫组织化学和超声心动图检查。主要观察指标:①各组大鼠梗死及缺血区免疫组织化学和苏木精-伊红染色检查。②血管计数。③超声心动图检查。结果:48只大鼠进入结果分析。①干细胞+质粒组和干细胞组梗死及缺血心肌处可见大量5-溴-2’-脱氧尿苷标记的移植细胞,其中缺血心肌处部分移植细胞分化为血管内皮细胞并形成新生毛细血管。②Ⅷ因子染色阳性的新生血管密度分布为干细胞+质粒组>质粒组>干细胞组>对照组(P均<0.01)。③细胞移植和基因转染治疗后室壁厚度和室壁运动幅度改善,射血分数值增加幅度为干细胞+质粒组>干细胞组>质粒组>对照组(P均<0.01)。结论:同种异体骨髓间质干细胞移植联合血管内皮生长因子基因转染能进一步增强大鼠梗死缺血区血管再生、改善室壁厚度和心功能。
BACKGROUND: Bone marrow mesenchymal stem cells and vascular endothelial growth factor transplantation have the effect of promoting blood vessel regeneration and improving cardiac function. However, it is unclear whether the combination of them is better than single application. OBJECTIVE: To observe the effects of allogenic bone marrow mesenchymal stem cell transplantation and vascular endothelial growth factor gene transfection on the vascular remodeling and cardiac function in rats with acute myocardial infarction. DESIGN: Rat bone marrow mesenchymal stem cells were cultured in a single sample; cell transplantation and gene transfection were randomized controlled animal experiments. Unit: Huazhong University of Science and Technology Union Hospital Cardiology, Tongji Medical College Institute of Cardiovascular Diseases. Materials: 94 healthy male Wistar rats. Expression vector PAdTrack / VEGF165. Methods: The experiment was performed in the laboratory of Institute of Cardiovascular Diseases, Tongji Medical College from June 2004 to June 2005. ① in vitro isolation, purification, culture of rat bone marrow mesenchymal stem cells, 5-bromo-2’-deoxyuridine labeled cells. ② Preparation, extraction, purification, identification of plasmid PAdTrack / VEGF165. ③ After 2 weeks of ligation of coronary artery, acute myocardial infarction model was randomly divided into 4 groups, each group was 12, stem cell + plasmid group, stem cell group, plasmid group and control group, Stem cell transplantation and / or VEGF165 transfection, DMEM injection. ④ 4 weeks after the line immunohistochemistry and echocardiography. MAIN OUTCOME MEASURES: ①Immunohistochemistry and hematoxylin-eosin staining of infarction and ischemic area in each group were performed. ② blood vessel count. ③ echocardiography. Results: 48 rats entered the result analysis. ① A large number of 5-bromo-2’-deoxyuridine-labeled transplanted cells were observed in infarcted and ischemic myocardium of stem cell + plasmid group and stem cell group, in which some of the transplanted cells in ischemic myocardium differentiated into vascular endothelial cells and formed new capillaries. The Ⅷ factor positive neovascularization density distribution was stem cell + plasmid group> plasmid group> stem cell group> control group (all P <0.01). ③ The cell wall thickness and wall motion amplitude were improved after transplanting and gene transfection. The increasing rate of ejection fraction was stem cell + plasmid group> stem cell group> plasmid group> control group (all P <0.01). CONCLUSION: Transplantation of allogenic bone marrow mesenchymal stem cells combined with vascular endothelial growth factor gene transfection can further enhance the angiogenesis of ischemic infarction in rats and improve the wall thickness and cardiac function.