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目的:将编码有血管内皮生长因子基因的腺病毒(Ad.VEGF)转染诱导培养的骨髓内皮祖细胞(EPCs),移植于大鼠缺血心肌,评价其对心功能的保护和促血管再生作用。方法:Ficoll离心分离大鼠骨髓单个核细胞,诱导培养EPCs;在50倍的转染倍数(病毒数/靶细胞)下,用Ad.VEGF转染诱导的EPCs。然后将其移植于结扎冠状动脉前降支的Lewis大鼠缺血心肌内(组Ⅰ),同时设置单纯细胞移植组(组Ⅱ)和基因治疗组(组Ⅲ),以注射磷酸盐缓冲液的大鼠为对照组(组Ⅳ)。术后4周通过血流动力学指标评价其对心功能的保护作用;另外通过免疫组化染色观察移植细胞存活、分化和血管再生情况。结果:骨髓EPCs移植于缺血心肌4周后,缺血区的移植细胞部分分化为血管内皮细胞,组Ⅰ心功能明显好于组Ⅱ、组Ⅲ和组Ⅳ(P<0.01),在促血管新生方面,组Ⅰ也明显优于以上3组[组Ⅰ(32.2±3.5)、组Ⅱ(17.6±2.7)、组Ⅲ(19.4±3.3)、组Ⅳ(5.9±1.2)个/高倍视野]。结论:转染Ad.VEGF后的EPCs移植于缺血心肌后,可促进缺血心肌再血管化,更好地保护和改善心脏功能。
OBJECTIVE: To transfect bone marrow endothelial progenitor cells (EPCs) induced by Ad.VEGF encoding vascular endothelial growth factor (VEGF) into ischemic myocardium of rats to evaluate the protection of cardiac function and angiogenesis effect. Methods: EPCs were isolated from rat bone marrow mononuclear cells by Ficoll centrifugation, and EPCs were transfected with Ad.VEGF at a multiplication factor of 50 (virus / target cells). The cells were then transplanted into the ischemic myocardium of Lewis rats ligation of the anterior descending coronary artery (Group Ⅰ), and the cell transplantation group (Group Ⅱ) and the gene therapy group (Group Ⅲ) were set at the same time to inject phosphate buffered saline Rats as control group (Group IV). Four weeks after surgery, the hemodynamic parameters were used to evaluate the protective effect on cardiac function. The survival, differentiation and angiogenesis were observed by immunohistochemistry. Results: After transplantation of bone marrow EPCs into the ischemic myocardium for 4 weeks, the transplanted cells in the ischemic area partially differentiated into vascular endothelial cells. The cardiac function of group Ⅰ was significantly better than that of group Ⅱ, group Ⅲ and group Ⅳ (P <0.01) In newborn, group Ⅰ was also significantly better than the above three groups [group Ⅰ (32.2 ± 3.5), group Ⅱ (17.6 ± 2.7), group Ⅲ (19.4 ± 3.3), group Ⅳ (5.9 ± 1.2) / high power field]. CONCLUSION: EPCs transfected with Ad.VEGF can promote the revascularization of ischemic myocardium after transplanted into ischemic myocardium, so as to better protect and improve cardiac function.