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目的研究血管内皮生长因子(vascularendothelial growth factor,VEGF)基因转染骨髓间充质干细胞(mesenchymalstem cells,MSCs)移植对缺血心肌的血管生成作用。方法于2004年5月至2005年8月取第四军医大学西京医院分离、培养Wistar大鼠的MSCs,用真核表达载体pcDNA3.1(-)/hVEGF165转染MSCs。45只近交系Wistar大鼠随机均分为转染组(MSCs/VEGF组)、对照组(MSCs组)、无血清培养基组(DMEM组),结扎前降支建立急性心肌梗死模型后在梗死区边缘区行5×106细胞移植,DMEM组行等量培养基注射。细胞移植前行CM-DiI标记。移植1个月后行心脏B超测量射血分数值,组织化学染色评价新生血管密度。结果培养的MSCs呈典型贴壁生长成纤维样外观,pcDNA3.1(-)/hVEGF165能有效转染大鼠MSCs,移植1个月后MSCs/VEGF组较其余各组左室射血分数(LVEF),再生血管密度明显增加,差异均有显著性(P<0.01)。结论VEGF基因转染MSCs移植能显著促进缺血心肌血管再生,进而改善心脏功能。
Objective To investigate the angiogenic effect of vascular endothelial growth factor (VEGF) gene transfected mesenchymal stem cells (MSCs) on ischemic myocardium. METHODS: MSCs of Wistar rats were isolated from Xijing Hospital of Fourth Military Medical University from May 2004 to August 2005, MSCs were transfected with eukaryotic expression vector pcDNA3.1 (-) / hVEGF165. Forty-five inbred Wistar rats were randomly divided into four groups: MSCs / VEGF group, MSCs group, DMEM group, anterior descending artery group, acute myocardial infarction model The marginal zone of infarction 5 × 106 cells were transplanted into the DMEM group, the same amount of medium injection. CM-DiI labeling before cell transplantation. One month after transplantation, cardiac B ultrasound was used to measure ejection fraction, and histochemical staining was used to evaluate neovascular density. Results MSCs cultured in vitro showed a typical appearance of adherent growth and fibroblast-like appearance. PcDNA3.1 (-) / hVEGF165 could effectively transfect rat MSCs. Compared with the other groups, MSCs / VEGF group showed a significant increase in left ventricular ejection fraction ), And the density of regenerating blood vessels increased significantly (P <0.01). Conclusion Transplantation of VEGF gene into MSCs can significantly promote the regeneration of ischemic myocardium and improve cardiac function.