血管内皮生长因子基因修饰骨髓间充质干细胞移植心肌梗死大鼠:高压氧干预促进治疗性血管生成

摘要

背景:骨髓间充质干细胞移植入缺血心肌后梗死区周边心肌的缺血、缺氧、炎性反应、细胞凋亡等病理变化严重影响到移植骨髓间充质干细胞的存活,而高压氧能显著改善其缺氧状态.目的:对心肌梗死模型大鼠缺血心肌局部移植血管内皮生长因子修饰的骨髓间充质干细胞,在移植后对大鼠进行高压氧干预,探讨其对骨髓间充质干细胞移植后所获得的治疗性血管生成效应的影响.方法:将真核表达载体pcDNA3.1(-)/人血管内皮生长因子165转染大鼠骨髓间充质干细胞,再移植至心肌梗死模型大鼠的缺血区组织,移植前细胞行CM-DiI标记.移植后2周每日对大鼠行高压氧治疗干预.移植1个月后行心脏B超测量其左室射血分数,组织化学苏木精-伊红染色和Ⅷ因子染色并评价新生血管密度.结果与结论:CM-DiI能有效标记骨髓间充质干细胞,标记率近100%.细胞移植1个月后高压氧干预的大鼠射血分数、再生血管密度较均明显增高(P < 0.05).证实,高压氧干预能显著促进移植血管内皮生长因子基因修饰的骨髓间充质干细胞的心肌梗死大鼠所获得的治疗性血管生成作用,对心脏功能有明显改善作用.%BACKGROUND: The pathological changes of bone marrow mesenchymal stem cells after transplantation into peri-infarct myocardium including ischemia, hypoxia, inflammation, and cell apoptosis greatly influence the survival of transplanted bone marrow mesenchymal stem cells, while hyperbaric oxygenation can significantly improve the hypoxic state.rnOBJECTIVE: To investigate the effects of hyperbaric oxygenation for therapeutic angiogenesis acquired by transplantation of vascular endothelial growth factor gene modified bone marrow mesenchymal stemcells in rats with myocardial infarction.rnMETHODS: Rat bone marrow mesenchymal stem cells transfected with eukaryotic expression vector pcDNA3.1(-)/VEGF165 were transplanted into the ischemic tissue of a rat model of myocardial infarction. Priorto transplantation, rat bone marrow mesenchymal stem cells were labeled with CM-Dil. Within 2 weeks after transplantation, hyperbaric oxygenation was performed daily. At 1 month after transplantation, the left ventricular ejection fraction was measured by ultrasonic cardiogram, the density of newly formed vessels was determined by histochemical, hematoxylin-eosin, and V1H factor staining.rnRESULTS AND CONCLUSION: CM-Dil could label nearly 100% bone marrow mesenchymal stem cells. At 1 month after transplantation, the left ventricular ejection fraction and the density of newly formed vessels in rats subjected to hyperbaric oxygenation were significantly increased (P < 0.05). Hyperbaric oxygenation after cell transplantation results in remarkable therapeutic angiogenesis and improves cardiac function by transplantation of vascular endothelial growth factor gene-modified bone marrow mesenchymal stem cells.