过表达血小板衍生生长因子受体β的小鼠内皮祖细胞有助于损伤后血管的再内皮化

Platelet derived growth factor receptor β over-expression in endothelial progenitor cells promote reendothelialization after vascular injury

目的 观察高表达血小板衍生生长因子受体β（PDGFR-β）的内皮祖细胞（EPC）修复损伤血管的功能.方法 采用密度梯度离心法获取小鼠脾脏单个核细胞,使用含血管内皮细胞的专用培养基诱导脾源性EPC,使用基因转染的方法获取高表达PDGFR-β的EPC,并将其回输至脾切除后的颈动脉损伤模型中,以伊文思蓝染色评估血管再内皮化程度.结果 荧光双染鉴定显示培养的EPC阳性率在90％以上,荧光显微镜观察表明质粒转染效率为50％～ 60％,转染后EPC高表达PDGFR-β,伊文思蓝染色结果显示EPC尾静脉移植,有助于损伤后血管的再内皮化,PDGFR-β过表达EPC移植效果优于单纯EPC移植（P＜0.01）.结论 体外培养脾源性EPC切实可行,使用基因转染的方法可以有效上调PDGFR-β表达,而过表达PDGFR-β的EPC显著促进小鼠颈动脉损伤早期血管再内皮化.

Objective To observe the effect of platelet derived growth factor receptor （PDGFR-β） transfected endothelial progenitor cells （EPCs） on vascular regeneration. Methods Spleen-derived mononuclear cells （MNCs） were isolated using density gradient centrifugation and induced with special culture medium. EPCs transfection was performed with LipofectamineTM 2000 reagent according to the instruction manual. Carotid artery injury was induced in splenectomized mice. EPCs were injected by tail vein immediately and at 24 h after endothelial injury of the carotid artery. Evans blue staining was performed to evaluate reendothelialization at 7 days after endothelial injury of the carotid artery. Results Most adherent cells were LDL and UEA-I double positive. Laser scanning confocal microscopy showed that transfection efficiency was about 50% -60%. The reendothelialized area in the PDGFR-IS-EPCs group was significantly larger than that in EGFP-EPCs group. Conclusion Transplantation of PDGFR-β over-expressed EPCs can promote reendothelialization in the early phase after carotid artery injury.