血管内皮生长因子转染内皮祖细胞治疗大鼠缺血后肢的研究

目的 使用血管内皮生长因子(VEGF)转染内皮祖细胞(EPC)治疗大鼠缺血后肢,观察EPC、VEGF转染EPC对大鼠缺血后肢的新生血管和肢体成活的影响.方法 制作SD大鼠后肢缺血模型,将动物随机分为3组,每组6只.将构建的VEGF基因真核表达载体转染入骨髓来源的EPCs后通过尾静脉注射人大鼠体内,并与使用磷酸盐缓冲液(PBS)或EPC的动物进行比较,观察转染VEGF的EPCs在缺血部位的聚集和形成新生血管的情况.结果 (1)动物总残肢率比较,CELL组、VEGF组较PBS组明显增加的肢体恢复率(P<0.05),CELL组肢体恢复率较VEGF组差(P<0.05).(2)毛细血管密度与PBS组比较,各时间点中CELL、VEGF组MVD均明显增多(P<0.05).(3)缺血肢体VEGFa的表达:VEGF组的VEGF蛋白表达较PBS组、CELL组、明显增多(P<0.05);(4)手术后7、14、28 d,与PBS对照组比较,CELL、VEGF组细胞的血流灌注有较大程度的恢复(P<0.01).结论 VEGFa基因转染EPCs对缺血部位的血管新生有重要影响,联合应用VEGFa基因和EPCs治疗缺血后肢有较好的协同作用.

Study on Treatment of EPC Transfected with VEGFa on Rat Ischemic Hind-limb

Objective Using EPC transfected VEGFa to treat rat ischemic hind-limb,thus to explore the effect of EPC,EPC transfected with VEGF on new vessels of rat ischemic hind-limb and limb survival. Methods 54 models of SD rat ischemic hind-limb were established, and randomly divided into three groups. Each group has six rats. Eukaryotic expression vector of VEGF was transfected into EPCs from marrow,which then was injected into rats via vena caudalis. Compared them with animals applied PBS or EPC,and observed how EPCs transfected with VEGFa gene gathered at the ischemic part and facilitated angiogenesis. Results (1) As for the comparison of general limb disability, limb restoration rate of Group EPC,VEGF were obviously higher than that of Group PBS (P<0. 05). (2) Micro Vessel Density (MVD):compared with Group PBS,there was a marked increase in MVD of Group EPC, VEGF at each time point (P<0. 05). 14 and 21 days after operation, MVD of Group VEGF was higher than that of Group EPC (P<0.05). (3) Expression of VEGFa in ischemic limbs:compared with Group PBS,EPC, VEGF protein expression of Group VEGF was significantly increased (P<0. 05). Compared with Group PBS,and,there was obvious increase in VEGF protein expression in Group EPC (P<0.05). (4) Seven days after operation,rat limb perfusion in all groups was reduced:14,21 days after operation,compared with PBS control group,there was notable blood perfusion recovery in Group EPC and VEGF (P<0.01). Blood perfusion of Group EPC was less than that of Group VEGF (P<0.05 ). Conclusion EPCs transfected with VEGFa gene had a significant impact on angiogenesis at ischemic part. Combined application of VEGFa gene to the treatment of ischemic hind-limb has a good synergistic effect.