骨髓间充质干细胞联合血管内皮生长因子基因治疗肢体缺血的实验研究

Bone marrow mesenchymal stem cells combined with VEGF gene for the treatment of limb ischemia in rabbits

目的 观察骨髓间充质干细胞（bone mesenchymal stem cell,BMSC）联合血管内皮生长因子（vascular endothelial growth factor,VEGF）基因治疗对家兔肢体缺血模型的疗效.方法 切除新西兰兔右后肢全长股浅动脉并结扎股深动脉以建立兔后肢缺血模型,随机分为空质粒对照组（EP组）、骨髓间充质干细胞组（BMSC组）、VEGF基因治疗组（VEGF组）及联合治疗组（BV组）,每组各8只.分别于治疗后28 d及30 d进行动脉造影及VEGF免疫组化染色.结果 EP组、BMSC组及VEGF组的新生血管计数组间比较差异无统计学意义（P＞0.05）.BV组的新生血管计数较其余3组明显增加,差异有统计学意义（F=35.47,P＜O.01）.BMSC组及VEGF组的VEGF免疫组化染色呈阳性表达,与EP组比较差异有统计学意义（F=764.32,P＜0.01）.BV组的VEGF免疫组化染色呈强阳性表达,与其余3组比较差异有统计学意义（F =764.32,P＜0.01）.结论 BMSC联合VEGF基因治疗兔肢体缺血可使VEGF获得稳定而有效的表达,从而改善肢体缺血.

Objective To evaluate bone marrow mesenchymal stem cells combined with VEGF gene in the treatment of limb ischemia in rabbits.Methods The right hind limb ischemia model of New Zealand rabbit was established by superficial femoral artery excision and deep femoral artery ligation.Rabbits then were divided randomly into 4 groups： empty plasmid control group（EP group）,bone marrow mesenchymal stem cells group（BMSC group）,VEGF gene therapy group（VEGF group）,combination bone mesenchymal stem cells and VEGF gene therapy group（BV group）.There were 8 rabbits in each group.Angiogenesis was detected by arteriography on day 28 after treatment and expression of VEGF was detected by immunohistochemical staining on day 30 after treatment.Results There were no differences of collateral vessel count between the EP group,BMSC group and VEGF group.The collateral vessel count in BV group was higher than that of the other three groups.Immunohistochemistry of VEGF showed that the integrated optical density（IOD）in BMSC and VEGF groups increased significantly compared with the EP group; the IOD in BV group was the highest compared with the other three groups.Conclusions Combination bone marrow mesenchymal stem cells and VEGF gene in the treatment of limb ischemia in rabbits can obtain stable and effective expression of VEGF along with significant improvement of limb ischemia.