血管内皮生长因子-碱性成纤维细胞生长因子基因治疗家兔肢体缺血的研究

Gene therapy of vascular endothelial growth factor-basic fibroblast growth factor for hindlimb ischemia in a rabbit model

目的 了解血管内皮生长因子 (VEGF)和碱性成纤维细胞生长因子 (bFGF)联合克隆基因 (VEGF bFGF)治疗兔下肢动脉缺血模型后新生血管和侧支形成状况。方法 应用 40只家兔制成下肢缺血模型 ,其中VEGF bFGF组 10只 ,VEGF组 12只 ,空载体组 8只 ,生理盐水 (NS)组 10只。构建pcDNA3 /VEGF和pcDNA3 /VEGF bFGF真核表达载体。转染缺血部位肌组织 ,行下肢血管造影。结果 血管造影计数显示 ,VEGF bFGF组在转染后 14d(1.98± 0 .2 2 ) ,2 8d (1.81± 0 .5 2 ) ,5 6d(2 .2 1± 0 .44 )和 3个月 (2 .10± 0 .2 2 ) ;VEGF组在 2 8d(1.3 8± 0 .2 9) ,5 6d(1.94± 0 .2 5 )和 3个月 (2 .2 4± 0 .3 1) ,新生血管形成较对照组显著增加 (P <0 .0 5 )。结论 VEGF bFGF真核表达载体可以获得局部高效表达 ,刺激新生血管生成 ,建立侧枝循环 ,改善肢体缺血。

ObjectiveTo study the angiogenesis and collateral circulation formation in a rabbit model of hindlimb ischemia after gene therapy of VEGF-bFGF.Methods40 hindlimb ischemia rabbit models were made:VEGF-bFGF group ( n =20);VEGF group ( n =12);pcDNA_3 group ( n =10);saline group.The hVEGF_ 165 cDNA and hVEGF_ 165-bFGF cDNA were cloned into eukaryotic expression vector pcDNA_3.The pcDNA_3/hVEGF_ 165 and pcDNA_3/hVEGF_ 165-bFGF were administered intramuscularly,and angiography was performed.ResultsThe angiography counting was performed 14 days (1.98± 0.22),28 days (1.81±0.52),56 days (2.21±0.44) and 3 months (2.10±0.22) after the initiation of therapy in VEGF-bFGF treated animals,and 28 days (1.38±0.29),56 days (1.94±0.25) and 3 months (2.24±0.31) after the initiation of therapy in VEGF treated animals.Neovessels and collateral artery development were increased significantly in VEGF-bFGF group or VEGF group as compared with controls ( P <0.05).ConclusionDNA of VEGF-bFGF can be expressed efficiently and locally,stimulate angiogenesis and develop collateral circulation in vivo,and improve hindlimb ischemia.