Gene transfer of angiogenic growth factor for treatment of chronic limb ischemia

以兔后肢缺血模型为模型，研究了腺病毒介导的碱性成纤维细胞生长因子(BFGF)体外基因转移治疗慢性血管闭塞性疾病的新策略。完全切除左股动脉，造成雄性兔后肢缺血状态。同时，从创面切取皮肤切片，培养宿主成纤维细胞。将含有分泌信号序列的改良人bFGF腺病毒载体(AxCAMASFGF腺病毒载体)或含LacZ基因的腺病毒载体(AxCALacZ基因)感染成纤维细胞。股动脉切除后第21天，经左侧髂内动脉注入转基因成纤维细胞。缺血后肢成纤维细胞大量聚集，经AxCAMassbFGF处理后的细胞分泌碱性成纤维细胞生长因子的时间少于14d，而全身碱性成纤维细胞生长因子水平未见升高。接种细胞28d后，测定小牛血压比、血管造影评分、肌肉组织毛细血管密度和左侧髂内动脉血流量，结果显示，移植AxCAMassbFGF细胞的动物较移植AxCALacZ细胞的动物有更多的外侧血管发育。这些结果提示，腺病毒介导的bFGF体外基因转移对改善慢性肢体缺血是有效的。

Adenovirus-mediated ex vivo gene transfer of basic fibroblast growth factor(bFGF), a new strategy for the treatment of chronic vascular occlusive disease, was examined in a rabbit model of hind limb ischemia. The left femoral artery was completely excised to induce an ischemic state in the hind limb of male rabbits. Simultaneously, a skin section was resected from the wound, and host fibroblasts were cultured. The cultured fibroblasts were infected with adenovirus vector containing modified human bFGF cDNA with the secretory signal sequence(AxCAMAssFGF)or LacZ cDNA(AxCALacZ). At 21 days after femoral artery excision, the gene-transduced fibroblasts were administered through the left internal iliac artery. The fibroblasts significantly accumulated in the ischemic hind limb, and the AxCAMAssbFGF-treated cells secreted bFGF for less than 14 days without elevation of systemic bFGF level. At 28 days after cell administration, calf blood pressure ratio, angiographic score, capillary density of muscle tissue and blood flow of the left internal iliac artery were determined, and animals with AxCAMAssbFGF-treated cells showed significantly greater development of cellateral vessels, as compared to those with AxCALacZ-treated cells. These findings suggest that adenovirus-mediated ex vivo gene transfer of bFGF was effective for improvement of chronic limb ischemia.