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目的:观察缺氧诱导因子-1α(HIF-1α)及血管内皮生长因子(VEGF)基因在促进大鼠缺血后肢体血管新生水平。方法:24只大鼠随机分成4组,分别为空载质粒组、HIF-1α组、VEGF组、HIF-1α+VEGF组,每组6只,其中空载质粒组为对照组,HIF-1α组、VEGF组和HIF-1α+VEGF组为实验组。各组均用外科结扎左侧股动脉的方法建立下肢动脉闭塞大鼠模型(ASO大鼠模型)。造模成功后立即每只大鼠缺血局部分别注射空载质粒、HIF-1α裸质粒、VEGF裸质粒剂HIF-1α裸质粒+VEGF裸质粒。分别于7 d、14 d、21 d,3个时间点,各取肌肉样本行HIF-1α、VEGF免疫组织化学检测。术后24 d取标本行估算毛细血管密度,以评价血管新生情况。结果:HIF-1α组和HIF-1α+VEGF组中HIF-1α表达量较对照组明显增高,差异有统计学意义(P<0.05)。HIF-1α组、VEGF组及HIF-1α+VEGF组中VEGF表达较对照组明显增高,差异有统计学意义(P<0.05)。而在各实验组中,HIF-1α+VEGF组中组织新生血管密度增高最为显著。结论:HIF-1α+VEGF基因能促进缺血组织新生血管形成。
Objective: To observe the effects of hypoxia inducible factor-1α (HIF-1α) and vascular endothelial growth factor (VEGF) gene on angiogenesis in rats following ischemia. Methods: Twenty-four rats were randomly divided into 4 groups: empty plasmid group, HIF-1α group, VEGF group and HIF-1α + VEGF group, with 6 rats in each group. Group, VEGF group and HIF-1α + VEGF group as experimental group. All groups were established by surgical ligation of the left femoral artery in the lower extremity arterial occlusion rat model (ASO rat model). Immediately after the success of modeling, each rat was injected with empty plasmid, HIF-1α naked plasmid and naked naked HIF-1α plasmid + VEGF naked plasmid into each rat. HIF-1α and VEGF immunohistochemistry were performed on each of the muscle samples at 7 d, 14 d, 21 d and 3 time points respectively. Twenty-four days after surgery, samples were taken to estimate capillary density to evaluate angiogenesis. Results: The expression of HIF-1α in HIF-1α group and HIF-1α + VEGF group was significantly higher than that in control group (P <0.05). The expression of VEGF in HIF-1α group, VEGF group and HIF-1α + VEGF group was significantly higher than that in control group (P <0.05). In each experimental group, the increase of tissue neovascularization density was the most significant in HIF-1α + VEGF group. Conclusion: HIF-1α + VEGF gene can promote neovascularization in ischemic tissue.