HIF-1α及VEGF基因促进大鼠缺血组织血管新生的实验研究

Angiogenesis by HIF-1α and VEGF genes in rats with arteriosclerosis obliterans

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 angiogenesis promoting effect of HIF-1α and VEGF genes in rats with arteriosclerosis obliterans. Methods 24 SD rats were randomly divided into 4 groups（naked plasmid group, HIF-1αgroup, VEGF group and HIF-1α＋ VEGF group）. All rats received surgical ligation in left lower limb artery（Arteriosclerosis obliterans animal model）. After surgery, relevant group were respectively intramuscular injection naked plasmid, HIF-1αgene, VEGF gene or HIF-1α＋ VEGF genes. HIF-1α and VEGF were measured by immunohistochemical staining in 7, 14 and 21 days after injection. Microvessel density（MVD） was tested to evaluate eovascularization after 24 days. Result Comparing to naked plasmid group, level of HIF1α was significantly higher in HIF1α group and HIF1α ＋ VEGF group（P 0.05） and level of VEGF was significantly higher in HIF1α group, VEGF group and HIF1α ＋ VEGF group（P〈0.05）. Comparing to HIF1α group, VEGF group and HIF1α ＋ VEGF group, HIF-1α, VEGF and MVD were higher expressed in HIF-1α＋ VEGF group（P〈0.05）.Conclusion HIF-1α＋ VEGF genes can promote angiogenesis in ischaemia.