不同剂量HIF-1α基因对兔缺血后肢血管生成作用的影响  

Effect of different doses of recombinant adenovirus mediated hypoxia-inducible factor-1α on angiogenesis in ischemic lower hind limb of rabbits

作　　者：李明琰[1] 陈建威[1] 张新建[2] 陈冬冬[1] 裴静娴[1] 王月刚[1] 吴平生[1] 

机构地区：[1]南方医科大学南方医院心内科,广东广州510515 [2]中国人民解放军159中心医院麻醉科,河南驻马店463200

出　　处：《南方医科大学学报》2010年第2期210-213,共4页Journal of Southern Medical University

基　　金：国家自然科学基金(30370587)

摘　　要：目的研究不同剂量腺病毒介导的低氧诱导因子-1α(HIF-1α)基因对兔缺血后肢血管新生的影响。方法45只新西兰大白兔建立急性左后肢缺血模型,随机分成5组,每组9只,分别在术后即刻肌肉注射生理盐水0.5ml(NS组)、腺病毒空载体2×1010 PFU(Ad-null组)、腺病毒介导的HIF-1α2×109 PFU(低剂量组)、2×1010 PFU(中剂量组)、2×1011 PFU(高剂量组)。术后第7天行实时PCR检测骨骼肌中HIF-1α mRNA表达水平,术后第0、14、28天对兔后肢进行对比超声检查,术后第28天行病理切片CD31免疫组化微血管密度计数。结果实时PCR显示NS组和Ad-null组HIF-1α基因在mRNA水平表达量相当(P>0.05);各基因转染组HIF-1α表达量高于NS组和Ad-null组,且随转染剂量增加而增加,组间比较差异有统计学意义(P<0.01);对比超声示NS组和Ad-null组A×β标化值无明显差别(P>0.05)。HIF-1α各转染组A×β标化值随剂量增加而增大(高剂量组与NS组、Ad-null组比较,P<0.01;其余各组间比较,P<0.05);微血管密度值各组间比较亦呈上述规律。结论腺病毒介导的HIF-1α基因能促进急性缺血肢体的血管生成,其效果随剂量增加而增大。Objective To investigate the effect of recombinant adenovirus-mediated hypoxia-inducible factor-1α（Ad-HIF-1α） at different doses on angiogenesis in a rabbit model of hind limb ischemia.Methods Left hind limb ischemia was induced in 45 Zealand white rabbits by ligation of the left femoral artery.The rabbits were randomly divided into 5 groups（n=9） to receive intramuscular injections of 0.5 ml saline,2×1010 PFU empty vector（Ad-null）,or different doses of Ad-HIF-1α（2×10^9,2×10^10or 2×10^11 PFU） immediately after the operation.On the 7th day after the operation,real-time PCR was used to detect the expression of HIF-1α mRNA in the skeletal muscles.Immediately and on the 14th and 28th days after the operation,contrast enhanced ultrasound（CEU） was used to observe the blood perfusion of the hind limb.On the 28th day postoperatively,immunohistochemistry for CD31 was performed to evaluate the microvascular density（MVD）.Results Real-time PCR showed that Ad-HIF-1α significantly increased the expression of HIF-1α mRNA（P〈0.01） in a dose-dependent manner as compared with that in the saline and Ad-null groups（P〈0.01）.CEU revealed greater blood perfusion in the hind limb of rabbits in association with increased dose of Ad-HIF-1α（P〈0.05 or P〈0.01）;similar changes in the MVD was observed following Ad-HIF-1α injections as shown by immunohistochemistry（P〈0.05 or P〈0.01）.No significant differences were found either in the blood perfusion or MVD between saline and Ad-null groups（P〉0.05）.Conclusion Ad-HIF-1α can dose-dependently promote the angiogenesis in the ischemic limb of rabbits.

关 键 词：低氧诱导因子-1Α 腺病毒 缺血 后肢 血管生成 实时PCR 

分 类 号：R543[医药卫生—心血管疾病]