【摘要】 目的探讨双位点诱变体低氧诱导因子1α(HIF-1α)基因(Ad-H564/803)在动物体内的表达及生物学功能。方法构建急性大鼠下肢缺血模型,随机分为4组,分别以Ad-LacZ、Ad-H0、Ad-H564/803和生理盐水(NS)转染术侧下肢骨骼肌,于转染后1,3,5,7d,RT-PCR测定骨骼肌中HIF-1αmRNA的表达量;基因转染后28d,选择下肢动脉造影及动脉铸型观察血管密度。结果RT-PCR结果显示:Ad-H564/803组HIF-1αmRNA相对表达量较Ad-H0组高(P=0.000);以转染后7d表达量最高(P=0.000);第3,5,7dAd-H564/803组的表达量均高于其它各组(P=0.000)。基因转染28d后造影显示:Ad-H564/803组的侧支血管密度大于Ad-H0组及其他各组;动脉铸型显示:Ad-H564/803组的绒毛样微血管最为密集。结论外源性双位点诱变体HIF-1α基因(Ad-H564/803)可增强急性下肢缺血模型大鼠骨骼肌内HIF-1αmRNA的表达,促进缺血骨骼肌中的血管新生,且生物学功能较野生型HIF-1α基因更强。更多还原

【Abstract】 Objective To study the expression and function of the mutants hyplxia-inducible factor-1α(HIF-1α) in vivo. Methods The acute rat ischemic hindlimb model was established and the Ad-LacZ,Ad-H0,Ad-H564/803 and normal saline(NS)were administered respectively intramuscularly into the ischemic limb. The gene expression was valuated by RT-PCR after 1,3,5,7 days of intramuscular gene transfection in vivo. Selective angiography and vascular cast were performed to illustrate the number of arteries 28th day after intramuscular gene transfection. Results The results of RT-PCR showed that the relative gene expression in group Ad-H564/803(0.6335±0.0331) was higher than that in group Ad-H0 (0.4651±0.0285) in vivo(P = 0.000),and was the highest on 7th day after intramuscular gene transfection(P = 0.000). There was significant difference between the days and the groups after intramuscular gene transfection(P = 0.000). The microvessel density in group Ad-H564/803 was bigger than that in groups Ad-H0,Ad-LacZ and NS. Conclusion These data suggest that gene transfection of the mutants HIF-1α may improve the gene expression of HIF-1α mRNA in muscle and play an important role in modulating angiogenesis in a rat model of hindlimb ischemia. The function of the Ad-H564/803 is more powerful than that of the Ad-H0.更多还原