不同位点诱变体低氧诱导因子1α的动物在体表达比较  

The Gene Expression and the Function of the Mutants HIF-1α in Vivo

作　　者：陈娓[1] 李文强[2] 刘城[1] 韦莉莉[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血管内科,广州510515 [2]新乡医学院第二附属医院中心实验室,河南新乡453002

出　　处：《中国医科大学学报》2009年第3期203-205,共3页Journal of China Medical University

基　　金：国家自然科学基金资助项目(30370587)

摘　　要：目的探讨不同位点诱变体低氧诱导因子1α(HIF-1α)(Ad-H564、Ad-H564/402和Ad-H564/803)转染大鼠急性下肢缺血模型后,骨骼肌中HIF-1α的核酸及蛋白表达。方法建立大鼠急性下肢缺血模型,随机分6组,在骨骼肌中分别注入对照基因Ad-lacZ、野生型HIF-1α基因(Ad-H0)、Ad-H564、Ad-H564/402、Ad-H564/803和生理盐水(NS),于第1,3,5,7d处死动物,RT-PCR检测骨骼肌中HIF-1αmRNA的表达,免疫组化法测7dHIF-1α蛋白表达和28d的毛细血管密度。结果3种突变体HIF-1α基因组的HIF-1αmRNA表达均高于Ad-H0组(F=99.380,P=0.000);以7d的表达量最高(F=24.942,P=0.000);各突变体HIF-1α基因组间相比,以Ad-H564/402组表达量最高,Ad-H564/803组为次,再次为Ad-H564组。各突变体HIF-1α基因组7d的骨骼肌组织间及间质组织中均可见明显的HIF-1α蛋白表达阳性细胞,28d均可见明显的CD31蛋白表达阳性细胞。以Ad-H564/803组最为密集,与对照组差异显著。结论外源性人突变体HIF-1α基因Ad-H564、Ad-H564/402、Ad-H564/803均可促进骨骼肌组织中的HIF-1α核酸及蛋白表达,并促进毛细血管新生,且较Ad-H0的作用更强。各突变体基因组间相比较,Ad-H564/402组HIF-1αmRNA的表达最强,Ad-H564/803组的蛋白表达及促毛细血管新生作用最强。Objective To study the expression and function of the mutants HIF-1α in vivo. Methods The acute rat ischemic hindlimb model were produced and the Ad-LacZ, Ad-H0, Ad-Ad-H564, Ad-H564/402, Ad-H564 and normal saline（ NS ）were administered respectively intramuscularly into the isehemic limb. The gene expression was been evaluated by RT-PCR after 1,3,5,7 days and by immunohistochemistry stain after 7 and 28 days of intramuscular gene transfer in skeletal muscles of rat in vivo. Results The relative gene expression in group of mutants HIF-1α was higher than that in group Ad-H0 in vivo（P = 0.000）,and was highest in the 7th day after intramuscular gene transfer（P =0.000 ）. Them were significant difference between the days and the groups after intramuscular gene transfer （P = 0.000）. The microvessel density in group Ad-H564 was more, than that in group Ad-H0 and as well as Ad-LacZ and NS. Conclusion These data suggest that gene transfer of the mutants HIF-1α may improve the gene expression of HIF-1α mRNA and protein in muscle and play an important vole in modulating angiogenesis in a rat model of hindlimb ischemia. The function of the Ad-H564/402 was power than that of the Ad-H0.

关 键 词：低氧诱导因子1 血管新生 实时荧光定量PCR 免疫组化 

分 类 号：R541.4[医药卫生—心血管疾病]