缺氧诱导因子-1α小片段干扰性RNA对人血管内皮细胞缺氧诱导因子-1α表达的调控  

Regulation of HIF-1α expression by small interfering RNA targeting HIF-1α in human vascular endothelial cells

作　　者：蒋剑[1] 夏晓波[1] 许惠卓[1] 熊思齐[1] 刘双珍[1] 

机构地区：[1]中南大学湘雅医院眼科,湖南长沙410008

出　　处：《眼视光学杂志》2007年第6期377-379,共3页Chinese Journal of Optometry & Ophthalmology

基　　金：国家自然科学基金资助项目(030471853)。

摘　　要：目的探讨缺氧诱导因子-1α(hypoxia inducible factor-1α,HIF-1α)小片段干扰性RNA(siRNA)对人血管内皮细胞HIF-1α表达的影响。方法构建HIF-1αsiRNA重组质粒。将体外培养的人血管内皮细胞(HUVEC-12)分成常氧(20%)组和低氧(1%)组。低氧组又分为对照组、空载体组和HIF-1α组。采用脂质体LipofectamineTM 2000分别转染空载体质粒(空载体组)、HIF-1αsiRNA(HIF-1α组),对照组不作转染。采用SP免疫细胞化学法检测各组细胞中HIF-1α蛋白表达的变化,并进行计算机图像分析。结果对照组和空载体组细胞HIF-1α蛋白表达较常氧组增强,而HIF-1α组较对照组明显减弱。结论HIF-1αsiRNA能显著抑制HIF-1α的表达,为视网膜新生血管的基因治疗提供了新方法。Objective To investigate the effect of small interfering RNA （siRNA） targeting hypoxia inducible factor-1α （HIF-1α） on the expression of HIF-1α in human vascular endothelial cells. Methods HIF-1α siRNA recombinant plasmid was constructed. Human vascular endothelial cells were cultured in vitro and divided into a normoxia group （20%） and a hypoxia group （1%）. The hypoxia group was divided into a control group, vector group and HIF- 1α group, Liposome mediated HIF-1α siRNA complex was transfected to the cells in the HIF-1α group, Liposome with vector plasmid complex was transfected to the cells in the vector group. No transfection was done in the control group. The expression of the HIF-1α protein was tested with immunocytochemical staining （SP method）. The results were analyzed by a computer image-analysis system. Results There was a strong expression of HIF-1α protein in the control and vector groups compared to the nonnoxia group. The expression of HIF-1α protein in the HIF-1α group decreased significantly compared to the control group. Conclusion HIF-1α siRNA effectively inhibits the expression of HIF-1α and provides a new approach to gene therapy in retinal neovascularization.

关 键 词：缺氧诱导因子-1Α RNA干扰 血管内皮细胞 

分 类 号：R774.1[医药卫生—眼科]