干扰RNA片段对过氧化物酶体增殖物激活受体γ及内皮素-1基因表达的影响

目的:研究干扰RNA片段对过氧化物酶体增殖物激活受体γ(PPARγ)基因表达的影响,了解PPARγ基因表达量发生变化的时候,内皮素-1(ET-1)的基因表达量有无变化,以探讨PPARγ及ET-1基因在动脉粥样硬化致病机制中的作用。方法:采用siRNA Expression Cassette方法干预体外培养的人正常脐静脉内皮细胞,通过RT-PCR、适时定量PCR(quantitative real-time PCR)及Western blot法检测PPARγ及ET-1的mRNA、蛋白质表达水平。结果:针对PPARγ基因设计的siRNA Expression Cassette 1(1 326)使PPARγmRNA及蛋白表达量下调,ET-1 mRNA表达量增多,二者之间表达量的mRNA灰度比值变化有相关性(r=0.995,P=0.042)。siRNAExpression Cassette 2,3,4片断未能发挥干扰效应。结论:干扰RNA片段siRNA Expression Cassette 1(1 326)对PPARγ的基因表达有抑制作用,可以在转录水平抑制PPARγ在人脐静脉内皮细胞的基因表达,同时使ET-1基因表达上调。

Effects of siRNA Segments on the Expression of Peroxisome Proliferator-activated Receptor Gamma and Endothelin-1

Objective: To investigate the influence of siRNA(short interfering RNA) to the expression of PPARγ,to understand the change of endothelin-(ET-1) gene expression when PPARγ diversified,and to explore the role of PPARγ in the mechanism of atherosclerosis.Methods: The HUVEC was intervened with siRNA Expression Cassette segments in vitro.After 24 hours,the mRNA of PPARγ and ET-1 were detected with RT-PCR and quantitative real-time PCR,and the protein of PPARγ was measured with Western blot.Results: siRNA Expression Cassette 1(1 326) down-regulated the mRNA and protein espression of PPARγ and up-regulated ET-1 gene mRNA expression at the same time.A correlation was found between the two genes(r=0.995,P=0.042).Conclusion: siRNA Expression Cassette 1(1 326) can cause an inhibition of PPARγ expression which leads to an increase of ET-1 expression.Therefore,both of PPARγ and ET-1 play a key role in formation of the atherosclerosis.