siRNA抑制IGFBPrP1表达对肝星状细胞株分泌细胞外基质的影响  

Effects of knocking down IGFBPrP1 with siRNA on extracellular matrix secretion in HSC-T6

作　　者：刘立新[1] 邱志红[1] 张骞骞[1] 

机构地区：[1]山西医科大学第一医院科研实验中心肝病研究所省部共建教育部细胞生理学重点实验室,太原030001

出　　处：《临床肝胆病杂志》2011年第3期248-250,共3页Journal of Clinical Hepatology

基　　金：国家自然科学基金(30871146);教育部新世纪优秀人才支持计划(NCET-06-0264)

摘　　要：目的利用化学合成的siRNA抑制肝星状细胞(HSC)中胰岛素样生长因子结合蛋白相关蛋白1(IGFBPrP1)基因的表达,观察其对HSC产生细胞外基质(ECM)的影响,以明确IGFBPrP1在肝纤维化中的作用地位。方法 (1)化学合成2对针对IGFBPrP1基因的siRNAs,转染肝星状细胞株HSC-T6,筛选抑制率较高的siRNA用于干扰实验;(2)将HSC-T6分为3组:正常对照组、阴性对照组和IGFBPrP1 siRNA干扰组。将筛选的抑制效率较高的siRNA转染HSC-T6,Western Blot检测其IGFBPrP1、Ⅰ型胶原(ColⅠ)和纤维连接蛋白(FN)的表达。采用单因素方差分析,多重比较采用SNK-q检验。结果 (1)将2对IGFBPrP1 siRNA转染HSC,筛选出能高效抑制IGFBPrP1表达的siRNA;(2)与正常对照组和阴性对照组相比,IGFBPrP1 siRNA干扰组能明显抑制IGFBPrP1、ColⅠ和FN的表达(F=48.018、81.246、43.850,P<0.01)。结论 IGFBPrP1 siRNA能特异性抑制HSC中IGFBPrP1的表达,进而使HSC中ECM的重要成分ColⅠ和FN合成和分泌减少,故IGFBPrP1可在肝纤维化中起有重要作用。Objective To investigate the effects of inhibiting insulin-like growth factor binding protein related protein1（IGFBPrP1） by chemically synthesized small interfering RNA（siRNA） on the secretion of extracellular matrix（ECM） in HSC-T6 and to determine the effect of IGFBPrP1 on the hepatic fibrosis.Methods（1）Two pairs of chemically synthesized siRNAs targeting IGFBPrP1 were respectively transfected into HSC-T6 cells for evaluation of silence efficacy and the one with higher efficacy was used in the following experiments;（2）Divide the HSC-T6 cells into three groups as follows： normal control group,negative control group and IGFBPrP1 siRNA group.HSC-T6 was treated by IGFBPrP1 siRNA,then the expressions of IGFBPrP1 and collagenⅠand fibronectin were detected by Western Blot.The data was analyzed by one-way ANOVA and SNK-q test.Results（1）Two pairs of siRNAs were transfected into HSC-T6.One pair of siRNA,which can highly effectively inhibit expression of IGFBPrP1 gene was screened successfully;（2）The expressions of IGFBPrP1 and collagenⅠand fibronectin in IGFBPrP1 siRNA group were significantly decreased than those in normal control group and negative control group（F﹦48.018,81.246,43.850,P﹤0.01）.Conclusion siRNA targeting IGFBPrP1 specifically inhibits the expression of IGFBPrP1 in HSC resulting in decreased synthesis and secretion of collagenⅠand fibronectin in hepatic stellate cells.So IGFBPrP1 can play an important role in liver fibrosis.

关 键 词：RNA 小分子干扰 胰岛素样生长因子结合蛋白质类 肝细胞 细胞外基质 

分 类 号：R575.2[医药卫生—消化系统]