Wnt5a participates in hepatic stellate cell activation observed by gene expression profile and functional assays

查看全文 作　　者：Wu-Jun [1]Xiong;Li-Juan [1]Hu;Yi-Cheng [1]Jian;Li-Jing [1]Wang;Ming [1]Jiang;Wei [1]Li;Yi [1]He

机构地区：[1]Department of Gastroenterology,Shanghai East Hospital Affiliated to Tongji University,Shanghai 200120,China

出　　处：《World Journal of Gastroenterology》索引2012年第18卷第15期,共8页

基　　金：Supported by Research Grant for Health Science and Technology of Pudong Health Bureau of Shanghai,No.PKJ2009-Y16 摘　　要：AIM:To identify differentially expressed genes in quiescent and activated hepatic stellate cells(HSCs)and explore their functions.METHODS:HSCs were isolated from the normal Sprague Dawley rats by in suit perfusion of collagenase and pronase and density Nycodenz gradient centrifugation.Total RNA and mRNA of quiescent HSCs,and cultureactivated HSCs were extracted,quantified and reversely transcripted into cDNA.The global gene expression profile was analyzed by microarray with Affymetrix rat genechip.Differentially expressed genes were annotated with Gene Ontology(GO)and analyzed with Kyoto encyclopedia of genes and genomes(KEGG)pathway using the Database for Annotation,Visualization and Integrated Discovery.Microarray data were validated by quantitative real-time polymerase chain reaction(qRTPCR).The function of Wnt5a on human HSCs line LX-2 was assessed with lentivirus-mediated Wnt5a RNAi.The expression of Wnt5a in fibrotic liver of a carbon tetrachloride(CCl4)-induced fibrosis rat model was also analyzed with Western blotting.RESULTS:Of the 28 700 genes represented on this chip,2566 genes displayed at least a 2-fold increase or decrease in expression at a P<0.01 level with a false discovery rate.Of these,1396 genes were upregulated,while 1170 genes were downregulated in culture-activated HSCs.These differentially expressed transcripts were grouped into 545 GO based on biological process GO terms.The most enriched GO terms included response to wounding,wound healing,regulation of cell growth,vasculature development and actin cytoskeleton organization.KEGG pathway analysis revealed that Wnt5a signaling pathway participated in the activation of HSCs.Wnt5a was significantly increased in cultureactivated HSCs as compared with quiescent HSCs.qRTPCR validated the microarray data.Lentivirus-mediated suppression of Wnt5a expression in activated LX-2 resulted in significantly impaired proliferation,downregulated expressions of typeⅠcollagen and transforming growth factor-β1.Wnt5a was upregulated in the fibrotic liver of a CCl4-induced fibrosis rat model.CONCLUSION:Wnt5a is involved in the activation of HSCs,and it may serve as a novel therapeutic target in the treatment of liver fibrosis. 关键词：差异表达基因基因表达谱芯片细胞活化肝星状细胞造血干细胞肌动蛋白细胞骨架聚合酶链反应肝纤维化

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Wnt5a participates in hepatic stellate cell activation observed by gene expression profile and functional assays

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