TGF-β1通过ERK12信号通路促进人肝星状细胞LX-2中HMGA1基因的表达.pdfVIP

Abstract ABSTRACT Objective Theaimofthis stud istoinvestigatewhetherTGF-β1promotesthe expression ofHMGA1throughERK1/2andorAktpathway. Methods In order to elucidate the molecular mechanism whereb TGF-β1up-regulates HMGA1 ,we incubated the LX-2 cells with different concentration of TGF-β1,and detectedthe mRNA level ofHMGA1 with Realtime-qPCR.Further,weused TGF-β1 and PD98059 (MK2206) to treat LX-2 cells,and used western blot to detect the protein level of pERK1/2,pAkt,ERK1/2,Akt and HMGA1, used Realtime-qPCR to detectthemRNAlevelofHMGA1. Results 1 We observed a dose-dependentl increase of HMGA1 in the presence of 、 TGF-β1,whichwasuptoapeakat5ng/ml. 2 WeshowedthatTGF-β1markedl inducedtheactivationofERK1/2andAkt 、 pathway. 3、WeshowedthatTGF-β1markedl inducedtheexpressionofHMGA1，andthe inhibitorofERK1/2pathwa significantl blockedtheeffectsofTGF-β1 whilethe ， inhibitorofAktpathwa hadnoeffects. Conclusions 1 TGF-β1markedl inducestheexpressionofHMGA1inLX-2HSCcells. 、 2 TGF-β1markedl inducestheactivationofERK1/2andAktpathwa inLX-2 、 HSCcells. 3 TGF-β1promotestheexpressionofHMGA1throughERK1/2pathway. 、 KeyWords TGF-β1 HMGA1 ERK1/2 Akt LX-2HepaticStellateCells ； ； ； ； ； LiverFibrosis IV 目录 目 录 1 1 第 章 前言 2 4 第 章 材料与方法 2.1 实验材料4 2.1.1 细胞株4 2.1.2 主要试剂和材料4 2.1.3 主要仪器设备5 2.2 实验方法6 2.2.1 主要试剂的配制方法6 2.2.2 人肝星状细胞株LX-2 的培育8 2.2.3TGF-β1 HMGA1 HMGA2 8 浓度梯度对 及 表达的影响