FAK-ERK信号转导通路在FRNK抑制肝星状细胞胶原合成中的作用  

Role of FAK-ERK signal transduction pathway in hepatic stellate cells collagen synthesis inhibited by FRNK plasmid transfection

作　　者：申建刚 张晓岚[2] 魏娟[2] 霍晓霞[2] 桑荣霞[3] 安君艳[2] 

机构地区：[1]深圳市人民医院,广东省深圳市518003 [2]河北医科大学第二医院消化科,河北省石家庄市050000 [3]石家庄市人民医院消化科,河北省石家庄市050000

出　　处：《世界华人消化杂志》2009年第14期1402-1405,共4页World Chinese Journal of Digestology

基　　金：河北省自然科学基金资助项目; No.C2008001133;河北省卫生厅基金资助项目; No.2003055~~

摘　　要：目的:探讨FAK-ERK信号转导通路在黏着斑激酶相关非激酶(FAK-related non-kinase,FRNK)质粒转染抑制肝星状细胞(hepatic stellate cells,HSCs)胶原合成中的作用.方法:在体外,以FN诱导HSCs增殖,采用脂质体介导的方法用FRNK表达质粒瞬时转染HSCs,利用3H-Pro掺入技术测定HSCsⅠ型胶原的合成,Western blot及RT-PCR方法检测FRNK、FAK、p-FAK(Tyr397)、ERK蛋白和mRNA的表达.结果:FRNK表达质粒成功转染HSCs,在翻译后水平抑制FAK磷酸化.在FRNK转染HSC48h后胶原合成能力较空质粒组显著下降(498.17±73.20vs748.33±61.30,P<0.01).FRNK抑制FAK磷酸化和在翻译和转录水平抑制ERK1、p-ERK的表达,而FN则促进FAK和ERK1、p-ERK在翻译和转录水平的表达.结论:FRNK可以使HSCs胶原合成能力降低,FAK-ERK信号转导通路可能发挥了负调控作用.AIM： To investigate the effect of FAK-ERK signal transduction pathway in hepatic stellate cells （HSCs） collagen synthesis inhibited by FRNK plasmid transfection in vitro. METHODS： After FN stimulated HSCs, FRNK plasmid mediated by cationic liposorne was transfected into HSCs in vitro. HSCs collagen synthesis capability was examined using 3H-Pro incorporation assay. And the protein expression and mRNA expression of FRNK, FAK, p-FAK （Tyr397） and ERK in HSCs were determined using Western blot and RT-PCR, respectively. RESULTS： The expression of FRNK was enhanced after FRNK had been transiently transfected into HSCs in vitro. Compared with the non-FRNK plasmid group, the collagen synthesis in the FRNK plasmid group was significantly inhibited （498.17 ± 73.20 vs 748.33 ± 61.30, P 〈 0.01）. After exposure of HSCs to FRNK plasmid, the protein and mRNA expression levels of p-FAK, ERK1 and p-ERK were dramatically decreased compared with the non-FRNK plasmid group; on the contrary, compared with the con- trol group, the expression levels of p-FAK, ERK1 and p-ERK in FN group were increased. CONCLUSION： After FRNK were transfected successfully into HSCs using lipofectamine, the collagen synthesis in HSCs is inhibited. FAK- ERK signal transduction pathway may be involved in this process.

关 键 词：肝星状细胞 黏着斑激酶相关非激酶 细胞 外信号调节激酶 胶原 免疫蛋白印迹 

分 类 号：R575.2[医药卫生—消化系统]