HB-EGF对HSC-T6细胞Ras/ERK通路的激活及对MMP-2、TIMP-1表达的影响  

Effect of HB-EGF on the Activation of Ras/ERK Signal Pathway and the Expression of MMP-2,TIMP-1 in HSC-T6 cells

作　　者：张莉娟[1] 何彬彬[1] 黄月红[1] 陈治新[1] 王小众[1] 

机构地区：[1]福建医科大学附属协和医院消化内科,福州350001

出　　处：《福建医科大学学报》2016年第2期69-73,共5页Journal of Fujian Medical University

基　　金：福建省临床医学重点专科资助项目(闽卫科教[2012]149号)

摘　　要：目的观察肝素结合性表皮生长因子样生长因子(HB-EGF)激活肝星状细胞(HSC)过程中Ras/ERK信号通路的变化和对金属基质蛋白酶(MMP)及其抑制物的影响。方法传代培养HSC-T6细胞,分空白对照组、HB-EGF处理组(20,40,80ng/mL),采用QRT-PCR法检测MMP-2、金属蛋白酶组织抑制剂-1(TIMP-1)及表皮生长因子受体(EGFR)mRNA的表达,并确定HB-EGF刺激HSC-T6Ras/ERK通路(ERK1、ERK2、P38MAPK)活化的最佳浓度及最佳时间,检测该通路活化情况。结果 QRT-PCR检测提示,MMP-2的表达随HB-EGF干预时间的延长增加,TIMP-1的表达随干预时间的延长递减;40ng/mL HB-EGF培养HSC-T6 48h后,EGFR mRNA的表达较空白对照组明显上调(P<0.05);选择40ng/mL HB-EGF培养48h为最佳刺激时间与浓度,该处理组ERK1mRNA的表达较空白对照组明显上调(P<0.05),但ERK2、P38MAPK的变化无统计学意义;Western-blot检测提示该处理组ERK1蛋白的表达较空白对照组上调(P<0.05)。结论HB-EGF可影响HSC-T6的MMP-2、TIMP-1的表达;HB-EGF通过促进HSC-T6表达EGFR、ERK1,有效激活细胞内Ras/ERK信号通路。Objective To investigate the effect of HB-EGF on Ras/ERK signaling transduction pathway and MMPs,TIMPs of HSC. Methods The rat hepatic stellate cells T6（HSC-T6）were treated with DMEM supplemented with 2%FBS,HB-EGF（20,40,80ng/mL）for 24 or 48h. We used QRTPCR to detect the expression of MMP-2and TIMP-1＇s mRNA in cells stimulated by HB-EGF at different concentrations and times. We analyzed EGFR gene at the level of transcription by QRT-PCR,and determined a best concentration and time of HB-EGF（40ng/mL,48h）to stimulate the Ras/ERK signaling transduction pathway（ERK1,ERK2,P38MAPK）in HSC-T6 cells before detecting the activation of Ras/ERK pathway. Results MMP-2 mRNA and protein were increased in time-dependent pattern,while TIMP-1was decreased;EGFR mRNA was increased when cells cultured with HB-EGF 48 hat concentration of 40ng/mL,compared to the control group（P〈0.05）. Compared with control group,the expression of ERK1＇s mRNA and protein were much higher after the treatment with 40ng/mL HB-EGF48h;but no significant differences were found in the ERK2 and P38MAPK mRNA and protein.Conclusion HB-EGF has no appreciable effect on MMP-2and TIMP-1in HSC-T6;HB-EGF can promote EGFR and ERK1＇s expression to activate the Ras/ERK signal transduction pathway in HSC-T6 cells.

关 键 词：表皮生长因子 肝素 肝细胞 星形细胞 大鼠 细胞外信号调节MAP激酶类 RAS蛋白质类 胞间信号肽类和蛋白质类 基质金属蛋白酶类 金属蛋白酶1组织抑制剂 

分 类 号：R575.2[医药卫生—消化系统]