抑制内质网应激关键信号通路蛋白eIF2α去磷酸化与肝细胞增殖及c-Met表达的关系  

Relationships of inhibiting dephosphorylation of eIF2α,a key signaling pathway for ERS,with hepatocyte proliferation and c-Met expression

作　　者：陈欢 蒋小铃 李吉贵 方家琴 冯素敏 何毅怀 陈娅 CHEN Huan;JIANG Xiaoling;LI Jigui;FANG Jiaqing;FENG Sumin;HE Yihuai;CHEN Ya(Department of Internal Medicine,The Fifth People's Hospital of Zunyi,Zunyi 563000,China;不详)

机构地区：[1]遵义市第五人民医院内科,遵义563000 [2]遵义医科大学附属医院感染科

出　　处：《山东医药》2023年第2期20-23,共4页Shandong Medical Journal

基　　金：国家自然科学基金项目(81560110);贵州省卫生健康委科学技术基金项目(gzwjkj2021-071,gzwjkj2020-1-041);贵州省科技计划项目[黔科合支撑(2019)2803号];贵州省遵义市科技计划项目[遵市科合HZ字(2021)58号];遵义医科大学学术新苗培养及创新探索专项项目[黔科合平台人才(2017)5733-013]。

摘　　要：目的探讨抑制内质网应激(ERS)关键信号通路蛋白真核细胞起始因子2α(eIF2α)去磷酸化与肝细胞增殖及促肝细胞生长因子受体(c-Met)表达的关系。方法将人肝细胞株L02分为对照组和胡萝卜素(TG)组,TG组加入含内质网钙平衡抑制剂毒胡萝卜素(TG)的RPMI1640培养基,诱导构建细胞ERS模型;对照组仅加入RPMI1640培养基,培养48 h。采用CCK-8检测细胞活力,Western blotting法检测细胞ESR相关蛋白p-eIF2α、ERAD通路相关蛋白HRD1、XBP1以及c-Met蛋白表达。另取L02细胞,分为TG组和Salubrinal+TG组,Salubrinal+TG组给予eIF2α去磷酸化抑制剂Salubrinal预处理2 h,然后加入1μmol/L TG培养48 h;TG组加入1μmol/L TG培养48 h。采用CCK-8检测细胞活力,Western blotting法检测细胞eIF2α、p-eIF2α、HRD1、XBP1、c-Met蛋白表达。结果与对照组相比,TG组p-eIF2α、HRD1表达水平升高,TG组ERS相关蛋白p-eIF2α表达上调,ERAD通路激活,表明ERS细胞模型构建成功。TG组细胞增殖活力和c-Met蛋白表达水平均低于对照组(P均<0.05)。与TG组相比,Salubrinal+TG组细胞XBP1、HRD1蛋白表达降低,细胞增殖活力和c-Met蛋白表达升高(P均<0.05)。结论在肝细胞ERS中,抑制eIF2α去磷酸化能够部分恢复肝细胞细胞增殖活力及c-Met表达,促进细胞存活。Objective To investigate the relationships of inhibiting the dephosphorylation of eukaryotic translation initiation factor 2α(eIF2α),a key signaling pathway protein of ERS,with hepatocyte proliferation and hepatocyte growth factor receptor(c-Met)expression.Methods Human liver cell line L02 was divided into the control group and thapsigargin(TG)group.In the TG group,ERS model was induced by adding RPMI1640 medium containing endoplasmic reticulum calcium balance inhibitor TG.In the control group,we only added RPMI1640 medium.At 0,24,48 h of culture,CCK-8 was used to detect cell viability,and Western blotting was used to detect the expression levels of ERS-related proteins p-eIF2α,ERAD path-related proteins HRD1,XBP1,and c-Met.L02 cells were divided into TG group and Salubrinal+TG group.L02 cells in the Salubrinal+TG group were pretreated with eIF2αdephosphorylation inhibitor Salubrinal for 2 h,and then were cultured with 1μmol/L TG for 48 h.L02 cells in the TG group were added with 1μmol/L TG and cultured for 48 h.CCK-8 was used to detect cell viability,and Western blotting was used to detect protein expression levels of eIF2α,p-eIF2α,HRD1,XBP1,and c-Met.Results Compared with the control group,the expression levels of peIF2αand HRD1 in TG group increased,the expression of ERS-related protein p-eIF2αin TG group was up-regulated,and the ERAD pathway was activated,indicating that the ERS cell model was successfully constructed.The cell proliferation activity and c-Met protein expression levels in the TG group were lower than those in the control group(all P<0.05).Compared with TG group,the protein expression levels of XBP1 and HRD1 decreased,while the proliferative activity and c-Met protein expression increased in the Salubrinal+TG group(all P<0.05).Conclusion Inhibition of eIF2αdephosphorylation can partially restore hepatocyte proliferation and c-Met expression in ERS,and promote cell survival.

关 键 词：内质网应激 促肝细胞生长因子受体 肝细胞增殖 真核细胞起始因子2α 肝损伤 

分 类 号：R575[医药卫生—消化系统]