gp73在小鼠肝纤维化肝组织中的表达及机制  

Expression and mechanism of gp73 in liver tissue of mouse liver fibrosis

作　　者：马玲玉 甄一宁 罗云萍[1] 段昭君 MA Ling-yu;ZHEN Yi-ning;LUO Yun-ping;DUAN Zhao-jun(Department of Immunology,Institute of Basic Medical Science CAMS,School of Basic Medicine PUMC,Beijing 100005,China)

机构地区：[1]中国医学科学院基础医学研究所北京协和医学院基础学院免疫学系,北京100005

出　　处：《基础医学与临床》2020年第6期771-776,共6页Basic and Clinical Medicine

基　　金：国家自然科学基金(81601374)。

摘　　要：目的在小鼠肝组织中探讨高尔基体跨膜糖蛋白73(gp73)在肝纤维化进程中的变化及机制。方法腹腔注射CCl4构建C57BL/6J小鼠肝纤维化模型;用ELISA检测小鼠血清中gp73的水平;用免疫组织化学染色检测肝组织内gp73的表达;用密度梯度离心法分离肝星状细胞(HSC)并检测其中gp73的表达;用免疫组织化学染色法检测肝组织中胰岛素样生长因子2 mRNA结合蛋白3(IGF2BP3)的表达;接下来分别转染IGF2BP3过表达与敲低的质粒,用RT-PCR检测GP73蛋白的编码基因GOLM1的表达变化;流式细胞计量术检测基因敲除鼠肝纤维化模型中HSC内gp73的表达。结果在肝纤维化模型的小鼠血清(P<0.01)、肝组织(P<0.05)及原代HSC(P<0.001)中gp73蛋白表达水平均升高,同时组织中的IGF2BP3蛋白表达也升高(P<0.05)。细胞系中转入IGF2BP3的过表达及敲低质粒后,成功过表达IGF2BP3(P<0.001)和敲低(P<0.01),GOLM1的表达也随之上调(P<0.001)和下调(P<0.01)。随后在IGF2BP3敲除鼠的肝纤维化模型的HSC中检测发现gp73表达降低(P<0.01)。结论在CCl4诱导的小鼠肝纤维化模型中,由HSC表达的gp73蛋白水平升高,而RNA结合蛋白IGF2BP3是促进其表达的潜在调控因素。Objective To investigate the changes and mechanism of golgi protein 73(gp73)in mouse with liver fibrosis.Methods The model of hepatic fibrosis was developed by intraperitoneal injection of CCl4 in C57 BL6/J mice.Serum level of gp73 was determined by ELISA.The expression of gp73 in liver tissues was detected by immuno-histochemical staining.Hepatic stellate cell(HSC)was isolated by density gradient centrifugation and gp73 expression was detected.The expression of insulin like growth factor 2 mRNA binding protein 3(IGF2 BP3)in liver tissues was detected by immuno-histochemical staining.Next,IGF2 BP3 over-expression and knock down plasmids were transfected,and the changes of GOLM1,which encodes gp73,were detected by RT-PCR.Furthermore,the expression of gp73 in hepatic stellate cells of fibrotic IGF2BP3 knockout mice was detected by flow cytometry.Results A higere expression of gp73 protein increased in serum(P<0.01),liver(P<0.05)and primary hepatic stellatecells(P<0.001)in the fibrotic mice model was found as compared to the control animals.Along with that,the protein level of IGF2BP3 in the liver was also increased(P<0.05).After over-expressing or knocking down IGF2BP3 by transient transfection,the mRNA expression of GOLM1 was also up-regulated(P<0.001)and down-regulated(P<0.01).Respectively,compared with the wide type group,the expression of gp73 was decreased in the hepatic stellate cells of fibrotic IGF2BP3 conditional knockout mice(P<0.01).Conclusions In the CCl4 induced liver fibrosis mice,the protein level of gp73 protein in hepatic stellate cells is increased,and RNA binding protein IGF2 BP3 is a potential regulatory factor to promote its expression.

关 键 词：肝纤维化 GP73 IGF2BP3 

分 类 号：R735.7[医药卫生—肿瘤]