Rspo3/Lgr5促进N-钙黏蛋白表达参与小鼠肝损伤  

Rspo3/Lgr5 promotes the expression of N-cadherin and participates in mouse liver injury

作　　者：高岳 岳闻慧 丁靖茹 李丽英[1] 杨乐 Gao Yue;Yue Wenhui;Ding Jingru;Li Liying;Yang Le(Department of Cell Biology,Municipal Laboratory for Liver Protection and Regulation of Regeneration,Capital Medical University,Beijing 100069,China)

机构地区：[1]首都医科大学细胞生物学系‘肝脏保护与再生调节’北京市重点实验室,北京100069

出　　处：《首都医科大学学报》2024年第3期472-480,共9页Journal of Capital Medical University

基　　金：国家自然科学基金项目(82070623);北京市自然科学基金项目(7202007);北京市教委科技计划一般项目(KM202010025029)。

摘　　要：目的本文旨在研究在损伤肝组织中,特异性顶部盘状底板反应蛋白3(R-spondin3,Rspo3)上调周细胞N-钙黏蛋白(N-cadherin,Ncad,基因名Cdh2)表达,从而参与小鼠肝损伤。方法采用反转录实时定量聚合酶链反应法(reverse transcription-quantitative polymerase chain reaction,RT-qPCR)测定Rspo3,小鼠肝组织富含亮氨酸重复序列G蛋白偶联受体5(leucine-rich repeat-containing G protein-coupled receptor 5,Lgr5)和Cdh2 mRNA表达情况;采用蛋白质免疫印迹方法以及免疫荧光染色方法测定Ncad定位以及表达情况;分离并培养小鼠原代骨髓间充质细胞(bone marrow mesenchymal stromal cell,BMSC),采用靶向Lgr5的RNA干扰实验以确定Rspo3是否通过作用于Lgr5上调Ncad表达。结果在损伤小鼠肝组织中,Rspo3及其受体Lgr5显著上调;肝损伤时黏附连接蛋白Ncad表达上调,且与Rspo3及Lgr5表达量呈正相关;Ncad主要定位于损伤肝脏的周细胞;使用Rspo3处理小鼠原代BMSC能够上调Ncad表达,Lgr5 siRNA能使Ncad水平下降。结论在小鼠损伤肝组织中,Rspo3通过作用于受体Lgr5,上调周细胞中Ncad表达,从而影响小鼠肝损伤。Objective The aim of this study is to investigate R-spondin3(Rspo3)up-regulates the expression of Ncad(N-cadherin,gene named Cdh2)and participates in mouse liver injury.Methods The expression of Rspo3,leucine-rich repeat-containing G protein-coupled receptor 5(Lgr5)and Cdh2 were measured by reverse transcription-quantitative polymerase chain reaction(RT-qPCR).The expression of Ncad was measured by Western blotting and Immunofluorescence.Serum-starved primary cultured mouse bone marrow mesenchymal stromal cell(BMSC)were transfected with Lgr5 siRNA to verify whether Rspo3 up-regulates Ncad expression by acting on Lgr5.Results The Rspo3 and Lgr5 mRNA expression was up-regulated in the liver tissue of mouse injured liver model.The mRNA expression of Cdh2 was up-regulated and positively correlated with the expression of Rspo3 and Lgr5,which is one of the receptors of Rspo3.Immunofluorescence shows that Ncad is mainly localized in the pericytes of the mouse injured liver.The Rspo3-induced up-regulation of Ncad was inhibited after Lgr5 siRNA transfection.Conclusion In the mouse injured liver tissue,Rspo3 up-regulates the expression of Ncad in pericytes by acting on Lgr5,and affects mouse liver injury.

关 键 词：肝损伤 N-钙黏蛋白 特异性顶部盘状底板反应蛋白3 富含亮氨酸重复序列G蛋白偶联受体5 

分 类 号：R34[医药卫生—基础医学]