Rspo3/Lgr5促进N-钙黏蛋白表达参与小鼠肝损伤

doi:10.3969/j.issn.1006-7795.2024.03.015

首都医科大学学报

Rspo3/Lgr5促进N-钙黏蛋白表达参与小鼠肝损伤

高岳, 岳闻慧, 丁靖茹, 李丽英, 杨乐^*

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

收稿日期:2023-11-30 出版日期:2024-06-13 发布日期:2024-06-13
通讯作者: 杨乐 E-mail:yangle@ccmu.edu.cn
基金资助:
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）。

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

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

Received:2023-11-30 Online:2024-06-13 Published:2024-06-13
Supported by:
This work was supported by National Natural Science Foundation of China (82070623), Beijing Natural Science Foundation (7202007), and Scientific Research Common Program of Beijing Municipal Commission of Education (KM202010025029).

摘要/Abstract

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。

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

Abstract: 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.

Key words: liver injury, N-cadherin, R-spondin3, leucine-rich repeat-containing G protein-coupled receptor 5

中图分类号:

高岳, 岳闻慧, 丁靖茹, 李丽英, 杨乐. Rspo3/Lgr5促进N-钙黏蛋白表达参与小鼠肝损伤[J]. 首都医科大学学报, doi: 10.3969/j.issn.1006-7795.2024.03.015.

Gao Yue, Yue Wenhui, Ding Jingru, Li Liying, Yang Le. Rspo3/Lgr5 promotes the expression of N-cadherin and participates in mouse liver injury[J]. Journal of Capital Medical University, doi: 10.3969/j.issn.1006-7795.2024.03.015.

参考文献

［1］程浩, 潘静, 姚甜甜, 等. 全身炎症反应导致肝损伤的临床特点和机制研究现状［J］. 国际病毒学杂志, 2019, 26(5): 354－357.
［2］齐婧姝, 胡旭东, 刘成海. 肝纤维化的逆转机制［J］. 中华肝脏病杂志, 2022, 30(6): 577－582.
［3］ Vanheule E, Geerts A M, Van Huysse J, et al. An intravital microscopic study of the hepatic microcirculation in cirrhotic mice models: relationship between fibrosis and angiogenesis［J］. Int J Exp Pathol, 2008, 89(6): 419－432.
［4］ Yang L, Yue S, Yang L, et al. Sphingosine kinase/sphingosine 1-phosphate (S1P)/S1P receptor axis is involved in liver fibrosis-associated angiogenesis［J］. J Hepatol, 2013, 59(1): 114－123.
［5］ Gerhardt H, Betsholtz C. Endothelial-pericyte interactions in angiogenesis［J］. Cell Tissue Res, 2003, 314(1): 15－23.
［6］ Grant R I, Hartmann D A, Underly R G, et al. Organizational hierarchy and structural diversity of microvascular pericytes in adult mouse cortex［J］. J Cereb Blood Flow Metab, 2019, 39(3): 411－425.
［7］ Hellerbrand C. Hepatic stellate cells—the pericytes in the liver［J］. Pflugers Arch, 2013, 465(6): 775－778.
［8］ Xu J, Gong T, Heng B C, et al. A systematic review: differentiation of stem cells into functional pericytes［J］. FASEB J, 2017, 31(5): 1775－1786.
［9］ Yang L, Dong C B, Yang J J, et al. MicroRNA-26b-5p inhibits mouse liver fibrogenesis and angiogenesis by targeting PDGF Receptor-Beta［J］. Mol Ther Nucleic Acids, 2019, 16: 206－217.
［10］ Crisan M, Yap S, Casteilla L, et al. A perivascular origin for mesenchymal stem cells in multiple human organs［J］. Cell Stem Cell, 2008, 3(3): 301－313.
［11］ Marson R F, Regner A P, Meirelles L D A S. Mesenchymal “stem” cells, or facilitators for the development of regenerative macrophages? Pericytes at the interface of wound healing［J］. Front Cell Dev Biol, 2023, 11: 1148121.
［12］ Globig P, Madurawala R, Römer W R, et al. Mg-based materials diminish tumor spreading and cancer metastases［J］. Bioact Mater, 2023, 19: 594－610.
［13］ Nadeem T, Bogue W, Bigit B, et al. Deficiency of notch signaling in pericytes results in arteriovenous malformations［J］. JCI Insight, 2020, 5(21): e125940.
［14］ Besnier M, Shantikumar S, Anwar M, et al. miR-15a/－16 inhibit angiogenesis by targeting the tie2 coding sequence: therapeutic potential of a miR-15a/16 decoy system in limb ischemia［J］. Mol Ther Nucleic Acids, 2019, 17: 49－62.
［15］ Armulik A, Abramsson A, Betsholtz C. Endothelial/pericyte interactions［J］. Circ Res, 2005, 97(6): 512－523.
［16］ Amsellem V, Dryden N H, Martinelli R, et al. ICAM-2 regulates vascular permeability and N-cadherin localization through ezrin-radixin-moesin (ERM) proteins and Rac-1 signalling［J］. Cell Commun Signal, 2014, 12: 12.
［17］ Steege T E J, Bakker E R M. The role of R-spondin proteins in cancer biology［J］. Oncogene, 2021, 40(47): 6469－6478.
［18］ Nagano K N H. R-spondin signaling as a pivotal regulator of tissue development and homeostasis［J］. Jpn Dent Sci Rev, 2019, 55(1): 80－87.
［19］ Mesci A, Lucien F, Huang X Y, et al. RSPO3 is a prognostic biomarker and mediator of invasiveness in prostate cancer［J］. J Transl Med, 2019, 17(1): 125.
［20］ Scholz B, Korn C, Wojtarowicz J, et al. Endothelial RSPO3 controls vascular stability and pruning through non-canonical WNT/Ca(2+)/NFAT signaling［J］. Dev Cell, 2016, 36(1): 79－93.
［21］ Sweeney M, Foldes G. It takes two: endothelial-perivascular cell cross-talk in vascular development and disease［J］. Front Cardiovasc Med, 2018, 5: 154.
［22］ Gerhardt H, Wolburg H, Redies C. N-cadherin mediates pericytic-endothelial interaction during brain angiogenesis in the chicken［J］. Dev Dyn, 2000, 218(3): 472－479.
［23］ Werner A C, Weckbach L T, Salvermoser M, et al. coronin 1B controls endothelial actin dynamics at cell-cell junctions and is required for endothelial network assembly［J］. Front Cell Dev Biol, 2020, 8: 708.
［24］ Kruse K, Lee Q S, Sun Y, et al. N-cadherin signaling via trio assembles adherens junctions to restrict endothelial permeability［J］. J Cell Biol, 2019, 218(1): 299－316.
［25］ Bonney S K, Coelho-Santos V, Huang S F, et al. Public volume electron microscopy data: an essential resource to study the brain microvasculature［J］. Front Cell Dev Biol, 2022, 10: 849469.
［26］ Jin Y R, Han X H, Nishimori K, et al. Canonical WNT/β-catenin signaling activated by WNT9b and RSPO2 cooperation regulates facial morphogenesis in mice［J］. Front Cell Dev Biol, 2020, 8: 264.
［27］ Annunziato S, Sun T A, Tchorz J S. The RSPO-LGR4/5-ZNRF3/RNF43 module in liver homeostasis, regeneration, and disease［J］. Hepatology, 2022, 76(3): 888－899.
［28］王瑞峰, 张昊驹, 戴宜武, 等. R-脊椎蛋白3对小鼠神经干细胞增殖及Wnt/β-catenin通路的影响［J］. 中华神经医学杂志, 2018, 17(4): 344－348.
［29］ Wootten D, Christopoulos A, Marti-Solano M, et al. Mechanisms of signalling and biased agonism in G protein-coupled receptors［J］. Nat Rev Mol Cell Biol, 2018, 19(10): 638－653.

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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

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