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

doi:10.3969/j.issn.1006-7795.2024.03.015

Abstract

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

CLC Number:

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.

References

［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.

[1]	. A view of sulforaphane interference with microtubules and mitochondrial kinetics in anti-cancer mechanisms [J]. Journal of Capital Medical University, 2023, 44(3): 363-369.
[2]	Yang Min, Yan Xiang, Liu Xijuan, An Guo, Ding Huirong, Cai Ying, Ge Huizheng. Establishment and characterization of immune cells in an orthotopic cell-derived xenograft mouse model of human lung cancer using multi-color flow cytometry [J]. Journal of Capital Medical University, 2023, 44(1): 99-106.
[3]	Li Peixin, Rao Zhijian, Huang Hu. A study of the effect of ghrelin on hypothalamic agouti-related protein expressing by using GT1-7 cell lines [J]. Journal of Capital Medical University, 0, (): 924-930.
[4]	Ni Mingyue, Liu Yuanli, Tu Zhenzhen, Guo Liyu, Zang Dandan, Zhou Haisheng. Transcription factor GRHL3 regulates the expression of GPR108 gene and its mechanism [J]. Journal of Capital Medical University, 2022, 43(3): 446-455.
[5]	Li Jun, Li Shuoshuo, Peng Zhixin, Liao Yajin, Cheng Jinbo, Yuan Zengqiang. Role of microglial histone deacetylases 3 in hypobaric hypoxia-induced oxidative stress [J]. Journal of Capital Medical University, 2022, 43(1): 91-98.
[6]	Li Binghui. Chemical nature of metabolic reprogramming——Professor Li Binghui [J]. Journal of Capital Medical University, 2020, 41(5): 795-799.
[7]	Gong Mingtao, Zhang Chenguang, Ding Wei. Genetically-encoded calcium indicator for detecting the dynamics of drug-induced nuclear calcium signals in real time [J]. Journal of Capital Medical University, 2018, 39(3): 378-384.
[8]	Yang Jian, Wang Yong, Wang Xuan, Yang Zhaofei, Gong Xiaoli, Wang Xiaomin. Parkinson's disease related gene PINK1 but not DJ-1 was down-regulated in neurotoxicity of 6-hydroxydopamine and 1-methyl-4-phenylpyridinium [J]. Journal of Capital Medical University, 2015, 36(5): 740-746.
[9]	Zhang Jianhua, Dong Chunxia, Ren Fanggang, Tan Yanhong, Yang Linhua. Isolation and culture of human osteoclasts from peripheral blood and the interaction between osteoclasts and myeloma cells [J]. Journal of Capital Medical University, 2014, 35(5): 572-576.
[10]	Zhang Jing, Liu Wei, Chen Yunyu, Si Shuyi. Screening and activity evaluation for potential inhibitors of human PLK1 [J]. Journal of Capital Medical University, 2014, 35(3): 337-343.
[11]	Zhang Ning, Huang Jian. Neurofibromatosis type 2 gene mutation and its expression and tumors of digestive system [J]. Journal of Capital Medical University, 2014, 35(2): 257-260.
[12]	HE Xiujuan, LIN Yan, LV Zhe, AN Yunqing, DING Yuezhong, LI Ping. Effects of Zhuhonggao on the expression of cytokines and adhesion molecule in THP-1 cells induced by lipopolysaccharide [J]. Journal of Capital Medical University, 2013, 34(4): 572-575.
[13]	Ma Liang, Zhao Hailing, Zhao Tingting, Zhao Meimei, Liu Yi, Gao Peng, Li Ping, Cao Yongtong. Effect of EPHX2 rs751141 mutation on renal microinflammation and its molecular mechanism [J]. Journal of Capital Medical University, 2021, 42(5): 691-697.