β1,4-GalT-I通过调节CXCL10/CXCR3激活影响RA成纤维样滑膜细胞的侵袭  

β1,4-GalT-I affects the fibroblast-like synoviocytes invasion in RA by regulating CXCL10/CXCR3 activation

作　　者：朱新辉[1] 孙驰 徐大伟[1] 崔胜宇[1] 刘巍[1] 王友华[2] 崔志明[1] ZHU Xinhui;SUN Chi;XU Dawei;CUI Shengyu;LIU Wei;WANG Youhua;CUI Zhiming(Department of Orthopaedics,the First People′s Hospital of Nantong City,Jiangsu Province,Nantong 226001;Department of Orthopedics,the Affiliated Hospital of Nantong University)

机构地区：[1]江苏省南通市第一人民医院骨科,南通226001 [2]南通大学附属医院骨科

出　　处：《南通大学学报（医学版）》2020年第2期122-127,共6页Journal of Nantong University(Medical sciences)

基　　金：南通市科技计划项目(JCZ18011,JC2018020,MB2019001)。

摘　　要：目的:探讨β1,4-半乳糖基转移酶-I(β1,4-galactosyltransferase-I,β1,4-GalT-I)与趋化因子CXC配体10(CXC ligand 10,CXCL10)/CXC受体3(CXC receptor 3,CXCR3)在类风湿关节炎(rheumatoid arthritis,RA)中的相互关系及其在RA成纤维样滑膜细胞(fibroblast-like synoviocyte,FLS)侵袭中的作用。方法:(1)建立胶原诱导的关节炎(collagen-induced arthritis,CIA)小鼠模型,评估β1,4-GalT-I的作用;(2)运用免疫共沉淀和免疫荧光双标检测β1,4-GalT-I与CXCR3间的相互作用;(3)运用凝集素结合测定检测Galβ1,4GlcNAc糖链的合成;(4)运用酶联免疫吸附试验(enzyme linked immunosorbent assay,ELISA)检测截短基质金属蛋白酶-1(matrix metalloproteinase-1,MMP-1)水平;(5)运用Transwell分析检测FLS侵袭性的变化。结果:(1)抑制β1,4-GalT-I减轻CIA小鼠模型的炎症反应;(2)β1,4-GalT-I与CXCR3在体外和体内存在相互作用并在FLS中存在共定位;(3)肿瘤坏死因子α(tumor necrosis factor-α,TNF-α)刺激细胞时,CXCR3上的Galβ1,4GlcNAc糖链合成显著增加;(4)在CXCR3转染的细胞中,si-β1,4-GalT-I降低了截短MMP-1的水平,当使用CXCR3抑制剂AMG487时,β1,4-GalT-I不能增加截短MMP-1的表达;(5)β1,4-GalT-I影响CXCL10/CXCR3介导的FLS侵袭性。结论:β1,4-GalT-I通过调节CXCL10/CXCR3激活影响RA成纤维样滑膜细胞的侵袭。Objective:To explore the relationship betweenβ1,4-galactosyltransferase-I(β1,4-GalT-I)and chemokine CXC ligand 10(CXCL10)/CXC receptor 3(CXCR3)in rheumatoid arthritis(RA)and its role in the invasion of RA fibroblast-like synoviocytes(FLS).Methods:(1)Collagen-induced arthritis(CIA)mouse models were established to evaluate the effect ofβ1,4-GalT-I;(2)Immunoprecipitation and double label immunofluorescence were used to detect the interaction betweenβ1,4-GalT-I and CXCR3;(3)Lectin binding assay was used to analyse the synthesis of Galβ1,4GlcNAc sugar chain;(4)Levels of truncated matrix metalloproteinase-1(MMP-1)were detected by enzyme linked immunosorbent assay(ELISA);(5)Transwell analysis was used to detect the invasiveness changes of FLS.Results:(1)Inhibition ofβ1,4-GalT-I reduced inflammation in CIA mouse models;(2)β1,4-GalT-I interacted with CXCR3 in vitro and in vivo,and they co-localized in FLS;(3)When tumor necrosis factor-α(TNF-α)stimulated FLS,the sugar chain of Galβ1,4GlcNAc on CXCR3 increased significantly;(4)In CXCR3 transfected cells,si-β1,4-GalT-I reduced the levels of truncated MMP-1,andβ1,4-GalT-I could not increase the expression of truncated MMP-1 by using the CXCR3 inhibitor AMG487;(5)β1,4-GalT-I affected CXCL10/CXCR3-mediated FLS invasiveness.Conclusion:β1,4-GalT-I may affect FLS invasion in RA by regulating CXCL10/CXCR3 activation.

关 键 词：类风湿关节炎 β1 4-半乳糖基转移酶-I CXC受体3 成纤维样滑膜细胞 CXC配体10 

分 类 号：R593.22[医药卫生—内科学]