关节滑液富集的CC趋化因子配体18通过促进类风湿关节炎成纤维样滑膜细胞迁移参与关节破坏  

Promotion of joint destruction and migration of fibroblast-like synoviocytes subsequent from CC chemokine ligand 18 enriched in synovial fluid in rheumatoid arthritis

作　　者：陈乐锋[1] 荆俊 杨莉娟[1] 马剑达[1] 莫颖倩[1] Chen Lefeng;Jing Jun;Yang Lijuan;Ma Jianda;Mo Yingqian(Department of Rheumatology,Sun Yat-Sen Memorial Hospital,Sun Yat-Sen University,Guangzhou 510120,Guangdong,China)

机构地区：[1]中山大学孙逸仙纪念医院风湿免疫科,广东广州510120

出　　处：《中国医学前沿杂志（电子版）》2023年第11期55-64,共10页Chinese Journal of the Frontiers of Medical Science(Electronic Version)

基　　金：国家自然科学基金青年项目(81601427);广东省基础与应用基础研究基金(2016A030313307,2017A030313470);广东省医学科学技术研究基金(A2017093);中山大学高校基本科研业务费-青年教师培育项目(20ykpy103)。

摘　　要：目的 评估类风湿关节炎(rheumatoid arthritis,RA)关节滑液富集的CC趋化因子配体18(CC chemokine ligand 18,CCL18)与关节破坏的相关性及其可能机制。方法 纳入2018年1月至2023年4月于中山大学孙逸仙纪念医院风湿免疫科就诊的处于疾病活动期的RA患者并随访1年,比较有或无放射学进展者基线关节滑液CCL18水平的差异。体外细胞实验明确关节滑液富集的CCL18或重组CCL18对RA-成纤维样滑膜细胞(fibroblast-like synoviocytes,FLS)或健康FLS(healthy FLS,HFLS)迁移的影响。结果 RA滑液CCL18显著高于配对的血清[差值为(806±609)ng/ml,P<0.001],有1年放射学进展者基线关节滑液CCL18水平显著高于无放射学进展者[(914±166)ng/ml比(264±72)ng/ml,P<0.001]。RA患者滑膜组织多重免疫荧光染色及公开的单细胞测序平台示滑膜CCL18主要由滑膜巨噬细胞产生。使用RA患者滑液预处理24h可激活RA-FLS的迁移活性,而CCL18中和抗体可使RA-FLS迁移率降低40%(P<0.001)。重组CCL18既直接激活RA-FLS或HFLS的迁移活性,又可在Transwell下室发挥趋化作用,两种作用可叠加。采用Oris动态迁移实验连续观察60h,CCL18激活RA-FLS迁移活性呈剂量和时间依赖性。进一步行高通量测序示重组CCL18诱导HFLS产生与RA-FLS相似的基因转录谱。实时荧光定量聚合酶链反应验证,CCL18干预的HFLS中ANXA2、THBS1、TAGLN、NNMT、CDH11、HSPB1、TRIM8、SUMO1等8个与迁移相关基因表达上调。结论 RA滑液富集的CCL18主要由滑膜巨噬细胞产生并通过促进RA-FLS迁移参与关节破坏。Objective To evaluate the correlation between CC chemokine ligand 18(CCL18)enriched in synovial fluid and joint destruction in rheumatoid arthritis(RA)and its possible mechanism.Methods From January 2018 to April 2023,in Department of Rheumatology of Sun Yat-sen University,Sun Yat-sen Memorial Hospital,patients with active RA were included and followed up for 1 year to evaluate radiological progression,and their CCL18 level in serum or synovial fluid was detected at baseline.In vitro cell experiment was used to observe the promotion effect on the migration of RA fibroblast-like synoviocytes(FLS)or healthy FLS(HFLS)by enriched CCL18 or recombinant CCL18 in RA synovial fluid.Results CCL18 in RA synovial fluid was significantly higher than the corresponding serum[(806±609)ng/ml,P<0.001].Baseline CCL18 in synovial fluid was significantly higher in RA patients with 1-year radiographic progression than those without 1-year radiographic progression[(914±166)ng/ml vs.(264±72)ng/ml,P<0.001)].Both immunofluorescence staining of RA synovium and public single-cell sequencing data showed that synovial CCL18 was mainly produced by synovial macrophages.24h pretreatment of RA synovial fluid activated the migration activity of RA-FLS,while neutralizing antibody to CCL18 reduced the mobility of RA-FLS by 40%(P<0.001).Recombinant CCL18 not only directly activated the migration activity of RA-FLS or HFLS,but also played as a chemotactic role in the lower chamber of the Transwell device.Two effects showed superimposed.The dynamic Cell migration for 60h tested by Oris migration assay showed CCL18 promoted the migration of RA-FLS in a dose-dependent and time-dependent manner.Further RNA-sequencing transcriptomics showed that recombinant CCL18 induced HFLS to develop gene expression profiling similar to RA-FLS.Confirmed by real-time quantitative polymerase chain reaction,eight migration-associated genes including ANXA2,THBS1,TAGLN,NNMT,CDH11,HSPB1,TRIM8,and SUMO1 were upregulated in HFLS treated with CCL18.Conclusions CCL18 enriched

关 键 词：类风湿关节炎 成纤维样滑膜细胞 趋化因子配体18 关节破坏 

分 类 号：R593.22[医药卫生—内科学]