RA滑膜成纤维样细胞外泌体促进巨噬细胞向M1极化  

M1 polarization is induced by exosomes derived from fibroblast-like synoviocytes from patients with rheumatoid arthritis

作　　者：虞珊珊[1] 李静[1] 徐臻 成宇 宗明[1] 范列英[1] YU Shanshan;LI Jing;XU Zhen;CHENG Yu;ZONG Ming;FAN Lieying(Clinical Laboratory,Shanghai East Hospital,School of Medicine,Tongji University,Shanghai 200120,China)

机构地区：[1]同济大学附属东方医院检验科,上海200120

出　　处：《同济大学学报（医学版）》2023年第6期785-791,共7页Journal of Tongji University(Medical Science)

基　　金：国家自然科学基金面上项目(81971535);上海市浦东新区卫生系统优秀青年医学人才培养计划(PWRq2020-21);上海市“医苑新星”青年医学人才培养资助计划(YYXX202101)。

摘　　要：目的 探讨类风湿关节炎滑膜成纤维样细胞(rheumatoid arthritis fibroblast-like synoviocytes, RA-FLS)外泌体在调控巨噬细胞(macrophage, M■)极化中的作用。方法 收集6例骨关节炎(osteoarthritis, OA)患者和6例类风湿关节炎(rheumatoid arthritis, RA)患者关节液和滑膜组织,流式微珠阵列(CBA)法检测关节液中M1和M2型细胞因子的表达;免疫组织化学法检测滑膜中外泌体标志物CD9的表达;分离培养原代RA-FLS和OA-FLS并提取上清液外泌体,使用Western印迹法和粒径分析(nanoparticle tracking analysis, NTA)鉴定;实时荧光定量PCR(RT-PCR)和CBA法分别检测OA-FLS和RA-FLS外泌体对M1和M2型极化分子mRNA水平及细胞因子分泌的影响。结果 RA关节液中M1型细胞因子IL-1β、IL-6、IL-8及M2型细胞因子IL-10水平均显著高于OA组;RA滑膜中CD9高表达;RA-FLS分泌的外泌体较OA-FLS明显增多;RA-FLS外泌体刺激M■后,细胞M1型极化分子mRNA表达和细胞因子分泌均增加。结论 RA-FLS外泌体促进M■向M1极化。Objective To investigate the role of exosomes derived from fibroblast-like synoviocytes from patients with rheumatoid arthritis(RA-FLS)in macrophage(M■)polarization.Methods Synovial fluid and tissue from osteoarthritis(OA)patients(n=6)and rheumatoid arthritis(RA)patients(n=6)were collected,and the cytokines in joint fluid were detected using flow cytometry bead array(CBA)method.The expression of extracellular marker CD9 in the synovium was detected with immunohi-stochemical method.Primary synovial RA-FLS and OA-FLS were isolated,and the exosomes from cell-culture supernatant were identified by Western blotting and nanoparticle tracking analysis(NTA).The effects of OA-FLS and RA-FLS exosomes on the M1 and M2 phenotype genes were determined with real time fluorescence quantitative PCR(qRT-PCR)and CBA.Results The expressions of M1 type markers(IL-1β,IL-6,IL-8),and M2 type marker(IL-10)in RA joint fluid were significantly higher than those in the OA group.High expression of CD9 was detected in RA synovium.RA-FLS was found to secrete more exosomes than OA-FLS.Elevated gene and protein expression levels of M1 markers were found in M■ treated with RA-FLS derived exosomes.Conclusion RA-FLS derived exosomes can induce M1 polarization in vitro.

关 键 词：类风湿关节炎 滑膜成纤维细胞 巨噬细胞 外泌体 M1型极化 

分 类 号：R392.12[医药卫生—免疫学]