类风湿性关节炎患者滑膜成纤维细胞来源的外泌体可促进巨噬细胞迁移，而这种迁移可被miR-124-3p抑制。
Exosomes derived from synovial fibroblasts from patients with rheumatoid arthritis promote macrophage migration that can be suppressed by miR-124-3p.
作者信息
Nakamachi Yuji, Uto Kenichi, Hayashi Shinya, Okano Takaichi, Morinobu Akio, Kuroda Ryosuke, Kawano Seiji, Saegusa Jun
机构信息
Department of Clinical Laboratory, Kobe University Hospital, 7-5-2, Kusunoki cho, Chuo Ku, Kobe 650-0017, Japan.
Department of Orthopaedic Surgery, Kobe University Graduate School of Medicine, 7-5-1, Kusunoki cho, Chuo Ku, Kobe 650-0017, Japan.
出版信息
Heliyon. 2023 Mar 30;9(4):e14986. doi: 10.1016/j.heliyon.2023.e14986. eCollection 2023 Apr.
OBJECTIVES
Exosomes are potent vehicles for intercellular communication. Rheumatoid arthritis (RA) is a chronic systemic disease of unknown etiology. Local administration of miR-124 precursor to rats with adjuvant-induced arthritis suppresses systemic arthritis and bone destruction. Thus, exosomes may be involved in this disease. We aimed to determine the role of exosomes in the pathology of RA.
METHODS
Fibroblast-like synoviocytes (FLS) were collected from patients with RA and osteoarthritis (OA). miR-124-3p mimic was transfected into the RA FLS (RA miR-124 FLS). Exosomes were collected from the culture medium by ultracentrifugation. Macrophages were produced from THP-1 cells. MicroRNAs in the exosomes were analyzed using real-time PCR. Proteomics analysis was performed using nanoscale liquid chromatography-tandem mass spectrometry. Macrophage migration was evaluated using a Transwell migration assay. SiRNA was used to knockdown proteins of interest.
RESULTS
MicroRNAs in the RA FLS, RA miR-124 FLS, and OA FLS exosomes were similar. Proteomics analysis revealed that pentraxin 3 (PTX3) levels were higher in RA FLS exosomes than in RA miR-124 FLS and OA FLS exosomes, and proteasome 20S subunit beta 5 (PSMB5) levels were lower in RA FLS exosomes than in RA miR-124 FLS and OA FLS exosomes. The RA FLS exosomes promoted and the RA miR-124 FLS exosomes suppressed macrophage migration. PTX3-silenced RA FLS exosomes suppressed and PSMB5-silenced OA FLS exosomes promoted macrophage migration.
CONCLUSIONS
RA FLS exosomes promote macrophage migration via PTX3 and PSMB5, and miR-124-3p suppresses this migration.
目的
外泌体是细胞间通讯的有效载体。类风湿关节炎（RA）是一种病因不明的慢性全身性疾病。向佐剂诱导性关节炎大鼠局部施用miR-124前体可抑制全身性关节炎和骨质破坏。因此，外泌体可能参与了这种疾病。我们旨在确定外泌体在RA病理中的作用。
方法
从类风湿关节炎患者和骨关节炎（OA）患者中收集成纤维样滑膜细胞（FLS）。将miR-124-3p模拟物转染到类风湿关节炎FLS（RA miR-124 FLS）中。通过超速离心从培养基中收集外泌体。巨噬细胞由THP-1细胞产生。使用实时PCR分析外泌体中的微小RNA。使用纳米级液相色谱-串联质谱进行蛋白质组学分析。使用Transwell迁移试验评估巨噬细胞迁移。使用小干扰RNA敲低感兴趣的蛋白质。
结果
类风湿关节炎FLS、RA miR-124 FLS和骨关节炎FLS外泌体中的微小RNA相似。蛋白质组学分析显示，类风湿关节炎FLS外泌体中的五聚体蛋白3（PTX3）水平高于RA miR-124 FLS和骨关节炎FLS外泌体，而类风湿关节炎FLS外泌体中的蛋白酶体20S亚基β5（PSMB5）水平低于RA miR-124 FLS和骨关节炎FLS外泌体。类风湿关节炎FLS外泌体促进巨噬细胞迁移，而RA miR-124 FLS外泌体抑制巨噬细胞迁移。PTX3沉默的类风湿关节炎FLS外泌体抑制巨噬细胞迁移，PSMB5沉默的骨关节炎FLS外泌体促进巨噬细胞迁移。
结论
类风湿关节炎FLS外泌体通过PTX3和PSMB5促进巨噬细胞迁移，而miR-124-3p抑制这种迁移。
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