目的 研究滑液（SF）外泌体 miRNA 在骨关节炎（OA）患者中的作用并探讨其潜在分子机制。收集男性和女性 OA 患者的退变膝关节组织。采用酶联免疫吸附试验（ELISA）检测退变组和损伤组炎症指标TNF-α、IL-6、IL-10表达的差异。使用 Exoquick 试剂盒从 SF 中分离外泌体，并使用微阵列来识别差异表达的 miRNA (DEmiRNA)，并使用生物信息学对其进行分析。使用荧光素酶报告基因测定验证了 DEmiRNA 和靶基因之间的预测关系。 CCK-8 和 Transwell 测定用于评估细胞活力和迁移。采用免疫荧光和TUNEL法检测细胞自噬和凋亡。通过免疫沉淀检测蛋白质之间的相互作用，并通过Mab救援实验验证蛋白质之间的相互作用。变性组中TNF-α/IL6的相对表达量显着高于损伤组。 OA变性组释放的外泌体明显多于损伤组且更小。 miR-182-5p在OA患者中表达显着降低，且与炎症指标相关性较高。肿瘤坏死因子α诱导蛋白8（TNFAIP8）是miR-182-5p的靶标，其过度表达促进软骨细胞增殖、迁移和侵袭，提高伤口愈合效率。我们还发现 TNFAIP8 与自噬相关基因 3 (ATG3) 存在直接相互作用。 TNFAIP8 触发 ATG3 LC3 介导的自噬。外泌体 miR-182-5p 的下调通过 ATG/LC3 途径靶向 TNFAIP8 抑制 OA 变性。版权所有 © 2023。由 Elsevier B.V. 出版。

To investigate the role of exosomal miRNAs from synovial fluid (SF) in osteoarthritis (OA) patients and investigate the underlying molecular mechanism.Degenerated knee tissues were collected from male and female OA patients. Enzyme-linked immunosorbent assay (ELISA) was used to detect the differences in the expression of inflammatory indicators, including TNF-α, IL-6, and IL-10, between the degenerative and injury groups. Exosomes were isolated from SF using the Exoquick kit, and a microarray was used to identify differentially expressed miRNAs (DEmiRNAs), which were analyzed using bioinformatics. The predicted relationship between DEmiRNAs and target genes was verified using a luciferase reporter gene assay. CCK-8 and transwell assays were used to assess cell viability and migration. Immunofluorescence and TUNEL assay were used to detect cell autophagy and apoptosis. The interaction between proteins was detected by immunoprecipitation and verified by Mab rescue assay.The relative expression of TNF-α/IL6 was significantly higher in the degeneration group than in the injury group. The OA degeneration group released significantly more and smaller exosomes than the injury group. The expression of miR-182-5p was markedly reduced in OA patients and had a higher correlation with inflammatory indicators. Tumor necrosis factor α-induced protein 8 (TNFAIP8) was a target of miR-182-5p, and its overexpression promoted chondrocyte proliferation, migration, and invasion and enhanced the wound healing efficiency. We also found a direct interaction of TNFAIP8 with autophagy-related gene 3 (ATG3). TNFAIP8 triggered ATG3 LC3-mediated autophagy.The downregulation of exosomal miR-182-5p inhibits OA degeneration by targeting TNFAIP8 via the ATG/LC3 pathway.Copyright © 2023. Published by Elsevier B.V.