南方医科大学学报

南方医科大学学报 ›› 2022, Vol. 42 ›› Issue (11): 1712-1719.doi: 10.12122/j.issn.1673-4254.2022.11.16

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MiR-342-3p在类风湿关节炎患者中低表达并促进滑膜成纤维细胞的炎症和迁移

周 琴，刘 健，孙艳秋，陈晓露，张先恒，丁 香   

  1. 安徽中医药大学第一附属医院，安徽 合肥 230012
  • 出版日期:2022-11-20 发布日期:2022-11-30

Expression of miR-342-3p in rheumatoid arthritis patients and its effect on synovial fibroblast inflammation and migration

ZHOU Qin, LIU Jian, SUN Yanqiu, CHEN Xiaolu, ZHANG Xianheng, DING Xiang   

  1. First Affiliated Hospital of Anhui University of Traditional Chinese Medicine, Hefei 230012, China
  • Online:2022-11-20 Published:2022-11-30

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摘要： 目的 探究类风湿关节炎(RA)患者miR-342-3p的表达，及其对类风湿性关节炎滑膜成纤维细胞细胞（RA-FLS）炎症和迁移的影响。方法 收集正常人30例、RA患者50例外周血单个核细胞（PBMCs），检测PBMCs中miR-342-3p的表达，并研究其与临床指标RF、ESR、anti-CCP、hs-CRP、C3、DAS-28、SAS、SDS的相关性。建立RA-FLS细胞系，20 ng/mL TNF-α刺激细胞，构建mimics-miR-342-3p 和 inhibitor-miR-342-3p 及空转组，转染至 RA-FLS 中；实验分为 6 组：RA-FLS 组，TNF-α+RA-FLS 组，mimics-NC组，mimics-miR-342-3p组，inhibitor-NC组和inhibitor-miR-342-3p组；采用定量实时聚合酶链反应（RT-qPCR）检测miR-342-3p的表达；酶联免疫吸附法（ELISA）检测白介素-1β（IL-1β）、白介素-6（IL-6）、白介素-10（IL-10）、肿瘤坏死因子-α（TNF-α）的表达。CCK8检测细胞活力。Transwell小室检测滑膜细胞的迁移。结果 与正常组相比，RA患者miR-342-3p表达显著降低（P<0.05），ROC曲线结果显示AUC 97.53% 。miR-342-3p与RF（r=-0.321）、ESR（r=-0.284）、anti-CCP（r=-0.355）、hs-CRP（r=-0.320）、C3（r=-0.294）、DAS-28（r=-0.395）、SAS（r=-0.366）、SDS（r=-0.397）呈显著负相关（P<0.05）。miR-342-3p与anti-CCP 、DAS-28、SDS、SAS的升高有较强的关联，规则支持均大于85%、置信度均大于88%和提升均大于1；与RA-FLS组相比，TNF-α刺激后细胞活力显著升高（P<0.05），IL-1β、IL-6、TNF-α表达显著升高，IL-10的表达显著降低（P<0.05）；与mimics-NC组相比，mimics-miR-342-3p组细胞活力显著降低（P<0.05）；与inhibitor-NC组相比，inhibitor-miR-342-3p组细胞活力显著升高（P<0.05）；与mimics-NC组相比，mimics-miR-342-3p组IL-1β、IL-6、TNF-α表达显著降低，IL-10的表达显著升高（P<0.05）；与inhibitor-NC组相比，inhibitor-miR-342-3p组IL-1β、IL-6、TNF-α表达显著升高，IL-10的表达显著降低（P<0.05）。结论 miR-342-3p在RA患者中表达降低，通过促进RA-FLS炎症和迁移，参与RA的发病机制。

关键词: 类风湿关节炎；miR-342-3p；炎症；迁移

Abstract: Objective To investigate the expression level of miR-342-3p in peripheral blood mononuclear cells (PBMCs) of patients with rheumatoid arthritis (RA) and its effect on inflammatory response and migration of synovial fibroblasts in RA (RA-FLS). Methods PBMCs were collected from 30 healthy individuals and 50 RA patients for detecting the expression of miR-342-3p, and its correlation with the clinical indicators RF, ESR, anti-CCP, hs-CRP, C3, DAS-28, SAS, and SDS was analyzed. In RA-FLS cultures, the effect of transfection of miR-342-3p mimics and inhibitor on TNF-α-induced inflammatory response of the cells was evaluated by detecting the expressions of IL-1β, IL-6, IL-10, and TNF-α using ELISA. CCK8 assay and Transwell assay were used for detecting the changes in cell viability and migration ability of the synovial cells. Results In RA patients, the expression level of miR-342-3p was significantly lowered in the PBMCs (P<0.05) with an area under the ROC curve of 97.53% and showed inverse correlations with RF (r=-0.321), ESR(r=-0.284), anti-CCP (r=-0.355), hs-CRP (r=-0.320), C3 (r=-0.294), DAS-28 (r=-0.395), SAS (r=-0.366), and SDS (r=-0.397) (all P<0.05); a low expression of miR-342-3p was strongly associated with elevated levels of anti-CCP, DAS-28, SDS, and SAS (all with a rule support greater than 85%, confidence greater than 88%, and lift greater than 1). In cultured RA-FLS, TNF-α stimulation significantly increased the cell viability (P<0.05), upregulated the expressions of IL-1β, IL-6, and TNF-α, and lowered the expression of IL-10 (P<0.05). These changes were significantly suppressed by transfection of the cells with miR-342-3p mimics (P<0.05) but enhanced by transfection with miR-342-3p inhibitor (P<0.05). Conclusion The expression of miR-342-3p is decreased in the PBMCs of RA patients. A lowered expression of miR-342-3p contributes to the pathogenesis of RA by promoting inflammatory responses and migration of RA-FLS.

Key words: rheumatoid arthritis; miR-342-3p; inflammation; migration