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.
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。
Keywords: inflammation; miR-342-3p; migration; rheumatoid arthritis.