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

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

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。

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 miR342-3p,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-αusing 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-αstimulation 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.