miR-146a调控TLRs信号通路对类风湿关节炎患者外周血单个核细胞的影响

The Effect of miR-146a Regulating TLRs Signaling Pathway on Peripheral Blood Mononuclear Cells in Patients with Rheumatoid Arthritis

目的:观察miR-146a调控TLRs信号通路对活动期类风湿关节炎患者外周血单个核细胞的影响。方法:选取20例活动期类风湿关节炎患者,对患者外周血单个核细胞分离培养,分为LPS组和LPS+miR-146a组;同时选取10例健康体检者为健康对照组。采用MTT法测定吸光度,比较外周血单个核细胞变化,检测外周血单个核细胞的凋亡情况。RT-PCR法检测外周血单个核细胞miR-146a、Toll样受体2(TLR2)、髓样分化因子88(MyD88)mRNA表达。Western blot法检测外周血单个核细胞miR-146a、TLR2、MyD88蛋白表达。结果:LPS组较健康对照组和LPS+miR-146a组外周血单个核细胞TLR2表达明显升高(P<0.05);LPS组较健康对照组和LPS+miR-146a组外周血单个核细胞中miR-146a、TLR2、MyD88 mRNA表达明显升高(P<0.05);LPS组较健康对照组和LPS+miR-146a组外周血单个核细胞miR-146a、TLR2、MyD88蛋白表达明显升高(P<0.05)。结论:高表达miR-146a可能通过依赖TLR2/MyD88信号通路,抑制活动期类风湿关节炎患者外周血单个核细胞中TLR2、MyD88表达,减轻炎症反应。

Objective:To observe the effect of miR-46 regulation of TLRs signaling pathway on peripheral blood mononuclear cells in patients with active rheumatoid arthritis.Methods:Twenty patients with active rheumatoid arthritis were selected and divided into the LPS group and the LPS+miR-146a group,and peripheral blood mononuclear cells in them were isolated and cultured.At the same time,ten healthy examinees were selected as the healthy control group.MTT method was used to measure absorbance,changes in peripheral blood mononuclear cells were compared,and apoptosis of peripheral blood mononuclear cells was detected.RT-PCR was used to detect the mRNA expression of miR-146a,TLR2,and MyD88 in peripheral blood mononuclear cells.Protein immunoblotting was used to detect the protein expression of miR-146a,TLR2,and MyD88 in peripheral blood mononuclear cells.Results:The TLR2 expression rate of peripheral blood mononuclear cells in the LPS group was significantly increased compared to the healthy control group and the LPS+miR-146a group(P<0.05);the expression of protein mRNA of miR-146a,TLR2,and MyD88 in peripheral blood mononuclear cells of the LPS group was significantly increased compared to the healthy control group and the LPS+miR-146a group(P<0.05);the protein levels of miR-146a,TLR2,and MyD88 in peripheral blood mononuclear cells were significantly increased in the LPS group compared to the healthy control group and the LPS+miR-146a group(P<0.05).Conclusion:High expression may inhibit the expression of TLR2 and MyD88 in peripheral blood mononuclear cells of patients with active rheumatoid arthritis by relying on the TLR2/MyD88 signaling pathway,reducing inflammatory response.