类风湿关节炎患者CD4^(+)T淋巴细胞中miR-146a的表达及其与炎性细胞因子的相关性

Expression of miR-146a in CD4^(+)T lymphocytes of patients with rheumatoid arthritis and its correlation with inflammatory cytokines

目的·探讨类风湿关节炎(rheumatoid arthritis,RA)患者外周血CD4^(+)T淋巴细胞中miR-146a的表达,及其与炎性细胞因子[肿瘤坏死因子α(tumor necrosis factor-α,TNF-α)、白细胞介素-6(interleukin-6,IL-6)]的相关性。方法·选择2019年8月—2021年7月期间在深圳市龙华区人民医院就诊的活动期RA患者30例,并选择同期在该院体检的健康人群30例为健康对照者。分别抽取2组被试者的静脉血,先后分离获取外周血单个核细胞及CD4^(+)T淋巴细胞。采用实时荧光定量PCR(quantitative real-time PCR,qPCR)检测2组被试者外周血CD4^(+)T淋巴细胞中miR-146a的表达,并采用酶联免疫吸附试验(enzyme-linked immunosorbent assay,ELISA)对其TNF-α、IL-6水平进行检测。使用miR-146a mimic转染RA患者的外周血CD4^(+)T淋巴细胞,并于转染后再次对miR-146a、TNF-α及IL-6的表达进行检测。采用Pearson相关系数对转染前、后RA患者的miR-146a表达与TNF-α、IL-6表达的相关性进行分析。结果·转染miR-146a mimic前,与健康对照者相比,RA患者外周血CD4^(+)T淋巴细胞中miR-146a、TNF-α及IL-6的表达较高(均P<0.001)。转染后RA患者外周血CD4^(+)T淋巴细胞中miR-146a的表达水平较转染前有所升高、TNF-α及IL-6的表达则有所下降(均P<0.001)。Pearson相关分析的结果显示,转染前RA患者外周血CD4^(+)T淋巴细胞中miR-146a的表达水平与TNF-α、IL-6的表达水平呈正相关(r=0.959,P<0.001;r=0.916,P<0.001),转染后其miR-146a的表达水平与TNF-α、IL-6的表达水平亦呈正相关(r=0.971,P<0.001;r=0.861,P<0.001)。结论·miR-146a可能通过影响RA患者外周血CD4^(+)T淋巴细胞分泌的TNF-α、IL-6等相关炎性细胞因子,在一定程度上参与RA的发病过程。

Objective·To investigate the expression of miR-146a in peripheral blood CD4^(+)T lymphocytes of patients with rheumatoid arthritis(RA)and its correlation with inflammatory cytokines such as tumor necrosis factor-α(TNF-α)and interleukin-6(IL-6).Methods·A total of 30 active RA patients who received medical treatment and 30 healthy controls who underwent physical examinations at the People's Hospital of Longhua,Shenzhen from August 2019 to July 2021 were selected.Peripheral blood mononuclear cells(PBMCs)and CD4^(+)T lymphocytes were isolated from venous blood extracted from RA patients and healthy controls,respectively.Quantitative real-time PCR(qPCR)was used to detect the expression of miR-146a in peripheral blood CD4^(+)T lymphocytes,and enzyme-linked immunosorbent assay(ELISA)was used to detect the levels of TNF-αand IL-6.After transfection of the peripheral blood CD4^(+)T lymphocytes of RA patients with miR-146a mimic,the expression of miR-146a,TNF-αand IL-6 was detected again.The correlations between miR-146a expression and TNF-αand IL-6 expression in RA patients,both before and after transfection,were analyzed by using Pearson correlation coefficient.Results·Before transfection with miR-146a mimic,the expression levels of miR-146a,TNF-αand IL-6 in peripheral blood CD4^(+)T lymphocytes of RA patients were significantly higher than those of healthy controls(all P<0.001).After transfection,the expression of miR-146a in peripheral blood CD4^(+)T lymphocytes of RA patients was significantly higher,and the expression of TNF-αand IL-6 was significantly lower(all P<0.001).The results of Pearson correlation analysis showed that the expression of miR-146a in peripheral blood CD4^(+)T lymphocytes of RA patients,both before and after transfection,was positively correlated with the expression of TNF-αand IL-6,respectively(r=0.959,P<0.001;r=0.916,P<0.001;r=0.971,P<0.001;r=0.861,P<0.001).Conclusion·miR-146a can regulate the levels of TNF-αand IL-6 in peripheral blood CD4^(+)T lymphocytes of RA patients,indicating that miR-146a may play a role in the pathogenesis of RA.