外周血单个核细胞miR-146a和微管相关蛋白1轻链3及红细胞沉降率与类风湿关节炎疾病活动的关系

Correlations of miR-146a, LC3 and erythrocyte sedimentation rate with rheumatoid arthritis activity

目的观察类风湿关节炎(rheumatoid arthritis, RA)患者外周血单个核细胞miR-146a、微管相关蛋白1轻链3(microtubule-associated protein 1 light chain 3, LC3)表达及红细胞沉降率(erythrocyte sedimentation rate, ESR)变化,探讨其与RA疾病活动的关系。方法 RA患者105例为RA组,采用28处关节疾病活动度(disease activity score 28, DAS28)评分评估RA疾病活动度,分为疾病缓解组26例、低活动度组44例、中高活动度组35例。同期体检健康者105例为对照组。RA组于入院次日、对照组于体检日采集空腹静脉血,采用实时荧光定量PCR法检测外周血单个核细胞miR-146a、LC3 mRNA相对表达量,采用魏氏法检测ESR。比较RA组与对照组、不同疾病活动度RA患者外周血单个核细胞miR-146a、LC3 mRNA相对表达量及ESR;Pearson相关法分析外周血单个核细胞miR-146a、LC3 mRNA相对表达量及ESR与DAS28评分的相关性;绘制ROC曲线,评估外周血单个核细胞miR-146a、LC3 mRNA及ESR诊断RA疾病活动的价值。结果 RA组外周血单个核细胞miR-146a(1.24±0.36)、LC3 mRNA(1.72±0.31)相对表达量及ESR[(47.64±10.18)mm/h]均高于对照组[0.80±0.21、1.06±0.28、(12.24±3.86)mm/h](P<0.05)。疾病缓解组、低活动度组、中高活动度组外周血单个核细胞miR-146a(0.98±0.17、1.19±0.23、1.50±0.34)、LC3 mRNA(1.26±0.25、1.61±0.33、2.20±0.42)相对表达量及ESR[(24.26±7.02)、(42.97±12.25)、(70.88±16.39)mm/h]、DAS28评分[(1.24±0.31)、(2.86±0.20)、(6.59±2.11)分]均依次升高(P<0.05)。RA患者外周血单个核细胞miR-146a相对表达量(r=0.625,P<0.001)、LC3 mRNA相对表达量(r=0.740,P<0.001)及ESR(r=0.794,P<0.001)与DAS28评分均呈正相关。外周血单个核细胞miR-146a、LC3 mRNA、ESR分别以1.23、1.47、36.60 mm/h为最佳截断值,单独及联合诊断RA疾病活动的AUC分别为0.797(95%CI:0.708~0.869,P<0.001)、0.809(95%CI:0.721~0.879,P<0.001)、0.837(95%CI:0.752~0.902,P<0.001)、0.931(95%CI:0.865~0.971,P<0.001),灵敏度分别为64.56%、81.01%、78.48%、88.61%,特异度分别为84.62%、73.08%、76.92%、92.31%。结论外周血单个核细胞miR-146a、LC3表达及ESR升高与RA疾病活动度高有关,三者联合诊断RA患者疾病活动有较高价值。

Objective To observe the expressions of miR-146a,microtubule-associated protein 1 light chain 3(LC3)and erythrocyte sedimentation rate(ESR)in patients with rheumatoid arthritis(RA),and to investigate their correlations with RA activity.Methods A total of 105 RA patients(RA group)were evaluated the RA activity according to disease activity score 28(DAS28),and were divided into remission group(n=26),low-activity group(n=44),and medium-/high-activity group(n=35).Another 105 healthy subjects were selected as controls(control group).Fasting venous blood was collected in RA group on the second day after admission and control group on physical examination day.The relative expressions of miR-146 aand LC3 mRNA in peripheral blood mononuclear cells were detected by real-time fluorescence quantitative PCR,and ESR was detected by Westergren method.The relative expressions of miR-146 aand LC3 mRNA as well as the ESR were compared between RA group and control group,and among RA patients with different disease activity levels.Pearson correlation method was used to analyze the correlations of the relative expressions of miR-146 aand LC3 mRNA as well as ESR with DAS28 score.ROC curve was drawn to evaluate the values of miR-146 a,LC3 mRNA and ESR to the diagnosis of RA.Results The relative expressions of miR-146 aand LC3 mRNA as well as ESR were higher in RA group[1.24±0.36,1.72±0.31,47.64±10.18mm/h]than those in control group[0.80±0.21,1.06±0.28,(12.24±3.86)mm/h](P<0.05).The relative expressions of miR-146 aand LC3 mRNA in peripheral blood mononuclear cells,ESR level and DAS28 score increased gradually in turn in remission group[0.98±0.17,1.26±0.25,(24.26±7.02)mm/h,1.24±0.31],low-activity group[1.19±0.23,1.61±0.33,(42.97±12.25)mm/h,2.86±0.20]and medium-/high-activity group[1.50±0.34,2.20±0.42,(70.88±16.39)mm/h,6.59±2.11](P<0.05).The relative expressions of miR-146 a(r=0.625,P<0.001)and LC3 mRNA(r=0.740,P<0.001)in peripheral blood mononuclear cells as well as ESR(r=0.794,P<0.001)were positively correlated with DAS28 score in RA patients.When the optimal cut-offvalues of miR-146a,LC3 mRNA and ESR were1.23,1.47 and 36.60 mm/h,the AUCs of the single and combined detection of them three were 0.797(95%CI:0.708-0.869,P<0.001),0.809(95%CI:0.721-0.879,P<0.001),0.837(95%CI:0.752-0.902,P<0.001),and 0.931(95%CI:0.865-0.971,P<0.001),the sensitivities were 64.56%,81.01%,78.48%and 88.61%,and the specificities were 84.62%,73.08%,76.92%and 92.31%,respectively.Conclusion The increased expressions of miR-146 aand LC3 in peripheral blood mononuclear cells and the increased level of ESR are correlated with high RA disease activity,and the combined detection of them three has a high value to the diagnosis of disease activity in RA patients.