类风湿关节炎患者TH17细胞及其相关细胞因子的表达及临床意义

Clinical significance of TH17 cells and related cytokines expression in RA patients

关键词:  类风湿性关节炎  TH17细胞  IL-17  IL-17mRNA  RT-PCR  流式细胞仪

Key words:  Rheumatoid arthritis (RA)  TH17 cells  IL-17  IL-17mRNA  RT-PCR  flow cytometry

摘要:

目的　探讨类风湿关节炎（RA）患者TH17细胞及其相关细胞因子的表达及临床意义。方法　收集大连医科大学附属第一医院门诊RA初治活动期患者的外周血标本32例为活动期RA组，健康献血者30例为健康对照组，所有受试者清晨空腹坐位肘静脉采血。应用酶联免疫吸附法(ELISA)及反转录PCR(RT-PCR)法，分别检测外周血血清中IL-17水平和外周血单个核细胞(PBMC)中IL-17mRNA表达水平；将PBMC一组不加刺激，另一组用佛波酯(50 ng/mL)和离子霉素(1 μg/mL)刺激4 h后，流式细胞仪检测TH17细胞数量。结果　活动期RA患者血清IL-17水平(15.76±0.62) pg/mL与健康对照组(8.67±0.12) pg/mL比较，差异有显著性意义(P<0.05)。活动期RA患者PBMC的IL-17mRNA表达水平(1.94±0.46)与健康对照组(0.84±0.16)比较，差异有显著性意义(P<0.05)。RA组血清IL-17水平与mRNA表达水平呈正相关(r=0.70, P<0.05)，且与疾病活动程度(DAS28)评分呈正相关(r=0.99, P<0.05)。活动期RA患者外周血TH17细胞比例(2.50±0.54)%与健康对照组(0.03±0.00)%比较，差异有显著性意义(P<0.05)。但刺激组(3.23±0.05)%与无刺激组(2.50±0.54)%比较，差异无显著性意义(P>0.05)。RA患者组TH17细胞数量与疾病活动程度(DAS28)评分呈正相关(r=0.96,P<0.05)。结论　活动期RA患者TH17细胞及其主要效应性细胞因子IL-17在细胞、蛋白和基因表达水平上均明显升高并与疾病活动程度呈正相关，对临床制定治疗方案、判断疗效有指导意义。

Abstract:

Objective　To investigate TH17 cells and their cytokine expression in rheumatoid arthritis patients, which may play a positive role in clarifying the pathogenesis of RA. Methods　Utilizing Enzyme-linked immunosorbent assay and reverse transcription PCR methods, IL-17 protein in serum and mRNA in peripheral blood mononuclear cells (PBMCs) were measured in RA patients. TH17 cells were detected by flow cytometry in a group without stimulation and the other group with phorbol ester (50 ng/mL) and ionomycin (1 μg/mL) stimulation for 4 hours. Results　Serum IL-17 levels in active RA patients were significantly higher than those in the normal control group (15.76±0.62) pg/mL vs. (8.67±0.12) pg/mL (P<0.05). PBMC IL-17 mRNA expression in active RA patients were significantly higher than the normal control group (1.94±0.46) vs. (0.84±0.16) (P<0.05). IL-17 protein levels were positively associated with mRNA expression levels (r=0.70, P<0.05). Serum IL-17 level of RA patients and disease activity (DAS28) score were positively correlated (r=0.99, P<0.05). In the peripheral blood of RA patients, TH17 cells proportion increased (2.50±0.54)% vs. (0.03±0.00)% (P<0.01), and there was no difference between stimulation and non-stimulation groups (3.23±0.05)% vs. (2.50±0.54)% (P>0.05). TH17 cells of RA patients and disease activity (DAS28) score were positively correlated (r=0.96, P<0.05). Conclusion　In active RA patients, TH17 cells and their cytokine expression are significantly increased in the levels of cell, protein and mRNA, which suggests that TH17 cells might play an important role in the pathogenesis of RA.