1,25-二羟维生素D3对初诊类风湿关节炎患者外周血单个核细胞分泌Th17相关细胞因子的影响

1, 25 （OH）2-Vitamin-D3 attenuates Th17-related cytokines expression in peripheral blood mononuclear cells in patients with early-diagnosed rheumatoid arthritis

目的 探讨1,25（OH）2D3对初诊类风湿关节炎（RA）患者辅助性T淋巴细胞（Th）17相关细胞因子的作用及其治疗RA潜在机制.方法 选初诊RA患者18例（RA组）,健康对照18例（对照组）.分离2组受试者外周血单个核细胞（PBMC）,加抗CD3和CD2s刺激,1,25（OH）2D3和/或甲氨蝶呤共培养,流式细胞仪微球捕获芯片技术（CBA）检测IL-17、IL-6、TNFα水平,ELISA检测IL-22水平.结果 RA组患者血清IL-17[（43 ±6） ng/L]、TNFα[（5.91 ±2.53）ng/L]、IL-6[（16.6±12.0）ng/L]、IL-22[（51±17） ng/L]水平显著高于对照组[（21±3） ng/L,（2.63±0.27） ng/L,（4.2±2.3） ng/L,（22±14） ng/L],差异有统计学意义（P均＜0.05）.RA组PBMC经不同浓度1,25（OH）2D3处理IL-17、TNFα、IL-6水平与未处理比差异有统计学意义（P均＜0.05）,不同浓度间比较差异无统计学意义（P均＞0.05）;IL-22水平均有下降趋势,但差异无统计学意义（P均＞0.05）.RA组PBMC经不同浓度甲氨蝶呤处理IL-17、TNFα水平与未处理比差异有统计学意义（P均＜0.05）,不同浓度间比较差异无统计学意义（P均＞0.05）;IL-22、IL-6差异无统计学意义（P均＞0.05）.RA组PBMC经1 nmoL/L 1,25（OH）2D3和0.5 mg/ml甲氨蝶呤处理后,与未处理比,IL-17、IL-22、IL-16、TNFα差异有统计学意义（P均＜0.05）.结论 1,25（OH）2D3作为一种免疫调节剂,对Th17相关细胞因子具有抑制作用.

Objective To investigate the effects of 1,25-dihydroxyvitamin D3 [1,25 （OH） 2D3] on T helper cell type 17 （Th17） cytokines and therapeutic mechanism in patients with rheumatoid arthritis （RA）.Methods Peripheral blood mononuclear cells （PBMCs） from healthy donors and RA patients were collected.The PBMCs were stimulated with anti-CD3/anti-CD28 monoclonal antibodies in the absence or presence of 1,25（OH）2D3 and methotrexate （MTX）.After co-culture,the serum levels of Th17 cytokines interleukin （IL）-17,IL-6,tumour necrosis factor alpha （TNFα） were analyzed by cytometric bead array （CBA）.The level of IL-22 was analyzed by enzyme-linked immunosorbent assay （ELISA）.The independent samples t test and one-way analysis of variance （ANOVA） were used for statistical analysis.Results The levels of cytokines IL-17,TNFα,IL-6 and IL-22 in RA group were significantly higher than those in the control group[（43 ± 6） ng/L,（5.91 ± 2.53） ng/L,（16.6 ± 12.0） ng/L,（51 ± 17） ng/L vs （21 ±3）ng/L,（2.63 ±0.27） ng/L,（4.2 ±2.3） ng/L,（22 ± 14） ng/L].Each of the three different 1,25 （OH） 2 D3 doses inhibited secretion of IL-17 [（533 ± 47） pg/ml,（426 ± 55） pg/ml,（319 ± 86） pg/ml],TNFα[（424 ± 82） pg/ml,（382 ± 79） pg/ml,（326 ± 87） pg/ml],and IL-6 [（5 513 ± 3 429） pg/ml,（4 555 ±3 157）pg/ml,（3 748 ± 1 919）pg/ml]in RA group （P 〈0.05）,yet no statistical difference was found in IL-22 secretion with a trend of decrease after treatment of 1,25 （OH）2D3.Three different doses of MTX inhibited secretion of IL-17 [（452 ± 50） pg/ml,（372 ± 67） pg/ml,（315 ± 104） pg/ml] and TNFα [（319 ± 74） pg/ml,（292 ± 59） pg/ml,（266 ± 64） pg/ml] in RA group （P 〈 0.05）.However,levels of IL-6 and IL-22 were not affected after treated with MTX.Conclusion Our data indicated that 1,25 （OH）2D3 may play as an immune modulating agent to suppress Th17 cell cytokines.Supplement of vitamin D has the effective potential to treat patients with RA or other Th17 cell mediated autoimmune disorders.