甲氨蝶呤对类风湿关节炎CCP/AST分泌白介素-4及干扰素-γ水平的影响

Effects of methotrexate on the secretion of interferon-γ and interleukin-4 by rheumatoid arthritis′ circum-citrullinated peptide antigen specific T cell

目的：观察甲氨蝶呤（methotrexate,MTX）对类风湿关节炎（rheumatoid arthritis,RA）患者外周血环瓜氨酸肽抗原特异性T细胞（circum-citrullinated peptide antigen specific Tcells,CCP/AST）分泌白介素-4（IL-4）及干扰素-γ（IFN-γ）水平的影响。方法：体外分离、培养RA患者外周血淋巴细胞（peripheral blood lymphocyte,PBL）,分别设RA空白组不加任何干预药物;RA CCP组仅加CCP干预（CCP终浓度为20μg/ml）;RAMTX组同时加CCP与MTX干预（CCP与MTX终浓度均为20μg/ml）;健康对照组培养健康者PBL,不加任何干预药物。体外培养72 h后,采用酶联免疫吸附法检测细胞培养上清中IL-4、IFN-γ分泌水平。结果：CCP干预后,RA患者PBL分泌的IFN-γ水平升高,IL-4分泌水平下降,IFN-γ/IL-4比值也明显升高;加入MTX后,RA患者CCP/AST分泌的IFN-γ水平下调,IL-4水平升高,IFN-γ/IL-4比值下降,差异有统计学意义（P〈0.05~P〈0.01）。结论：MTX对RA患者PBL源CCP/AST分泌的TH1/TH2型细胞因子的网络失衡状态有调节作用;MTX可以抑制CCP诱导的TH1型细胞因子,上调TH2型细胞因子的表达,这可能是MTX治疗RA有效途径之一。

Objective：To observe the effects of methotrexate（MTX） on the secretion of interferon-γ（IFN-γ） and interleukin-4（IL-4） by rheumatoid arthritis′ circum-citrullinated peptide antigen specific T cells（CCP/AST）.Methods：The rheumatoid arthritis′ blood peripheral lymphocytes（PBLs） were cultured in vitro for 72 h,and divided into three groups：lymphocytes only（RA blank group）,lymphocytes with CCP at the final concentration of 20 μg/ml（RA CCP group）,lymphocytes with CCP and MTX both at the final concentration of 20 μg/ml（RA MTX group）.PBLs from health volunteers were cultured in vitro（abbreviated blank control group）.The concentrations of IFN-γ and IL-4 were measured by ELISA.Results：Compared with RA CCP group：IFN-γ level of PBLs was lower and IL-4 was higher in RA MTX group,IFN-γ/IL-4 ratio was down-regulated（P0.05-P0.01）,which had the statistical significances.Conclusions：MTX can regulate the levels of IFN-γ and IL-4 in rheumatoid arthritis′ CCP/AST（P0.05）.MTX can inhibit CCP induced TH1 cytokine,and up-regulate TH2 cytokine expression.The regulation of rheumatoid arthritis′ CCP/AST by MTX may be one of the effective pathway in treating arthritis rheumatoid.