来氟米特对类风湿关节炎患者Th17细胞免疫调节的影响

Immunoregulation effects of leflunomide on Th17 cells in peripheral blood of rheumatoid arthritis in vitro

目的:探讨来氟米特活性代谢产物A771726对类风湿关节炎(RA)患者外周血Th17细胞的影响及意义。方法:取RA患者外周血单个核细胞(PBMC),采用免疫磁珠法阴选出CD4+T辅助细胞(helper T cell,Th),将CD4+T细胞分别与不同剂量的A771726(15、30、60μg/ml)和地塞米松(10-6、10-7、10-8mol/L)共培养48 h,流式细胞术(FCM)检测RA患者外周血Th17/Th百分比;应用实时荧光定量逆转录聚合酶链技术(即RT-PCR)检测IL-17、ROR-γt mRNA水平。结果:在体外,来氟米特对RA患者外周血CD4+T细胞胞内IL-17、ROR-γt mRNA表达无明显抑制作用,地塞米松可以使CD4+T细胞胞内IL-17、ROR-γt mRNA表达明显减少,随着药物浓度的增加抑制作用更加明显。来氟米特对Th17/Th无明显影响,地塞米松能降低Th17/Th细胞百分比。结论:来氟米特对RA患者外周血Th17细胞无明显影响作用,而地塞米松则可显著降低Th17细胞在Th细胞中的比例。

Objective： To investigate the effect and significance of A771726（activated metabolite of leflunomide） and dexamethasone on the expression of Th17 cells in patients with rheumatoid arthritis（RA）.Methods： Isolatating CD4＋T cells from PBMC by magnetic cell separator and then cultured with A771726（concentration respectively as 15,30,60 μg/ml） or dexamethasone（concentration respectively as 10-6,10-7,10-8mol/L） for 48 hours.Determination of the percentage of Th17/Th by flow cytometry.The expression level of IL-17 and ROR-γt mRNA was determined by RT-PCR.Results： There was no obvious effect of leflunomide on intracellular IL-17 or ROR-γt mRNA of CD4＋ T cells.The expression level of intracellular IL-17 and ROR-γt mRNA on CD4＋ T cells were reduced by dexamethasone and these effect were more obvious with the increase of drug concentration.Leflunomide had no obvious effect on the percentage of Th17 cells,but dexamethasone could decrease the percentage of Th17 cells. Conclusion： Leflunomide had no obvious effect on Th17 cells,but dexamethasone could evidently decrease the percentage of Th17 cells.