MTX联合复方风湿宁对RA滑膜成纤维细胞IL-22的影响

Effect of MTX with compound Fengshining on synovial membrane fibroblasts IL-22 of patients with RA

目的观察甲氨蝶呤(MTX)和风湿宁对类风湿性关节炎滑膜成纤维细胞IL-22水平的影响,为其治疗机制的研究提供新靶点。方法分离、纯化、培养类风湿性关节炎患者滑膜组织成纤维样滑膜细胞,并分别加入MTX(MTX组)、复方风湿宁(复方风湿宁组)、MTX联合复方风湿宁(联合组)进行共培养,MTT法检测MTX和复方风湿宁对滑膜成纤维细胞增殖的影响;用RT-PCR方法检测各组滑膜成纤维细胞IL-22及凋亡相关基因caspase-3和caspase-9的mRNA表达。结果联合组的细胞增殖抑制率和caspase-3、caspase-9 mRNA表达水平明显高于其他两组(P<0.05),IL-22 mRNA表达水平明显低于其他两组(P<0.05),但MTX组和复方风湿宁组之间各项指标无显著差异(P>0.05)。结论 MTX和复方风湿宁均可降低类风湿性关节炎滑膜成纤维细胞IL-22水平,抑制细胞增殖反应,且联合应用可发挥协同效果,增强疗效。

Objective To observe the effect of methotrexate （ MTX ） and compound Fengshining （ FSN ） on synovial membrane fibroblasts IL-22 of patients with rheumatoid arthritis （ RA ） in order to provide new targets for the research on the therapeutic mechanism .Methods Synovial membrane fibroblasts of the patients with RA were obtained after separation , purification , and cultivation;and then, MTX （MTX group）, compound FSN （compound FSN group）, and MTX with compound FSN （combination group ） were added respectively for further cultivation .MTT method was used to determine the effect of MTX and compound FSN on the proliferation of synovial membrane fibroblasts , RT-PCR method was used to determine the expression level of IL-22 and that of apoptosis-related caspase-3 and caspase-9 mRNA in every group .Results The inhibitory rate of cell proliferation and expression levels of caspase-3 and caspase-9 mRNA in the combination group were significantly higher than those in the other two groups （P〈0.05）, and the expression level of IL-22 mRNA was significantly lower than that in the other two groups （P〈0.05）, but there was no significant difference in other indexes between the other two groups （ P〉0.05）.Conclusion Both MTX and compound FSN can reduce the level of synovial membrane fibroblasts IL-22 of patients with RA and inhibit the cell proliferation reaction , and combination application of them can obtain a synergistic effect and enhance the efficacy .