来氟米特活性代谢物teriflunomide联合甲氨蝶呤对类风湿关节炎成纤维样滑膜细胞的影响  

Effect of Teriflunomide Combined with Methotrexate on Fibroblast-like Synoviocytes in Rheumatoid Arthritis

作　　者：姚瑶[1] 葛卫红[1] 

机构地区：[1]南京大学医学院附属鼓楼医院药学部,南京210008

出　　处：《医药导报》2013年第7期839-842,共4页Herald of Medicine

基　　金：南京市卫生局医学科技发展项目(ZKX08022;YKK11101);江苏省六大人才高峰计划项目(2010-057);南京市鼓楼医院青年启动项目(QJ2012020)

摘　　要：目的观察来氟米特活性代谢物teriflunomide联合甲氨蝶呤对体外培养类风湿关节炎患者成纤维样滑膜细胞增殖及其破骨细胞分化因子表达的影响。方法滑膜组织分离培养滑膜成纤维细胞,加入不同浓度药液,使得终浓度分别为0.1,0.01,0.001 mg.mL-1 teriflunomide,0.1,0.01,0.001 mg.mL-1甲氨蝶呤,0.1 mg.mL-1 teriflunomide+0.1 mg.mL-1甲氨蝶呤,0.01 mg.mL-1 teriflunomide+0.01 mg.mL-1甲氨蝶呤,0.001 mg.mL-1 teriflunomide+0.001 mg.mL-1甲氨蝶呤,每组重复3个复孔,四甲基偶氮唑盐(MTT)法检测药物对细胞增殖的影响;半定量逆转录-聚合酶链反应(RT-PCR)检测药物对滑膜成纤维细胞核因子-κB(NF-κB)受体活化因子配体、骨保护素mRNA表达水平的影响。结果与对照组比较,药物处理后滑膜成纤维细胞的吸光度(A)值均有所降低,与单独用药组比较,0.1 mg.mL-1teriflunomide联合0.1 mg.mL-1甲氨蝶呤组A值降低最为显著(P<0.05),且该处理组骨保护素mRNA表达升高,NF-κB受体活化因子配体mRNA表达降低,与对照组比较差异有统计学意义(P<0.05)。结论来氟米特活性代谢物teriflunomide与甲氨蝶呤联合能够协同抑制滑膜成纤维细胞增殖,调节骨破坏相关基因表达,这可能是来氟米特与甲氨蝶呤联用治疗类风湿性关节炎的机制之一。Objective To investigate the effects of leflunomide＇s active metabolite teriflunomide （TEF） in combination with methotrexate （MTX） on the proliferation of fibroblast-like synovioeytes （FLS）, as well as the expression of osteoelast differentiation factor in FLS. Methods FLS were isolated from the synoviuin of patients with rheumatoid arthritis, and were incubated with different concentrations of TEF, MTX, or both combined. MTT assay was applied to detect the growth of FLS. The expression of RANKL and OPG mRNA in FLS was observed by seml-quantified RT-PCR assay. Results The medications significantly decreased the absorbance value in the FLS culture as compared to the control group （P〈0.05）. The absorbance in the culture treated with the combination of TEF （0.1 mg ~ mL-1 ） and MTX （0.1 mg ~ mL-1 ） was significantly lower than the mono-treated group （P〈0.05）. Moreover, OPG mRNA expression was significantly enhanced while the expression of RANKL mRNA decreased in these groups, compared with the control group （P 〈 0. 05 ）. Conclusion TEF and MTX have a synergistic effect in inhibiting FLS. The combination of TEF and MTX inhibits FLS proliferation and modulates the expression of osteoclast differentiation factors such as RANKL and OPG, which exolains the inhibitory mechanisms of bone damage in RA.

关 键 词：来氟米特活性代谢物 甲氨蝶呤 类风湿关节炎 成纤维样滑膜细胞 

分 类 号：R971.1[医药卫生—药品] R965[医药卫生—药学]