甲氨蝶呤联合复方风湿宁对类风湿性关节炎滑膜成纤维细胞Treg/Th17免疫平衡的影响  

Effect of compound Fengshining combined with methotrexate on Treg/Th17 immune balance in fibroblast-like synoviocytes of rheumatoid arthritis

作　　者：李晓佳[1] 赵迎杰[2] 孙广臣[3] 

机构地区：[1]桂林医学院免疫学教研室,广西桂林541000 [2]山西医科大学第二临床医院骨科,山西太原030001 [3]桂林医学院药学院,广西桂林541000

出　　处：《中国生化药物杂志》2015年第6期6-9,共4页Chinese Journal of Biochemical Pharmaceutics

基　　金：国家自然科学基金(81260462;81460474)

摘　　要：目的研究复方风湿宁(compound Fengshining,FSN)联合甲氨蝶呤(methotrexate,MTX)对类风湿性关节炎(rheumatoid arthritis,RA)滑膜成纤维细胞(fibroblast-like synoviocytes,FLS)Treg/Th17免疫平衡的影响。方法体外培养RA FLS,分别用FSN和MTX处理,分为FSN 1、2、3组(0.01、0.1、1.0 mg/m L FSN),MTX 1、2、3组(0.01、0.1、1.0 mg/m L MTX),联合1、2、3组(0.01 mg/m L FSN+0.01 mg/m L MTX、0.1 mg/m L FSN+0.1 mg/m L MTX、1.0 mg/m L FSN+1.0 mg/m L MTX),并以未加药的培养基作为对照组,共10组。采用MTT法检测各组RA FLS增殖情况,Real-time PCR法检测各组IL-10、IL-17、Foxp3、ROR-γt mRNA表达,ELISA检测各组IL-10、IL-17、IL-22水平。结果 FSN各组、MTX各组和联合各组FLS增殖OD值均低于对照组,尤以联合2组降低最显著(P<0.05)。FSN2组、MTX2组和联合2组IL-10 mRNA、Foxp3 mRNA表达均高于对照组,ROR-γt mRNA表达低于对照组,尤以联合2组变化显著(P<0.05)。FSN2组和联合2组IL-17mRNA表达均低于对照组,以联合2组降低显著(P<0.05)。FSN2组、MTX2组和联合2组IL-10水平均高于对照组、IL-22水平均低于对照组,以联合2组变化明显(P<0.05)。联合2组IL-17水平低于对照组(P<0.05)。结论 FSN联合MTX能够抑制RA FLS增殖,升高IL-10、Foxp3表达,降低IL-17、ROR-γt表达,调节Treg/Th17免疫平衡,以0.1 mg/m LFSN+0.1 mg/m L MTX的效应最为明显。Objective To study the effect of compound Fengshining （FSN） combined with methotrexate （MTX） on Treg/Thl7 immune balance in fibroblast-like synoviocytes （FLS） with rheumatoid arthritis （RA）. Methods RA FLS were cultured in vitro and treated with FSN and MTX. They were divided into ten groups： FSN 1, 2, 3 groups （0. 01, 0. 1, 1.0 mg/mL FSN）, MTX 1,2,3 groups （0.01,0. 1, 2. 0 mg/mL MTX）, combination 1,2, 3 groups （0. 01 mg/mL FSN ＋0. 01 mg/mL MTX, 0. 1 mg/mL FSN ＋0. 1 mg/mL MTX, 1.0 mg/mL FSN ＋ 1.0 mg/mL MTX） and the medium without drugs as control group. FLS proliferation was detected by MTY method. IL-10, IL-17, Foxp3, ROR--yt mRNA were detected by real-time PCR method. IL-10, IL-17, IL-22 levels were detected by ELISA method. Results OD values of FLS proliferation in FSN groups, MTX groups and combination groups were significantly lower than those in control group, especially combination 2 group changed significantly （ P 〈 0. 05 ）. IL-10 mRNA, Foxp3 mRNA in FSN 2 group, MTX 2 group and combination 2 group were higher and ROR-γt mRNA were lower than those in control group, especially combination 2 group changed significantly （P 〈0. 05）. IL-17 mRNA in FSN 2 group and combination 2 group were lower than those in control group, especially the combination 2 group changed significantly （ P 〈 0. 05 ）. IL-10 level in FSN 2 group, MTX 2 group and combination 2 group were higher and IL-22 levels were lower than those in control group, especially combination 2 group changed significantly （P 〈 0. 05）. IL-17 level in combination 2 group were lower than those in control group （P 〈 0.05）. Oonclusion Compound Fengshining combined with methotrexate could inhibit RA FLS proliferation, elevate IL-10 and Foxp3 expression, reduce the IL-17 and ROR-γt expression and adjust Treg/Thl7 immune balance; effect of 0. 1 mg/mL FSN ＋0. 1 mg/mLMTX is the most obvious.

关 键 词：甲氨蝶呤 复方风湿宁 类风湿性关节炎 调节性T细胞 

分 类 号：R593.2[医药卫生—内科学]