新风胶囊改善佐剂关节炎大鼠肺功能及调节T细胞功能的实验研究

Study of Xinfeng Capsule on the pulmonary function, regulatory T cells in the rats with adjuvant arthritis

目的:观察新风胶囊(XFC)对佐剂关节炎(AA)大鼠肺功能及调节T细胞(Treg)的影响。方法:将大鼠均分为正常对照(NC)组、模型对照(MC)组、甲氨喋呤(MTX)组、雷公藤多苷片(TPT)组和XFC低中高剂量(XFC-L、XFC-M、XFC-H)组,除NC组外,其余大鼠注射弗氏完全佐剂致炎,致炎后19天给药,NC、MC组给予0.9%氯化钠溶液,其余组分别给予MTX、TPT、XFC。用药30d后,采用肺功能仪检测大鼠肺功能,酶联免疫吸附(ELISA)法检测肿瘤坏死因子-α(TNF-α)、白介素-10(IL-10)表达,流式细胞术检测外周血CD4+CD2+5Treg表达,RT-PCR和免疫印迹(Western Blot)法检测Foxp3基因和蛋白表达。结果:MC组大鼠足跖肿胀度(E)、TNF-α升高;肺功能、IL-10、CD4+CD2+5Treg、Foxp3降低;各治疗组大鼠肺功能参数及IL-10、Treg、Foxp3的表达升高,E、TNF-α降低;XFC-M组变化较其他治疗组明显(P<0.05,P<0.01)。结论:AA大鼠存在肺功能降低,新风胶囊能改善AA大鼠关节症状及肺功能,其机制是通过上调IL-10、Treg及Foxp3的表达,下调TNF-α的表达,降低免疫复合物的沉积,减少炎症反应对组织的损伤,从而提高肺功能。

Objective： To observe the effects of Xinfeng Capsule（XFC） on pulmonary function and regulatory T cells（Treg） in adjuvant arthritis（AA） rats.Methods： Rats were averagely divided into normal control（NC）,model control（MC）,methotrexate（MTX）,Tripterygium glycosides tablet（TPT） and the low,medium,high dose of XFC（XFC-L,XFC-M,XFC-H） group.Excepting for the rats of normal group,the others were intracutaneously injected with Freund’s complete adjuvant.Administration in 19th day of proinflammatory.NC group and MC group were treated with 0.9% physiological saline.XFC group,MTX group and TPT group were treated with XFC,MTX,TPT respectively.Pulmonary function testing by spirometer,tumor necrosis factor α（TNF-α）,interleukin-10（IL-10） testing byenzyme-linked immunosorbent assay（ELISA）,expression of Treg by flow cytometry,Foxp3 gene and proteintesting by RT-PCR and immunoblottingassay respectively.Results： The level of swelling degree（E）,AI,TNF-α were significantly increased in MC group,pulmonary function parameters,IL-10,CD 4 ＋ CD 25 ＋ Treg,Foxp3 were significantly decreased.Compared with the MC group,the pulmonary function parameters and the expression of IL-10,Treg,Foxp3 were increased.Compared with the MC group,the pulmonary function parameters and the expression of IL-10,Treg,Foxp3 were increased.E,TNF-α were decreased in each treatment group rats.Compared with other treatment groups,XFC-M group was obvious（P0.05,P0.01）.Conclusion： It have reduced pulmonary function in AA rats.XFC can inhibit joint symptoms and improve lung function significantly.The mechanism is probably associated with up-regulating the expression of IL-10,Treg and Foxp3,and down-regulating TNF-α,decreasing immune complex deposition,reducing inflammation of the tissue damage,thereby improving lung function.