苗药金乌健骨方对类风湿关节炎滑膜细胞IL-17/IL-17R的影响  

Effects of Jin Wu Jiangu Recipe on IL-17/IL-17R in Rheumatoid Arthritis Synoviocytes

作　　者：李蓉[1] 姚血明[1] 宁乔怡 黄颖[1] 唐芳[1] 陆道敏[1] 徐晖[1] 马武开[1] LI Rong;YAO Xueming;NING Qiaoyi;HUANG Ying;TANG Fang;LU Daomin;XU Hui;MA Wukai(Department of Rheumatology and Immunology,the Second Affiliated Hospital of Guiyang of TCM,Guiyang 550003 Guizhou,China)

机构地区：[1]贵阳中医学院第二附属医院风湿免疫科,贵州贵阳550003

出　　处：《中药新药与临床药理》2018年第5期546-551,共6页Traditional Chinese Drug Research and Clinical Pharmacology

基　　金：国家自然科学基金项目(81360580);贵州省高层次创新型人才培养计划--"百"层次人才项目(黔科合平台人才[2016]5650);贵州省民族医学特色重点学科项目(黔学位合字ZDXK[2015]7号)

摘　　要：目的探讨苗药金乌健骨方总提物对类风湿关节炎滑膜组织成纤维样滑膜细胞(RA-FLS)白细胞介素6(IL-6)、转化生长因子-β(TGF-β)、白细胞介素17(IL^(-1)7)分泌及IL^(-1)7受体(IL^(-1)7R)蛋白的影响。方法采用组织块法体外培养原代RA-FLS;分为金乌健骨方低、中、高剂量组(0.06,0.6,6.0 mg·m L^(-1)),雷公藤多苷组(0.03 mg·m L^(-1))及空白对照组,共5组。采用ELISA法检测IL-6、TGF-β、IL^(-1)7含量;PCR法检测IL^(-1)7R m RNA表达;Western Blot法检测IL^(-1)7R蛋白表达。结果与空白对照组比较,各给药组细胞培养上清液中IL-6、TGF-β、IL^(-1)7的浓度均有不同程度的降低(P<0.01),IL^(-1)7R m RNA及蛋白表达水平均明显下降(P<0.01)。与雷公藤多苷组比较,金乌健骨方高剂量组IL-6、TGF-β、IL^(-1)7水平及IL^(-1)7R蛋白表达量明显降低(P<0.05),金乌健骨方中、高剂量组IL^(-1)7R m RNA表达水平明显降低(P<0.05)。结论苗药金乌健骨方总提物能够下调RA-FLS中IL-6、TGF-β、IL^(-1)7的分泌,降低IL^(-1)7R m RNA及蛋白表达量。Objective To investigate the effects of Jin Wu Jiangu recipe total extract on the inflammatory factors IL-6,TGF-β,IL^-17 and receptor protein IL^-17R in rheumatoid arthritis synovial fibroblasts（RA-FLS）.Methods RASF were cultivated in vitro by tissue piece method.RASF were divided into blank control group,Jin Wu Jiangu recipe low dose group,Jin Wu Jiangu recipe medium dose group,Jin Wu Jiangu recipe high dose group,and tripterygium glycosides control group,and treated with serum free medium,different doses of Jin Wu Jiangu recipe total extract（0.06 mg·mL^-1,0.6 mg·mL^-1,6.0 mg·mL^-1）,and tripterygium glycosides（0.03 mg·mL^-1） respectively for 24 h.The secretion of IL-6,TGF-β,IL-17 in cell culture supernatant were tested by ELISA,the gene expression of IL-17R mRNA were detected by PCR,the expression level of IL-17R protein were measured by Western Blot.Results Compared with blank control group,the concentration of IL-6,TGF-β,IL-17 in cell culture supernatant of each group were reduced to some extent,and had statistical differences（P〈0.01）;the high does group’s impact was better than the low,medium dose groups and tripterygium glycosides group（P〈0.01）.Both results from PCR and Western Blot displayed that,compared with blank control group,different doses of Jin Wu Jiangu recipe total extract and tripterygium glycosides had inhibitory effect on IL-17R at gene and protein level（P〈0.01）,the inhibitory effect in Jin Wu Jiangu recipe high dose group was obviously superior to low,medium dose groups and tripterygium glycosides group（P〈0.01）.Conclusion Miao medicine Jin Wu Jiangu recipe total extract can down-regulate the secretion of IL-6,TGF-β,IL-17,and reduce IL-17R at gene and protein level in RASF.

关 键 词：金乌健骨方 类风湿关节炎 滑膜细胞 白介素17 白介素17受体 

分 类 号：R285.5[医药卫生—中药学]