断藤益母汤对胶原诱导性关节炎大鼠软骨破坏的作用及机制研究  

Duanteng Yimu Decoction(断藤益母汤)Reduces Catabolism in Chondrocytes Co-Cultured with RA FLS by Regulating TGF-β Signaling Pathway

作　　者：韩隆胤 钱凯 杜彦仪 黄文广 曾丽盈 魏赈权 王强[1] 史周薇 李楠 林昌松[4] HAN Longyin;QIAN Kai;DU Yanyi;HUANG Wenguang;ZENG Liying;WEI Zhenquan;WANG Qiang;SHI Zhouwei;LI Nan;LIN Changsong(Guangzhou University of Chinese Medicine,Guangzhou 510405,Guangdong,China;Fangshan Hospital of Beijing University of Chinese Medicine,Beijing 102400,China;School of Traditional Chinese Medicine,Jinan University,Guangzhou 510632,Guangdong,China;The First Affiliated Hospital of Guangzhou University of Chinese Medicine,Guangzhou 510405,Guangdong,China)

机构地区：[1]广州中医药大学,广东广州510405 [2]北京中医药大学房山医院,北京102400 [3]暨南大学中医学院,广东广州510632 [4]广州中医药大学第一附属医院,广东广州510405

出　　处：《中华中医药学刊》2021年第7期107-111,I0022-I0026,共10页Chinese Archives of Traditional Chinese Medicine

基　　金：国家自然科学基金(81573930,81774262);中央高校基本科研业务费专项资金(21618335);广东省自然科学基金(2017A030311009,2018A0303130112);广东省医学科学技术研究基金(A2019066);广东省中医药局科研项目(20201072)。

摘　　要：目的研究断藤益母汤在胶原诱导性关节炎(collagen-induced arthritis, CIA)大鼠软骨破坏中的作用及其调节机制。方法取SPF级Wistar大鼠36只,随机分组为正常组、模型组、断藤益母汤低剂量组、断藤益母汤中剂量组、断藤益母汤高剂量组、甲氨蝶呤组(methotrexate, MTX)6组,除正常组外,其余5组大鼠采用二次免疫法构建CIA模型,二次免疫后当天开始连续给药28 d。观察并记录各组大鼠足肿胀度变化;HE染色法观察踝关节病理改变;番红固绿染色法观察软骨破坏;免疫组织化学法检测关节软骨中TGF-β1的表达;ELISA检测血清中MMP-9、MMP-13、TGF-β1表达。结果足肿胀度检测结果显示药物组大鼠足肿胀度较模型组减轻(P<0.01);HE染色显示断藤益母汤药物组及MTX组大鼠病理破坏对比模型组减轻;番红固绿染色结果显示断藤益母汤高剂量组与MTX组大鼠软骨破坏对比模型组明显较轻;免疫组化结果显示断藤益母汤药物组及MTX组大鼠软骨中TGF-β1蛋白表达减少(P<0.01);ELISA检测显示断藤益母汤与MTX降低了大鼠血清中MMP-9、MMP-13、TGF-β1的表达(P<0.05或P<0.01)。结论断藤益母汤能减轻CIA大鼠的关节肿胀程度和关节软骨破坏,与抑制MMP-9、MMP-13及TGF-β1的表达有关。Objective To investigate the effect of Duanteng Yimu Decoction(断藤益母汤,DTYMD) in the destruction of cartilage in collagen-induced arthritis(CIA) rats and its regulating mechanism. Methods Thirty-six Wistar rats of SPF grade were randomly divided into normal group, model group, DTYMD low-dose group, DTYMD middle-dose group, DTYMD high-dose group, and methotrexate(MTX) group. Except the normal group, the other 5 groups were established a collagen-induced arthritis model by using a secondary immunization method. On the day after the second immunization, continuous administration was started for 28 days. The changes in foot swelling of each group were observed and recorded. HE staining was used to observe pathological changes in the ankle joint. The saffron solid green staining was used to observe cartilage destruction. The immunohistochemical method was used to detect expression of TGF-β1 in articular cartilage. ELISA was used to detect expressions of MMP-9, MMP-13 and TGF-β1 in serum. Results Foot swelling test results showed that the swelling degree of the rats in the drug groups was reduced compared with that of the model group(P<0.01). The HE staining showed that the pathological damage of the rats in the DTYMD groups and the MTX group was alleviated. Saffron solid green staining showed that the cartilage destruction of rats in the DTYMD high-dose group and MTX group was significantly lighter. Immunohistochemical results showed that the expressions of TGF-β1 protein in the cartilage of rats in the DTYMD groups and MTX group were decreased(P<0.01). ELISA test showed that DTYMD and MTX reduced the expressions of MMP-9, MMP-13 and TGF-β1 in serum(P<0.05 or P<0.01).Conclusion DTYMD can reduce joint swelling and articular cartilage destruction in CIA rats, and it is related to inhibiting the expressions of MMP-9, MMP-13 and TGF-β1.

关 键 词：断藤益母汤 胶原诱导性关节炎 软骨 TGF-Β1 基质金属蛋白酶 

分 类 号：R285.5[医药卫生—中药学]