通痹合剂乙酸乙酯部位调节CIA大鼠细胞免疫的作用机制研究

目的评价通痹合剂乙酸乙酯部位(TBHJ)对胶原诱导型关节炎(CIA)大鼠的影响,揭示其抗炎抗风湿的作用机制。方法牛Ⅱ型胶原混合弗氏完全佐剂于Wistar大鼠尾根部注射造模,随机分为模型组,甲氨蝶呤(MTX)组和TBHJ治疗组,容积法评价后肢肿胀度;给药后30 d,乳腺钼靶机大鼠四肢摄片,计分法评价各组大鼠X光片四肢96块骨侵蚀和100个关节间隙变化;处死动物,取后肢踝关节苏木素-伊红染色,观察中性粒细胞、淋巴细胞和浆细胞浸润,滑膜细胞增生和软骨破坏的情况;免疫组化法观察关节内IL-17和CD28表达;RT-PCR法比较CD28 mRNA表达。结果 CIA大鼠出现严重的关节炎症和破坏,TBHJ给药后7 d即可明显抑制CIA大鼠关节肿胀;给药后30 d,TBHJ和MTX可明显抑制大鼠关节肿胀和关节破坏,与模型组比较有统计学差异(P<0.01);TBHJ可明显抑制CIA大鼠中性粒细胞、淋巴细胞、浆细胞的浸润、滑膜增生和软骨破坏(P<0.05),MTX可抑制滑膜增生和软骨破坏;CIA大鼠IL-17,CD28 mRNA和蛋白表达明显增加,TBHJ可明显抑制IL-17和CD28。结论 TBHJ具有抑制关节免疫炎症和关节破坏的作用,其机制可能是通过抑制T细胞第二信号分子CD28的表达,减少IL-17的产生实现的。

Pharmacological Mechanisms of Ethyl Acetate Extract of Tongbi Mixture on Cellular Immunity of Collagen induced Arthritis in Rats

Objective To evaluate the effect of ethyl acetate extract of Chinese medicine Tongbi Mixture(TM)on collagen induced-arthritis(CIA) rats,and to explore its anti-rheumatism pharmacological mechanisms.Methods The mixture of Bovine type collagen II(CII) and Freund’s complete adjuvant were injected intradermally at the base of Wistar rat tail for the establishment of CIA model.After evaluating the swelling of the hind limbs,CIA model rats were randomly divided to physiological saline group,methotrexate(MTX)group and TM group.Four limbs of the rats were photographed by molybdenum target mammography after treatment for 30 days.The scores of erosion in 96 bones and joint spacing narrow(JSN) in 100 joints were evaluated according to the results of X-ray mammography.At the end of the experiment,the rats were killed,both ankles of hind limbs were stained by HE dying,and neutrophil and lymphocyte count,plasmacyte infiltration,hyperplasia of synoviocytes and destruction of cartilage were assessed.Expression levels of IL-17 and CD28 protein in the joints were assayed by immunohistochemical method.Results Severe inflammation and bone damage were presented in CIA rat joints.The joint inflammation of CIA was inhibited after treatment with TM for 7 days.After treatment for 30 days,the inflammation and destruction of joints in CIA rats were obviously relieved in MTX group and TM group,the differences being significant compared with the physiological saline group(P<0.05).Neutrophil and lymphocyte count,plasmacyte infiltration,hyperplasia of synoviocytes and destruction of cartilage were inhibited by TM treatment.MTX depressed the hyperplasia of synoviocytes and destruc tion of cartilage.The expression levels of IL-17 and CD28 mRNA and protein were decreased in CIA model rats,and TM depressed significantly the expression of IL-17 and CD28.Conclusion TM could relieve joint inflammation and destruction of CIA rats.Its possible anti-rheumatism mechanism is related with the inhibition of CD28 expression and IL-17 production in T cells.