新安除湿补气汤对佐剂性关节炎大鼠治疗作用研究

Study on the Therapeutic Effect of Xin’an Chushi Buqi Decoction(除湿补气汤)on Adjuvant Arthritis Rats

目的研究新安经典名方除湿补气汤(dehumidification and tonic decoction,DTD)对类风湿性关节炎大鼠的治疗作用及可能的作用机制。方法取雄性SD大鼠40只,随机分为正常组(Normal)、模型组(AIA)、甲氨蝶呤组(MTX)和DTD组,每组10只。除正常组外,其余各组均采用弗氏完全佐剂(freund’s complete adjuvant,CFA)建立类风湿性关节炎大鼠模型(AIA model),模型成功后第2天开始灌胃给药,MTX、DTD组分别给予相应剂量的药物,Normal组与AIA组以同体积生理盐水代替药物,连续用药30 d。观察大鼠体质量、关节炎评分、脏器指数的变化;HE染色法观察大鼠踝关节组织形态学变化;酶联免疫吸附(ELISA)法检测大鼠血清TNF-α、IL-1β水平;生化检测乙酰胆碱(ACH)含量;Western Blot法检测大鼠滑膜α7nAChR的变化;RT-PCR检测脾中α7nAChR的表达。结果与Normal组相比,AIA组在造模第18天体质量明显下降,炎症评分较高,各脏器体积变小,大鼠关节出现明显组织破坏和炎性浸润,血清TNF-α、IL-1β水平明显升高,ACH含量较低,大鼠滑膜和脾中α7nAChR表达均明显降低;与AIA组相比,DTD组大鼠体质量明显上升,急性期大鼠关节肿胀度明显减轻,缓解期关节肿胀度被明显抑制,大鼠关节组织病理学变化得到显著改善,血清TNF-α、IL-1β水平明显降低,血清中ACH含量较高,大鼠滑膜和脾中α7nAChR表达均明显升高。结论DTD对佐剂性关节炎大鼠具有明显的治疗作用,其作用机制可能与其活化α7nAChR,激活胆碱能抗炎通路(CAP),升高体内ACH水平有关。

Objective To study the effect of Xin’an Chushi Buqi Decoction(除湿补气汤,CBD)on the treatment of rheumatoid arthritis in rats and its possible mechanism.Methods 40 male SD rats were randomly divided into normal group,model adjuvant induce arthritis(AIA)group,methotrexate(MTX)group and CBD group,with 10 rats in each group.Except the normal group,the rest of the groups adopted Freund’s complete adjuvant(CFA)established AIA model rat model,2 days after the success of the model mice,CBD,MTX group were given corresponding dose of the drug,normal group and the AIA group were treated with saline of the same volume for 30 consecutive days.The changes of body weight,arthritis score and organ index of rats were observed.HE staining was used to observe the morphological changes of the ankle joint.Serum levels of TNF-αand IL-1βwere determined by enzyme-linked immunosorbent assay.Biochemical detection of acetylcholine(ACH)content;The changes ofα7 nAChR in rat synovium were detected by Western Blot.The expression ofα7 nAChR in spleen was detected by Q-PCR.Results Compared with the normal group,the body weight of the AIA group decreased significantly on the 18 th day,the inflammatory score was higher,the organ index was lower,the serum TNF-αand IL-1βlevels were significantly increased,and the content of ACH was lower.The expression ofα7 nAChR in the synovium of the rats was lower detected by Western Blot,and the expression ofα7 nAChR in the spleen was down-regulated by PCR.Compared with the AIA group,the body weight of rats in the CBD group increased significantly,the degree of joint swelling in the acute stage was significantly reduced,and the degree of joint swelling in the remission stage was significantly inhibited.The organ coefficient was similar to that of normal rats.The serum levels of TNF-αand IL-1βdecreased significantly,and the serum content of ACH was higher.The articular histopathological changes of rats were significantly improved.The expression ofα7 nAChR in the synovium of rats was up-regulated by Western Blot.The expression ofα7 nAChR in spleen was up-regulated by RT-PCR.Conclusion CBD has obvious therapeutic effect on adjuvant arthritis in rats,and the mechanism of action may be related to the cholinergic anti-inflammatory pathway(CAP).