经方二妙散通过ChAT/α7nAChR/NF-κB途径治疗胶原诱导性关节炎大鼠的作用机制研究  

Study on mechanism of Ermiao powder in treatment of collagen-induced arthritis rats through ChAT/α7 nAChR/NF-κB pathway

作　　者：李玉彤 刘静淑 李振 LI Yutong;LIU Jingshu;LI Zhen(College of Basic Medical Sciences,Shanxi University of Chinese Medicine,Jinzhong 030619,China;Shanxi Bethune Hospital(Shanxi Academy of Medical Sciences,Tongji Shanxi Hospi-tal,Third Hospital of Shanxi Medical University),Taiyuan 030032,China)

机构地区：[1]山西中医药大学基础医学院,晋中030619 [2]山西白求恩医院(山西医学科学院,同济山西医院,山西医科大学第三医院),太原030032

出　　处：《中国免疫学杂志》2025年第2期374-380,386,共8页Chinese Journal of Immunology

基　　金：国家自然科学基金青年科学基金(81904034);山西省基础研究计划面上项目(202303021211172);中国博士后科学基金(2022M722004);山西中医药大学博士启动基金(2020BK02);山西中医药大学优秀青年科学家培育项目(2021PY-QN-04);山西中医药大学研究生创新创业项目(2021CX015);基于炎性反应的重大疾病创新药物山西省重点实验室开放基金(2021sxcxyw01);山西省中医药管理局科研课题(2023ZYYC058)。

摘　　要：目的:探讨经方二妙散通过ChAT/α7nAChR/NF-κB途径治疗胶原诱导性关节炎(CIA)大鼠的作用机制。方法:56只SPF级雌性Wistar大鼠随机分为正常对照(NOR)组、CIA模型(CIA)组、迷走神经切断(VAO)组、假手术(SHO)组、二妙散治疗CIA大鼠(EMS)组、二妙散治疗迷走神经切断(EVAO)组和二妙散治疗假手术(ESHO)组。除NOR组外,其他组均构建CIA大鼠模型,颈部单侧迷走神经切断及假手术7 d后开始给药,持续35 d。记录大鼠体质量及关节情况,观察大鼠脾脏和关节组织病理形态,检测血清IL-1、IL-6、TNF-α含量及关节中ChAT/α7nAChR/NF-κB通路核心基因mRNA及蛋白表达,免疫组化定位分析。结果:CIA组对比NOR组/VAO组对比CIA组和SHO组,大鼠体质量显著降低(P<0.05,P<0.01),关节炎评分显著升高(P<0.01),踝关节明显肿胀(伴畸形),脾脏淋巴样白髓和主要生发中心增生明显,关节腔隙缩小(破坏)伴炎症细胞浸润,血清IL-1、IL-6、TNF-α含量显著增加(P<0.05,P<0.01),关节CHRNA7、ChAT、IκBα、NF-κB p50/p65 mRNA及蛋白水平显著升高,且主要表达于关节面(P<0.05,P<0.01)。EMS组对比CIA组/ESHO组对比SHO组,大鼠体质量显著升高,关节炎评分显著下降(P<0.05),踝关节肿胀度明显减轻,脾脏白髓和生发中心明显减少,关节面炎症细胞浸润减少,关节腔隙增大,血清IL-1、IL-6、TNF-α含量下降,CHRNA7、ChAT、IκBα mRNA及蛋白水平表达趋势上升,NF-κB p50/p65 mRNA及蛋白水平表达趋势下降,且主要位于关节面。EVAO组对比VAO组,大鼠体质量显著下降(P<0.05),血清IL-1含量显著增加(P<0.01),IL-6、TNF-α含量差异无统计学意义(P>0.05),CHRNA7、ChAT、IκBα mRNA及蛋白水平表达趋势下降,NF-κB p50/p65 mRNA及蛋白水平表达趋势上升,且主要表达于关节面。结论:经方二妙散可能通过ChAT介导α7nAChR高表达,从而下调NF-κB p50/p65,进而显著降低CIA大鼠炎症因子表达,缓解机体炎症和关节病变程度,最终有�Objective:To investigate therapeutic effect of Ermiao powder on collagen-induced arthritis(CIA)rats through ChAT/α7nAChR/NF-κB pathway.Methods:Fifty-six SPF female Wistar rats were randomly divided into normal control(NOR)group,CIA model(CIA)group,vagotomy(VAO)group,sham operation(SHO)group,Ermiao powder treat CIA rats(EMS)group,Ermiao powder treat vagotomy(EVAO)group and Ermiao powder treat sham operation(ESHO)group.CIA rats model was established in all groups except NOR group.Administration of drug began 7 days after unilateral vagotomy and sham operation,continued for 35 days.Body weight and joint condition of rats were recorded,and pathological morphology of spleen and joint tissues of rats were observed.Serum levels of IL-1,IL-6 and TNF-αwere detected,mRNA and protein expressions of ChAT/α7nAChR/NF-κB pathway core genes in joints were detected,and analyzed by immunohistochemical localization.Results:Compared with NOR/VAO group,body weight of rats in CIA group/SHO group was significantly decreased(P<0.05,P<0.01),arthritis score was significantly increased(P<0.01),ankle joint was obviously swollen(with deformity),lymphoid white pulp and main germinal center hyperplasia in spleen were obvious,joint lacuna reduction(destruction)with inflammatory cell infiltration,serum levels of IL-1,IL-6 and TNF-αwere significantly increased(P<0.05,P<0.01),mRNA and protein levels of CHRNA7,ChAT,IκBαand NF-κB p50/p65 were significantly increased and expressed in articular surface(P<0.05,P<0.01).Compared with CIA group/SHO group,body weight of rats in EMS group/ESHO group was significantly increased,arthritis score was significantly decreased(P<0.05),swelling degree of ankle joint was significantly reduced,white pulp and germinal center of spleen were significantly reduced,infiltration of inflammatory cells in articular surface was reduced,joint space was increased,serum levels of IL-1,IL-6 and TNF-αwere decreased,mRNA and protein expressions of CHRNA7,ChAT and IκBαwere increased,while mRNA and protein expressions of

关 键 词：胶原诱导性关节炎大鼠 类风湿关节炎 二妙散 ChAT/α7nAChR/NF-κB 胆碱能抗炎通路 迷走神经 

分 类 号：R392.9[医药卫生—免疫学]