二妙散通过α7nAChR-JAK2/STAT3途径改善胶原诱导性关节炎大鼠炎症的免疫学机制  

Immunological mechanism of Ermiao Powder improving inflammation in rats with collagen-induced arthritis throughα7nAChR-JAK2/STAT3 pathway

作　　者：刘静淑 岳进茹 李玉彤 李振 LIU Jing-shu;YUE Jin-ru;LI Yu-tong;LI Zhen(College of Basic Medical Sciences,Shanxi University of Chinese Medicine,Jinzhong 030619,China;National Key Laboratory for Innovation and Transformation of Luobing Theory,Shijiazhuang 050035,China;Shanxi Bethune Hospital(Tongji Shanxi Hospital,Shanxi Academy of Medical Sciences),the 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,山西晋中030619 [2]络病理论创新转化全国重点实验室,河北石家庄050035 [3]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,山西太原030032

出　　处：《中国中药杂志》2024年第13期3591-3599,共9页China Journal of Chinese Materia Medica

基　　金：国家自然科学基金项目(81904034);山西省基础研究计划面上项目(202303021211172);山西省中医药管理局科研课题(2023ZYYC058);山西中医药大学优秀青年科学家培育项目(2021PY-QN-04);山西中医药大学博士启动基金项目(2020BK02);中国博士后科学基金面上项目(2022M722004)。

摘　　要：探讨二妙散通过α7烟碱型乙酰胆碱受体(α7nAchR)-Janus激酶2(JAK2)/信号转导及转录激活因子3(STAT3)途径改善类风湿关节炎大鼠炎症的免疫学机制。56只雌性Wistar大鼠随机分为正常(HG)组、胶原诱导性关节炎模型(CM)组、迷走神经切断(VA)组、假手术(SH)组、二妙散治疗模型(EM)组、二妙散治疗迷走神经切断(EV)组、二妙散治疗假手术(ES)组,每组8只。除HG组外,其他组均构建胶原诱导性关节炎(CIA)大鼠模型,造模成功后进行颈部单侧迷走神经切断术及假手术(仅分离迷走神经),术后7 d开始给药治疗,持续治疗35 d。记录大鼠体质量及关节情况,观察大鼠脾脏病理学变化,ELISA检测大鼠血清中白细胞介素-6(IL-6)、白细胞介素-1β(IL-1β)、肿瘤坏死因子-α(TNF-α)含量,qRT-PCR、Western blot和免疫组化检测脾脏中α7nAchR-JAK2/STAT3通路核心基因mRNA及蛋白表达。结果显示,CM组对比HG组、VA组对比CM组和SH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<0.05,P<0.01),踝关节明显肿胀伴畸形,脾脏淋巴小结显著增大,血清中IL-6、IL-1β、TNF-α含量显著增加(P<0.05,P<0.01),脾脏中JAK2、STAT3 mRNA水平显著升高(P<0.05,P<0.01),磷酸化JAK2(p-JAK2)/JAK2、磷酸化STAT3(p-STAT3)/STAT3水平显著升高(P<0.05,P<0.01),JAK2、p-JAK2、STAT3、p-STAT3细胞数增加(P<0.05,P<0.01)。EM组对比CM组、ES组对比SH组,大鼠体质量显著升高(P<0.01),关节炎评分显著下降(P<0.05,P<0.01),踝关节肿胀度明显减轻,脾脏淋巴小结体积减小,炎症减轻,血清中IL-6、IL-1β、TNF-α含量显著下降(P<0.05,P<0.01),脾脏JAK2、STAT3 mRNA表达显著降低(P<0.05,P<0.01),p-JAK2/JAK2、p-STAT3/STAT3水平显著降低(P<0.05,P<0.01),JAK2、p-JAK2、STAT3、p-STAT3细胞数量显著减少(P<0.05,P<0.01),α7nAChR mRNA及蛋白表达显著上升(P<0.01)。EV组对比VA组,大鼠体质量上升及关节炎评分无明显差异,脾脏淋巴小结数量减少不显著,体�This study investigated the immunological mechanisms of Ermiao powder in the treatment of rheumatoid arthritis rats through the alpha 7 nicotinic acetylcholine receptor(α7nAChR)-Janus kinases 2(JAK2)/signal transducer and activator of transcription 3(STAT3)signaling pathway.A total of 56 female Wistar rats were randomly divided into the normal group(HG,n=8),collagen-induced arthritis(CIA)model group(CM,n=8),vagotomy group(VA,n=8),sham group(SH,n=8),Ermiao Powder treatment model group(EM,n=8),Ermiao Powder treatment for vagotomy group(EV,n=8)and Ermiao Powder treatment for sham group(ES,n=8).Following the establishment of CIA models in all groups except the HG group,the rats underwent unilateral vagotomy and sham operation(only the vagus nerve was separated).Drug treatment was started 7 days after surgery and continued for 35 days.The body weight and joints of rats were recorded,the pathological changes of the spleen of rats were observed,the contents of interleukin-6(IL-6),interleukin-1β(IL-1β)and tumor necrosis factor-α(TNF-α)in serum were detected by enzyme-linked immunosorbent assay(ELISA),and the mRNA and protein expression ofα7nAChR-JAK2/STAT3 pathway core genes in spleen were detected by qRT-PCR,Western blot and immunohistochemistry.Results showed that CM group(compared with HG group)and VA group(compared with CM group and SH group)had significantly decreased body weight(P<0.05,P<0.01),increased arthritis score(P<0.05,P<0.01),swollen ankle joints with deformity,and increased and enlarged lymph nodes in the spleen.There were also notable increases in the serum levels of IL-6,IL-1βand TNF-α(P<0.05,P<0.01),and in the mRNA expressions of JAK2 and STAT3 in the spleen(P<0.05,P<0.01).The protein levels of phosphorylated JAK2(p-JAK2)/JAK2 and phospho-STAT3(p-STAT3)/STAT3 were significantly increased(P<0.05,P<0.01),and the number of JAK2,p-JAK2,STAT3 and p-STAT3 cells increased(P<0.05,P<0.01).EM group(compared with CM group)and ES group(compared with SH group)exhibited significantly increased body weight(P<0

关 键 词：α7nAChR-JAK2/STAT3 胆碱能抗炎通路 类风湿关节炎 迷走神经 二妙散 

分 类 号：R285.5[医药卫生—中药学]