桂枝芍药知母汤干预佐剂性关节炎大鼠MAPKs和AP-1途径的实验研究

Experimental study of Guizhi Shaoyao Zhimu Decoction on MAPKs and AP-1 pathway in adjuvant arthritis rats

目的探讨桂枝芍药知母汤(GSZMT)通过丝裂原活化蛋白激酶(Mitogen-activated protein kinase,MAPKs)、激活子蛋白-1(activator protein 1,AP-1)干预佐剂性关节炎(AA)大鼠作用机制的研究。方法酶联免疫吸附法(ELISA)法检测外周血与足跖软组织核因子κB(NF-κBp65)表达,反转录实时定量PCR法(RT-qPCR)检测MAPKs(ERK1,ERK2,JNK,P38)mRNA表达,HE染色检测踝关节病理,免疫组织化学(IHC)法及免疫组织荧光法(IHF)检测踝关节及周围组织AP-1(c-jun,c-fos)、沉默调节蛋白1(Sirt1)的表达。结果与正常对照组比较,模型对照组NF-κBp65、MAPKs、AP-1蛋白表达均显著增高(P<0.01),Sirt1表达显著降低(P<0.01);与模型对照组比较,GSZMT组NF-κBp65、MAPKs、、AP-1蛋白表达均显著降低(P<0.01),Sirt1表达显著增高(P<0.01)。结论GSZMT能有效降低AA大鼠NF-κBp65、MAPKs、AP-1等炎症蛋白的表达,改善踝关节及周围组织的炎症表现,其治疗作用机理可能是通过降低MAPKs和AP-1途径的炎症蛋白,促进Sirt1蛋白表达实现的。

Objective To investigate the mechanism of Guizhi Shaoyao Zhimu Decoction(GSZMT)on adjuvant arthritis(AA)rats through mitogen activated protein kinases(MAPKs)and activator protein 1(AP-1).Methods NF-κBp65 in peripheral blood and foot-plantar soft tissue was detected by ELISA method.The mRNA expression of MAPKs(ERK1,ERK2,JNK,p38)was detected by(RT-qPCR),ankle pathology was detected by HE staining,the expression of AP-1(c-Jun,c-fos)and SIRT1 were detected by IHC and IF methods.Results Compared with the normal control group,the expression of NF-κBbp65,MAPKs and AP-1 in model control group increased significantly(P<0.01),and the expression of SIRT1 decreased significant-ly(P<0.01)Compared with the model control group,the expression of NF-κbp65,MAPKs,AP-1 in GSZMT groups de-creased significantly(P<0.01),and SIRT1 increased significantly(P<0.01).Conclusion Guizhi shaoyao Zhimu Decoction could effectively reduce the expression of inflammatory proteins in AA rats,such as NF-κBp65、MAPKs、AP-1,improve the in-flammation of ankle joint and surrounding tissues.The therapeutic mechanism might be achieved by reducing the inflammatory proteins of MAPKs and AP-1 pathway and promoting the expression of SIRT1 protein.