基于网络药理学和动物实验探讨葛根素治疗类风湿关节炎的作用机制  

Mechanism of puerarin in treatment of rheumatoid arthritis based on network pharmacology and animal experimental verification

作　　者：高月 唐芳 马武开 兰维娅 蒋总 金泽旭 Gao Yue;Tang Fang;Ma Wukai;Lan Weiya;Jiang Zong;Jin Zexu(Second School of Clinical Medicine,Guizhou University of Chinese Medicine,Guiyang 550002;Dept of Internal Medicine,Gaoyou Hospital of Traditional Chinese Medicine,Gaoyou 225699;Dept of Rheumatology and Immunology,the Second Affiliated Hospital of Guizhou University of Chinese Medicine,Guiyang 550003)

机构地区：[1]贵州中医药大学第二临床医学院,贵阳550002 [2]高邮市中医医院内科,高邮225699 [3]贵州中医药大学第二附属医院风湿免疫科,贵阳550003

出　　处：《安徽医科大学学报》2025年第1期22-31,共10页Acta Universitatis Medicinalis Anhui

基　　金：国家自然科学基金(编号:82160917);贵州省科技计划项目(编号:黔科合基础-ZK[2023]一般435);贵州省高等学校重点实验室建设项目(编号:黔教技[2023]017号);贵州省中医药管理局中医药、民族医药科学技术研究项目(编号:QZYY-2020-005)。

摘　　要：目的通过网络药理学和动物实验探讨葛根素治疗类风湿关节炎(RA)的作用机制。方法使用中药系统药理学数据库和分析平台(TCMSP)和SwissTargetPrediction数据库收集葛根素靶点,使用GeneCards、OMIM数据库获取RA的疾病相关靶点,利用Cytoscape 3.7.2软件建立蛋白-蛋白相互作用(PPI)网络,通过Metascape数据库进行基因本体(GO)功能和京都基因与基因组百科全书(KEGG)富集分析;使用Ⅱ型胶原乳剂复制RA大鼠-胶原诱导型关节炎(CIA)模型,将49只Wistar大鼠随机分为空白对照(Con)组,CIA模型(CIA)组,低、中、高剂量葛根素(L-、M-、H-puerarin)组,甲氨蝶呤(MTX)组,雷公藤多苷片(TGT)组。除Con组外,其余各组大鼠制成CIA大鼠模型后连续灌胃28 d。观察各组大鼠后肢关节红肿情况及踝关节病理改变,Western blot检测滑膜糖原合成酶激酶3β(GSK-3β)、β-连环蛋白(β-catenin)蛋白表达,qPCR检测滑膜GSK-3β、β-catenin、c-Myc mRNA表达。结果获得134个葛根素作用靶点,RA疾病相关靶点基因5821个,葛根素与RA交集靶点基因102个,涉及JAK-STAT、NF-κB、Wnt等184条信号通路。动物实验结果表明,M-puerarin和MTX干预后大鼠后足红肿症状改善并且关节滑膜中炎性细胞浸润明显减少,软骨及骨组织破坏程度减轻。与CIA组相比,M-puerarin干预后的大鼠滑膜组织中GSK-3β、β-catenin蛋白表达和GSK-3β、β-catenin、c-Myc mRNA表达均下降(P<0.05)。结论葛根素可以通过多靶点、多途径协同治疗RA,可能通过抑制Wnt/β-catenin信号通路,缓解CIA大鼠滑膜增生,减轻关节软骨侵蚀及骨破坏情况。Objective To investigate the mechanism of puerarin in the treatment of rheumatoid arthritis(RA)by network pharmacology and animal experiments.Methods Traditional Chinese Medicine Systems Pharmcolog Database(TCMSP)and SwissTargetPrediction database were used to collect puerarin targets,and the targets of RA were obtained from GeneCards database and OMIM database.The PPI network was established by Cytoscape 3.7.2 software.Gene ontology(GO)function and Kyotoencyclopedia of genes(KEGG)enrichment analysis were performed through the Metascape database.RA rat-collagen-induced arthritis(CIA)model was reproduced using typeⅡcollagen emulsion,49 Wistar rats were randomly assigned to seven groups:control group,CIA model group,low-dose,medium-dose and high-dose puerarin group,methotrexate group,Tripterysium Glycosides Tablets group.Except for the control group,the other groups were given continuous gavage for 28 days after the CIA in rats model were prepared.The redness and swelling of the joints and ankle joint pathological changes were observed in each group.Western blot was used to detect the expression of Glycogen synthase kinase3β(GSK-3β),beta-catenin(βcatenin)proteins in the synovium.Real-time quantitative polymerase chain reaction(qPCR)was used to detect the expression of GSK-3β,β-catenin and c-Myc mRNA in the synovium.Results Puerarin had 134 targets genes,RA had 5821 target genes,and there were 102 overlapping target genes of puerarin and RA.It involved 184 signaling pathways,including JAK-STAT signaling pathway,NF-κB signaling pathway,Wnt signaling pathway,et al.The results of animal experiments showed that after the intervention of M-puerarin and MTX,the symptoms of redness and swelling of the hind foot were alleviated,the inflammatory cell infiltration in the synovium of the joint was significantly reduced,and the damage of cartilage and bone tissue was reduced.Compared with CIA group,the expressions of GSK-3β,β-catenin protein and GSK-3β,β-catenin and c-Myc mRNA in synovial tissue of rats after Mpuera

关 键 词：葛根素 类风湿关节炎 CIA大鼠 网络药理学 WNT/Β-CATENIN信号通路 滑膜增生 

分 类 号：R285.5[医药卫生—中药学]