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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5 821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。
Abstract：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 type Ⅱ collagen 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(β-catenin) 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 5 821 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 M-puerarin intervention decreased(P<0.05). Conclusion Puerarin has the characteristic of multi-components, multi-targets and multi-pathway intervention in RA. Puerarin may alleviate synovial hyperplasia, reduce articular cartilage erosion and bone destruction in CIA in rats by inhibiting Wnt/β-catenin signaling pathway.
[1] Firestein G S,Budd R C,Gabriel S E,等.凯利风湿病学[M].9版.栗占国译.北京：北京大学医学出版社，2015:1146-57.[1] Firestein G S,Budd R C,Gabriel S E,et al.Kelley’s textbook of rheumatology[M].9 ed.Li Zhanguo Transl.Beijing:Peking University Medical Press,2015,1146-57.
[2] Chemin K,Gerstner C,Malmstr?m V.Effector functions of CD4+ T cells at the site of local autoimmune inflammation-lessons from rheumatoid arthritis[J].Front Immunol,2019,10:353.doi:10.3389/fimmu.2019.00353.
[3] 李娟，蒋扬青，沈瑞明，等.METTL3通过mRNA m6A甲基化促进类风湿关节炎滑膜成纤维细胞增殖、迁移及分泌炎症因子[J].安徽医科大学学报，2024,59(4):619-26.doi:10.19405/j.cnki.issn1000-1492.2024.04.010.[3] Li J,Jiang Y Q,Shen R M,et al.METTL3 promotes proliferation,migration,and secretion of inflammatory factors by mRNA m6A methylation in rheumatoid arthritis synovial fibroblasts[J].Acta Univ Med Anhui,2024,59(4):619-26.doi:10.19405/j.cnki.issn1000-1492.2024.04.010.
[4] Zhang B,Li M,Wang Q,et al.Exploring adverse effects of puerarin on catalase by multiple spectroscopic investigations and docking studies in vitro[J].J Biochem Mol Toxicol,2019,33(5):e22296.doi:10.1002/jbt.22296.
[5] Wang S,Zhang S,Wang S,et al.A comprehensive review on Pueraria:insights on its chemistry and medicinal value[J].Biomedecine Pharmacother,2020,131:110734.doi:10.1016/j.biopha.2020.110734.
[6] 朱卫丰，李佳莉，孟晓伟，等.葛属植物的化学成分及药理活性研究进展[J].中国中药杂志，2021,46(6):1311-31.doi:10.19540/j.cnki.cjcmm.20201124.601.[6] Zhu W F,Li J L,Meng X W,et al.Research advances in chemical constituents and pharmacological activities of Pueraria genus[J].China J Chin Mater Med,2021,46(6):1311-31.doi:10.19540/j.cnki.cjcmm.20201124.601.
[7] 曹盼，张樱山，魏学明，等.葛根素药理作用研究新进展[J].中成药，2021,43(8):2130-4.doi:10.3969/j.issn.1001-1528.2021.08.028.[7] Cao P,Zhang Y S,Wei X M,et al.New progress in pharmacological action of puerarin[J].Chin Tradit Pat Med,2021,43(8):2130-4.doi:10.3969/j.issn.1001-1528.2021.08.028.
[8] Wang D,Bu T,Li Y,et al.Pharmacological activity,pharmacokinetics,and clinical research progress of puerarin[J].Antioxidants,2022,11(11):2121.doi:10.3390/antiox11112121.
[9] 李凯，秦荣，邵佳乐，等.葛根素对废用性骨质疏松大鼠模型的防治作用及机制研究[J].中国中药杂志，2019,44(3):535-40.doi:10.19540/j.cnki.cjcmm.20181012.003.[9] Li K,Qin R,Shao J L,et al.Preventive effect and mechanism of puerarin on rat models of disuse osteoporosis[J].China J Chin Mater Med,2019,44(3):535-40.doi:10.19540/j.cnki.cjcmm.20181012.003.
[10] Liu F Y,Wang M Q,Liu M M,et al.Therapeutic effects of shikonin on adjuvant-induced arthritis in rats and cellular inflammation,migration and invasion of rheumatoid fibroblast-like synoviocytes via blocking the activation of Wnt/β-catenin pathway[J].Phytomedicine,2023,116:154857.doi:10.1016/j.phymed.2023.154857.
[11] Zhang Z,Chen X,Gao B,et al.PLD1 knockdown reduces metastasis and inflammation of fibroblast-like synoviocytes in rheumatoid arthritis by modulating NF-κB and Wnt/β-catenin pathways[J].Autoimmunity,2021,54(7):398-405.doi:10.1080/08916934.2021.1963957.
[12] Cai L,Mu Y R,Liu M M,et al.Penta-acetyl geniposide suppresses migration,invasion,and inflammation of TNF-α-stimulated rheumatoid arthritis fibroblast-like synoviocytes involving Wnt/β-catenin signaling pathway[J].Inflammation,2021,44(6):2232-45.doi:10.1007/s10753-021-01495-y.
[13] 李泽桦，何颖妍，曾宇宏，等.葛根素在大鼠体内的药代动力学特征和安全性的研究[J].中国临床药理学杂志，2023,39(10):1480-4.doi:10.13699/j.cnki.1001-6821.2023.10.024.[13] Li Z H,He Y Y,Zeng Y H,et al.Study on pharmacokinetic characteristics and safety of puerarin in rats[J].Chin J Clin Pharmacol,2023,39(10):1480-4.doi:10.13699/j.cnki.1001-6821.2023.10.024.
[14] Luo C F,Yuan M,Chen M S,et al.Pharmacokinetics,tissue distribution and relative bioavailability of puerarin solid lipid nanoparticles following oral administration[J].Int J Pharm,2011,410(1-2):138-44.doi:10.1016/j.ijpharm.2011.02.064.
基本信息:
DOI：10.19405/j.cnki.issn1000-1492.2025.01.004
中图分类号:R285.5
引用信息:
[1]高月,唐芳,马武开,等.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[J].安徽医科大学学报,2025,60(01):22-31.DOI:10.19405/j.cnki.issn1000-1492.2025.01.004.
基金信息:
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)~~