摘要
目的:采用网络药理学联合分子动力学模拟及细胞实验,揭示地黄饮子治疗膝骨关节炎的核心活性成分、关键靶点及作用机制。方法:通过中药系统药理学分析平台(TCMSP)、中药分子机制生物信息学分析工具(BATMAN-TCM)数据库筛选地黄饮子活性成分及靶点,结合GeneCards等数据库获取膝骨关节炎疾病靶点,构建“中药-活性成分-交集靶点-疾病”网络并进行GO/KEGG富集分析;利用AutoDock Vina、GROMACS软件进行分子对接及分子动力学模拟,验证核心成分与靶点的结合稳定性;通过IL-1β诱导软骨细胞炎症模型,采用CCK-8、阿利新蓝染色及免疫荧光、qPCR及Western Blot等方法,验证地黄饮子及核心成分的护软骨作用。结果:筛选出地黄饮子活性成分123种,对应靶点483个,与膝骨关节炎疾病靶点交集获得关键靶点77个(含IL-6、AKT1、CASP3、HIF-1α等);GO/KEGG富集分析显示其主要调控TNF信号通路、HIF-1α信号通路、PI3K-Akt信号通路等,涉及炎症调控、细胞凋亡及软骨基质代谢等生物过程。分子对接及动力学模拟证实,松果菊苷与膝骨关节炎核心靶点HIF-1α、CASP3结合稳定(平均结合自由能分别为-153.595,-147.57kJ/mol),复合物构象稳定。细胞实验表明,地黄饮子(4μg/mL)及松果菊苷(16μmol/L)可显著上调COL2A1表达,下调HIF-1α、CASP3及MMP13表达,保护软骨细胞基质完整性,抑制炎症损伤与凋亡。结论:地黄饮子通过“多成分-多靶点-多通路”协同作用治疗膝骨关节炎,核心成分松果菊苷可通过抑制HIF-1α介导的炎症级联反应及CASP3依赖的软骨细胞凋亡,发挥护软骨作用,可为地黄饮子临床治疗膝骨关节炎提供理论支撑。
Objective:To reveal the core active components,key targets,and mechanism of action of Dihuang Yinzi decoction in treatment of knee osteoarthritis(KOA)by using network pharmacology combined with molecular dynamics simulation(MDS)and cell experiments.Methods:The active components and targets of Dihuang Yinzi decoction were screened using the TCMSP and BATMAN-TCM databases,and the disease targets of KOA were obtained from the GeneCards and other databases.A“Chinese medicine-active component-interaction targets-disease”network was constructed and GO/KEGG enrichment analysis was performed.Molecular docking and MDS were conducted using AutoDock Vina and GROMACS software to verify the binding stability between core components and targets.The chondroprotective effect of Dihuang Yinzi decoction and core components was verified through IL-1β-induced chondrocyte inflammation models using CCK-8,Alcian blue staining,immunofluorescence,qPCR,and Western Blot methods.Results:123 active components of Dihuang Yinzi decoction were screened out,corresponding to 483 targets.The intersection with KOA disease targets yielded 77 key targets(including IL-6,AKT1,CASP3,HIF-1α,etc.).GO/KEGG enrichment analysis showed that it mainly regulated TNF signaling pathway,the HIF-1αsignaling pathway,PI3K-Akt signaling pathway,etc.,involving biological processes such as inflammation regulation,cell apoptosis and cartilage matrix metabolism.Molecular docking and dynamics simulations confirmed that echinacoside stably binding binds to the core targets of KOA,HIF-1αand CASP3,with average binding free energy was-153.595 kJ/mol and-147.57 kJ/mol,respectively,and the complex conformation was stable.Cell experiments showed that Dihuang Yinzi decoction(4μg/mL)and echinacoside(16μmol/L)could significantly up-regulate COL2A1 expression,down-regulate HIF-1α,CASP3,and MMP13 expression,protect the integrity of chondrocyte matrix,and inhibit inflammation and apoptosis.Conclusion:Dihuang Yinzi decoction treats KOA through the“multi-component-multi-target-multi-pathway”synergistic effect.The core component echinacoside can exert chondroprotective effects by inhibiting HIF-1α-mediated inflammatory cascade reactions and CASP3-dependent chondrocyte apoptosis,providing theoretical support for the clinical treatment of KOA with Dihuang Yinzi decoction.
作者
刘钊
杨杰
杜文喜
LIU Zhao;YANG Jie;DU Wenxi(The First Affiliated Hospital of Zhejiang Chinese Medical University(Zhejiang Provincial Hospital of Traditional Chinese Medicine),Hangzhou 310006,China)
出处
《中国中医骨伤科杂志》
2026年第1期67-77,共11页
Chinese Journal of Traditional Medical Traumatology & Orthopedics
基金
国家中医药综合改革示范区中医药科技共建项目(GZY-KJS-ZJ-2026-113)
浙江中医药大学附属第一医院2022年附属医院科研专项资助项目(2022FSYYZQ08)
浙江省中医药科技计划项目(2026ZL0330)
浙江省卫生高层次人才创新人才项目(202148)。
关键词
地黄饮子
膝骨关节炎
网络药理学
软骨保护
炎症调控
Dihuang Yinzi decoction
knee osteoarthritis
network pharmacology
chondroprotection
inflammation regulation
