中国组织工程研究

中国组织工程研究 ›› 2026, Vol. 30 ›› Issue (33): 8618-8630.doi: 10.12307/2026.285

• 骨组织构建 bone tissue construction • 上一篇    下一篇

淫羊藿治疗酒精性股骨头坏死：网络药理学及分子动力学模拟的作用机制

赖  渝1，2，陈跃平2，章晓云2，卓映宏2   

  1. 1广西中医药大学，广西壮族自治区南宁市   530001；2广西中医药大学附属瑞康医院，广西壮族自治区南宁市   530001
  • 收稿日期:2025-08-21 修回日期:2026-01-27 出版日期:2026-11-28 发布日期:2026-06-09
  • 通讯作者: 陈跃平，博士，主任医师，博士生导师，广西中医药大学附属瑞康医院，广西壮族自治区南宁市 530001
  • 作者简介:赖渝，男，1996年生，江西省赣州市人，汉族，博士，医师，主要从事四肢创伤、骨与关节方面研究。
  • 基金资助:
    广西壮族自治区临床重点专科(创伤外科)建设项目(桂卫医发〔2021〕17号)，项目负责人：陈跃平；广西中医药大学A类“桂派中医药传承创新团队”(2022A004)，项目负责人：陈跃平；广西中医药大学研究生教育创新计划项目(YCBXJ2023025)，项目负责人：赖渝；广西中医药大学青年基金项目(2020QN013)，项目负责人：卓映宏

Epimedium in the treatment of alcohol-induced osteonecrosis of the femoral head: action mechanism through network pharmacology and molecular dynamics simulation

Lai Yu1, 2, Chen Yueping2, Zhang Xiaoyun2, Zhuo Yinghong2   

  1. 1Guangxi University of Chinese Medicine, Nanning 530001, Guangxi Zhuang Autonomous Region, China; 2Ruikang Hospital, Guangxi University of Chinese Medicine, Nanning 530001, Guangxi Zhuang Autonomous Region, China
  • Received:2025-08-21 Revised:2026-01-27 Online:2026-11-28 Published:2026-06-09
  • Contact: Chen Yueping, PhD, Chief physician, Doctoral supervisor, Ruikang Hospital, Guangxi University of Chinese Medicine, Nanning 530001, Guangxi Zhuang Autonomous Region, China
  • About author:Lai Yu, PhD, Physician, Guangxi University of Chinese Medicine, Nanning 530001, Guangxi Zhuang Autonomous Region, China; Ruikang Hospital, Guangxi University of Chinese Medicine, Nanning 530001, Guangxi Zhuang Autonomous Region, China
  • Supported by:
    Clinical Key Specialty of Guangxi Zhuang Autonomous Region (Trauma Surgery) Construction Project, No. [2021]17 (to CYP); Guangxi University of Chinese Medicine Class A “Guangxi Traditional Chinese Medicine Inheritance and Innovation Team”, No. 2022A004 (to CYP); Graduate Education Innovation Program Project of Guangxi University of Chinese Medicine, No. YCBXJ2023025 (to LY); Youth Fund Project of Guangxi University of Chinese Medicine, No. 2020QN013 (to ZYH)

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摘要：


文题释义：
超高效液相色谱-四级杆-静电场轨道阱串联质谱(ultra-high performance liquid chromatography-Q exactive-mass spectrometry，UHPLC-QE-MS)：是一种集高效分离、高分辨质谱检测于一体的现代分析技术，广泛应用于天然产物成分鉴定、代谢产物分析、药物分析等领域。
分子动力学：是指通过计算机模拟技术，在给定温度和压力等条件下，研究原子和分子在一段时间内的运动轨迹和相互作用，从而探索其结构变化、动态行为和相互作用机制的科学方法。

背景：目前酒精性股骨头坏死的治疗及早期干预和延缓病情进展方面效果不理想。前期研究发现，淫羊藿的活性成分具有维持骨稳态平衡、促进骨折愈合的功效。
目的：结合网络药理学分析、分子对接技术、分子动力学模拟及体外细胞实验等多种方法，综合探讨淫羊藿干预酒精性股骨头坏死的潜在作用机制。
方法：采用超高效液相色谱-四级杆-静电场轨道阱串联质谱技术完成淫羊藿主要成分的鉴定工作后，首先基于TCMSP数据库对其潜在活性成分进行筛选，并联合TCMSP与Uniprot数据库获取相应的作用靶点；随后，从多个疾病数据库中提取与酒精性股骨头坏死相关的疾病靶点。将药物靶点与疾病靶点进行交集分析，将结果导入STRING平台构建蛋白质相互作用网络，并运用Cytoscape软件对网络拓扑结构进行解析，从中筛选出关键靶点。进一步构建“活性成分-靶点”关联网络，以确定核心活性成分。在此基础上，对筛选出的关键靶点进行GO功能注释和KEGG通路富集分析，并对核心成分与关键靶点进行分子对接研究，筛选出结合能最优的活性物质与受体开展分子动力学研究，以验证其稳定性和相互作用机制，最后对MC3T3-E1细胞(空白组、模型组、药物组)进行CCK-8及Western blot验证。
结果与结论：①研究共从淫羊藿提取液中鉴定出88种活性成分，通过成分靶点与酒精性股骨头坏死相关疾病靶点的交集分析，筛选出50个共同靶点，进一步确定5个核心活性成分及6个关键作用靶点。GO功能富集分析共涉及1 321个条目，KEGG通路分析识别出79条相关信号通路。②分子对接结果表明，核心成分与关键靶点之间具有较强的结合活性，其中淫羊藿苷与丝氨酸/苏氨酸蛋白激酶1的结合能最低；在分子动力学模拟中，淫羊藿苷与丝氨酸/苏氨酸蛋白激酶1之间的复合物结构表现出良好的稳定性。③体外实验证实，乙醇对MC3T3-E1成骨细胞具有明显毒性，抑制其成骨分化能力，而淫羊藿苷能够明显缓解该损伤效应，且该作用机制可能与丝氨酸/苏氨酸蛋白激酶信号通路的激活密切相关。通过系统分析淫羊藿的化学成分表明，其可能通过多组分协同作用、多靶点调控以及多途径介导的方式，在免疫调节、炎症抑制、氧化应激缓解、血管新生促进及骨代谢平衡等方面发挥治疗作用，从而有效改善酒精性股骨头坏死的病理进程。   
https://orcid.org/0000-0003-0172-7642 (赖渝) 


中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程

关键词: 淫羊藿, 酒精性股骨头坏死, 分子动力学模拟, 质谱, 分子对接, 网络药理学

Abstract: BACKGROUND: Currently, the treatment, early intervention, and delaying the progression of alcoholic osteonecrosis of the femoral head are not satisfactory. Preliminary studies have found that the active components of Epimedium have the effects on maintaining bone homeostasis and promoting fracture healing.
OBJECTIVE: To investigate the potential mechanism of Epimedium in the treatment of alcohol-induced osteonecrosis of the femoral head by integrated approaches, including network pharmacology, molecular docking technology, molecular dynamics simulations, and in vitro cell experiments.
METHODS: After identifying the main constituents of Epimedium via ultra-high performance liquid chromatography coupled with quadrupole-exactive Orbitrap high-resolution mass spectrometry, potential active compounds were screened firstly based on the TCMSP database, and their corresponding targets were retrieved using the TCMSP and Uniprot databases. Subsequently, disease-related targets associated with alcohol-induced osteonecrosis of the femoral head were extracted from multiple disease databases. An intersection analysis was conducted between drug targets with disease targets, and the results were imported into the STRING platform to construct a protein-protein interaction network. The network topology was analyzed using Cytoscape software to identify key targets. An “active ingredient–target” interaction network was then constructed to determine core active compounds. On this basis, Gene Ontology annotation and Kyoto Encyclopedia of Genes and Genomes pathway enrichment analyses were performed on the key targets. Molecular docking was conducted between core compounds and key targets. The most stable compound-target complex, based on binding affinity, was further subjected to molecular dynamics simulations to validate its structural stability and interaction mechanisms. Finally, cell counting kit-8 and western blot assays were performed on MC3T3-E1 cells in the blank group, model group and drug group.
RESULTS AND CONCLUSION: (1) A total of 88 active compounds were identified from Epimedium extract. Fifty overlapping targets were obtained by intersecting the compound-related and disease-related targets, from which five core compounds and six key targets were identified. Gene Ontology enrichment involved 1 321 terms, and Kyoto Encyclopedia of Genes and Genomes analysis revealed 79 signaling pathways. (2) Molecular docking showed strong binding affinities between the core compounds and key targets, with icariin exhibiting the lowest binding energy to serine/threonine protein kinase 1. Subsequent molecular dynamics simulations demonstrated good structural stability between icariin and serine/threonine protein kinase 1. (3) In vitro experiments confirmed that alcohol exerted significant cytotoxic effects on MC3T3-E1 osteoblasts, impairing their osteogenic differentiation, while icariin effectively alleviated these detrimental effects, potentially through activation of the serine/threonine protein kinase signaling pathway. A systematic analysis of the chemical constituents of Epimedium suggests that it may exert therapeutic effects against alcohol-induced osteonecrosis of the femoral head via multi-component synergy, multi-target regulation, and multi-pathway modulation. These mechanisms may involve immune regulation, anti-inflammatory activity, mitigation of oxidative stress, promotion of angiogenesis, and maintenance of bone metabolic homeostasis, ultimately contributing to the improvement of alcohol-induced osteonecrosis of the femoral head pathological progression.

Key words: Epimedium, alcohol-induced osteonecrosis of the femoral head, molecular dynamics simulation, mass spectrometry, molecular docking, network pharmacology

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