整合网络药理学、分子对接和分子动力学探讨甘麦大枣汤治疗失眠的作用机制

Potential mechanisms of Ganmai Dazao Decoction for treating insomnia： an integration of network pharmacology， molecular docking， and molecular dynamics simulation

中文关键词:  甘麦大枣汤  失眠  网络药理学  分子对接  分子动力学

英文关键词:Ganmai Dazao Decoction  Insomnia  Network pharmacology  Molecular docking  Molecular dynamics

中文摘要:

      背景 失眠是一种常见的睡眠障碍，与心血管疾病、糖尿病及心理健康密切相关，严重影响个体的生活质量。临床常用药物治疗虽有效，但长期使用易引发耐药性和依赖性。甘麦大枣汤改善失眠的效果确切，但其分子机制尚不清楚。目的 探讨甘麦大枣汤治疗失眠的活性成分及核心靶点，系统揭示其潜在的分子药理机制，为临床应用提供参考。方法 于2024年11月，从INPUT数据库筛选甘麦大枣汤的活性成分及相关靶点，利用基因表达综合（GEO）数据库，获取与失眠相关的数据集，使用GEO2R工具进行差异分析，获取失眠的差异表达基因（DEGs）。通过韦恩图获取共有靶点，采用STRING数据库和Cytoscape 3.9.1构建蛋白质互作（PPI）网络。对共有靶点进行基因本体（GO）和京都基因与基因组百科全书（KEGG）富集分析。选取节点度值排名前3位的关键活性成分和前10位核心靶点，使用AutoDock 4.4.6对受体和配体进行分子对接以及分子动力学模拟，并采用Pymol 3.0.3将结果可视化，进一步验证受体-配体复合物体系的稳定性。结果 检索出甘麦大枣汤中的337种活性成分及5 265个药物相关靶点，共收集到1 061个失眠的DEGs。甘麦大枣汤与失眠的共有靶点共287个。中药-活性成分-共有靶点-疾病网络显示，槲皮素、儿茶素和山柰酚是甘麦大枣汤治疗失眠的关键成分。这3种成分可作用于核因子κB抑制因子α（NFKBIA）、纤维连接蛋白1（FN1）、白细胞介素6（IL6）、蛋白质c-FOS（FOS）、腺病毒E1A结合蛋白P300（EP300）、组蛋白去乙酰化酶1（HDAC1）、转录因子JUN（JUN）、热休克蛋白90α（HSP90AA1）、甘油醛-3-磷酸脱氢酶（GAPDH）、白细胞介素1β（IL1β）共10个核心靶点，发挥治疗失眠的作用。GO和KEGG富集分析显示，甘麦大枣汤可能通过白细胞介素17（IL17）信号通路、脂质与动脉粥样硬化等信号通路发挥治疗失眠的作用。分子对接结果显示，甘麦大枣汤的3种关键成分与10个核心靶点均有良好的结合活性。分子动力学模拟显示，槲皮素-GAPDH、儿茶素-HDAC1和山奈酚-EP300复合体均能达到稳定状态。结论 甘麦大枣汤的关键成分槲皮素、儿茶素、山奈酚可作用于10个核心靶点，通过调控IL17信号通路、脂质和动脉粥样硬化通路等多种途径，发挥对失眠的治疗作用。

英文摘要:

      Background Insomnia， a common sleep disorder， is robustly associated with cardiovascular diseases， diabetes， and psychiatric disorders， substantially impairing quality of life. Although clinically commonly used medications are effective， long-term use may lead to drug resistance and dependence. While the efficacy of Ganmai Dazao Decoction in improving insomnia is definite， its underlying molecular mechanisms remain unclear.Objective To explore the active ingredients and core targets of Ganmai Dazao Decoction in the treatment of insomnia， systematically reveal its potential molecular pharmacological mechanism， and to provide references for clinical application.Methods In November 2024， the active ingredients and related targets of Ganmai Dazao Decoction were screened from the INPUT database. Insomnia-related datasets were acquired from the Gene Expression Omnibus （GEO） database， followed by differential expression analysis using GEO2R to identify differentially expressed genes （DEGs） associated with insomnia. The shared targets were obtained through Venn diagrams， and the protein-protein interaction （PPI） network was constructed using the STRING database and Cytoscape 3.9.1. Enrichment analyses were conducted on the shared targets using Gene Ontology （GO） and the Kyoto Encyclopedia of Genes and Genomes （KEGG）. The top 3 key active ingredients and the top 10 core targets in terms of node degree values were selected. Molecular docking and molecular dynamics simulation of receptors and ligands were performed using AutoDock 4.4.6， and the results were visualized using Pymol 3.0.3 to further verify the stability of the receptor-ligand complex system.Results A total of 337 active ingredients and 5 265 drug-related targets in Ganmai Dazao Decoction were retrieved， along with 1 061 insomnia-related DEGs. 287 shared targets were identified between Ganmai Dazao Decoction and insomnia. The traditional Chinese medicine-active ingredients-shared targets-disease network showed that quercetin， catechins and kaempferol were the key components of Ganmai Dazao Decoction in treating insomnia. These three components alleviate insomnia by acting on ten core targets， including nuclear factor kappa B inhibitor alpha （NFKBIA）， fibronectin 1 （FN1）， interleukin-6 （IL6）， protein c-Fos （FOS）， histone acetyltransferase p300 （EP300）， histone deacetylase 1 （HDAC1）， transcription factor Jun （JUN）， heat shock protein HSP 90-alpha 1 （HSP90AA1）， glyceraldehyde 3-phosphate dehydrogenase （GAPDH）， and interleukin-1 beta （IL1β）. GO and KEGG enrichment analyses indicated that Ganmai Dazao Decoction may alleviate insomnia through the IL17 signaling pathways， lipid and atherosclerosis signaling pathways， and other mechanisms. The results of molecular docking demonstrated strong binding affinity between the 3 key components and the 10 core targets of Ganmai Dazao Decoction. Molecular dynamics simulations further confirmed the stability of the quercetin-GAPDH， catechin-HDAC1 and kaempferol-EP300 complexes.Conclusion The key components of Ganmai Dazao Decoction， namely quercetin， catechin， and kaempferol， exert therapeutic effects on insomnia by targeting 10 core proteins and modulating multiple pathways， including the IL17 signaling pathway， lipids and atherosclerotic-related pathways. ［Funded by Chengdu Medical College Level Scientific Research Project （number， CYZYB23-01）］