基于网络药理学和分子对接技术分析除烦安神汤治疗失眠合并焦虑抑郁的作用机制

Analysis of Mechanism of Action of Chufan Anshen Decoction in the Treatment of Insomnia with Anxiety and Depression Based on Network Pharmacology and Molecular Docking Technology

摘要:

目的：通过网络药理学与分子对接技术分析张永华教授经验方除烦安神汤治疗失眠合并焦虑抑郁的潜在作用机制。方法：运用中药系统药理学数据库与分析平台（TCMSP）、中药分子机制生物信息学分析工具（BATMAN-TCM） 筛选除烦安神汤的活性成分；通过UniProt数据库将活性成分对应的蛋白靶点标准化为基因靶点。在GeneCards、OMIM数据库中检索失眠、焦虑、抑郁的相关靶点，经去重、筛选后构建疾病靶点集；利用R语言获取药物靶点与疾病靶点的交集靶点，借助Cytoscape 3.8.2软件构建“药物-成分-靶点”网络，通过拓扑分析筛选主要活性成分；通过STRING数据库构建蛋白质互作（PPI） 网络，结合MCODE插件筛选核心靶点；采用R语言对交集靶点进行基因本体（GO） 功能分析与京都基因与基因组百科全书（KEGG） 通路富集分析；并对核心活性成分与核心靶点进行分子对接验证。结果：共筛选出除烦安神汤活性成分516个，作用靶点1 484个；获得抑郁、焦虑、失眠靶点分别为5 443个、4 227个、479个，药物与疾病交集靶点143个。筛选出主要活性成分为槲皮素、薯蓣皂苷、小檗碱、大黄素、维生素E。核心靶点为肿瘤坏死因子（TNF）、白细胞介素-6（IL-6）、白细胞介素-1β （IL-1β）、原癌基因酪氨酸蛋白激酶（SRC）、表皮生长因子受体（EGFR）、γ-干扰素（IFNG） 和白细胞介素-8（CXCL8）。GO分析结果显示，潜在靶点生物学过程主要富集于对外源性刺激/脂多糖的反应、氧化应激调控、丝裂原活化蛋白激酶（MAPK） 级联调控；细胞组分主要涉及囊泡腔、膜筏、突触膜；分子功能集中于神经递质受体活性、配体门控离子通道活性、细胞因子受体结合。KEGG分析结果显示，潜在靶点主要富集于神经活性配体-受体相互作用通路、磷脂酰肌醇3激酶-蛋白激酶B（PI3K-AKT）信号通路、糖尿病并发症中的晚期糖基化终末产物及其受体（AGE-RAGE） 信号通路、TNF信号通路。主要活性成分薯蓣皂苷与核心靶点SRC、EGFR、TNF、IL-6、IL-1β的分子对接结合能均＜-5 kcal/mol，其中与SRC结合能最低为-8.9 kcal/mol。结论：除烦安神汤治疗失眠合并焦虑抑郁可能通过槲皮素、薯蓣皂苷等活性成分，作用于TNF、IL-6、SRC等核心靶点，调控神经活性配体-受体相互作用、PI3K-AKT、AGE-RAGE等关键通路，协同发挥抑制神经炎症、调节神经信号传导、改善氧化应激的作用，从而实现对失眠合并焦虑抑郁情绪与睡眠症状的双重改善。

Abstract:

Abstract：Objective：To analyze the potential mechanism of action of Professor ZHANG Yonghua's experienced prescription， Chufan Anshen Decoction， in the treatment of insomnia with anxiety and depression using network pharmacology and molecular docking technology. Methods： Active components of Chufan Anshen Decoction were screened using the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform （TCMSP） and the Bioinformatics Analysis Tool for Molecular mechANism of Traditional Chinese Medicine （BATMAN-TCM）. The protein targets corresponding to the active components were standardized into gene targets via the Universal Protein Resource （UniProt） database. Targets related to insomnia， anxiety， and depression were retrieved from the Human Gene Database （GeneCards） and Online Mendelian Inheritance in Man （OMIM） databases. After deduplication and screening， a disease target set was constructed. Intersection targets between drug targets and disease targets were obtained using R language. The "drug-component-target" network was constructed using Cytoscape 3.8.2 software，and core active components were screened through topological analysis. A protein-protein interaction （PPI） network was constructed using the Search Tool for the Retrieval of Interacting Genes/Proteins （STRING） database，and core targets were screened using the MCODE plug-in. Gene Ontology （GO） functional analysis and Kyoto Encyclopedia of Genes and Genomes （KEGG） pathway enrichment analysis of the intersection targets were performed using R language. Molecular docking validation between core active components and core targets was then conducted. Results：A total of 516 active components and 1 484 corresponding targets of Chufan Anshen Decoction were screened. A total of 5 443 targets for depression，4 227 targets for anxiety and 479 targets for insomnia were obtained，yielding 143 intersection targets between drugs and diseases. The main active components identified were quercetin，dioscin，berberine，emodin，and vitamin E. The core targets were tumor necrosis factor （TNF）， interleukin-6 （IL-6）， interleukin-1β （IL-1β）， proto-oncogene tyrosineprotein kinase （SRC），epidermal growth factor receptor （EGFR），interferon gamma （IFNG）， and interleukin-8 （CXCL8）. GO analysis showed that the biological processes of potential targets were mainly enriched in response to xenobiotic stimulus/lipopolysaccharide， regulation of oxidative stress， and regulation of the mitogenactivated protein kinase （MAPK） cascade. Cellular components mainly involved vesicle lumen，membrane raft， and synaptic membrane. Molecular functions focused on neurotransmitter receptor activity， ligandgated ion channel activity，and cytokine receptor binding. KEGG analysis revealed that the potential targets were mainly enriched in the neuroactive ligand-receptor interaction pathway， the phosphatidylinositol 3-kinase protein kinase B （PI3K-AKT） signaling pathway， the advanced glycation end products receptor for advanced glycation end products （AGER-AGE） signaling pathway in diabetic complications， and the TNF signaling pathway as key pathways. The main active component，dioscin，exhibited molecular docking binding energies of less than -5 kcal/mol with the core targets SRC， EGFR，TNF，IL-6，and IL-1β，the lowest being -8.9 kcal/mol with SRC. Conclusion：Chufan Anshen Decoction may exert its therapeutic effects on insomnia with anxiety and depression through active components such as quercetin and dioscin，acting on core targets including TNF，IL-6，and SRC，and regulating key pathways such as neuroactive ligand receptor interaction， PI3K-AKT， and AGE-RAGE. It synergistically inhibits neuroinflammation， modulates neural signal transmission， and ameliorates oxidative stress， thereby achieving dual improvement of emotional symptoms and sleep disturbances in insomnia with anxiety and depression.