基于网络药理学及分子对接技术研究灵芝治疗抑郁症的活性成分及其作用机制

Study on the Active Components and Mechanism of Ganoderma in the Treatment of Depression Based on Network Pharmacology and Molecular Docking Technology

摘要: 目的：基于网络药理学方法与分子对接技术对灵芝发挥抗抑郁作用的活性成分及其作用机制进行研究。方法：通过TCMSP、BATMAN-TCM数据库检索得到灵芝活性成分及相关靶点，通过DisGeNET、GeneCards、OMIM、DrugBank、TTD数据库以获取抑郁症的相关靶点。利用Venny2.1.0在线软件取抑郁症与灵芝靶点的交集，将共有靶点输入STRING在线分析平台得到灵芝与抑郁症蛋白质相互作用网络，再导入Cytoscape3.8.0软件构建灵芝活性成分–灵芝与抑郁症共有靶点关系网络，并进行模块化分析。利用DAVID6.8软件进行GO生物进程与KEGG通路富集分析，并使用R Studio软件画出通路的气泡图。最后通过分子对接验证网络分析结果，在已有数据中筛选灵芝抗抑郁作用的关键成分及靶点。结果：通过构建灵芝活性成分–灵芝与抑郁症共有靶点网络分析发现：灵芝与抑郁症相关的活性成分有40种，对应的基因靶点有210个，为潜在的抗抑郁症靶点。根据PPI网络可知：灵芝发挥抗抑郁作用主要与INS、TNF、FOS、PPARG、IL1B、ESR1、JUN、SLC6A4、LEP、CASP3 10个靶点有关。麦角胺、灵芝醇D、富马酸、灵芝酸甲酯F、5,6-麦角淄醇、赤灵芝酸D2甲酯、灵芝酸C、甲基灵芝F、麦角固醇、灵芝萜酮三醇10种为主要的活性成分。通过GO功能富集得到细胞组成73个，生物进程692个，分子功能131个，KEGG通路分析获得76条。GO与KEGG通路主要富集于信号传导、质膜的组成部分、药物反应、细胞增殖的正调控、c-AMP信号通路、神经活性配体–受体相互作用，提示灵芝可能通过以上多种通路达到治疗抑郁症的效果。分子对接结果提示麦角胺、灵芝醇D作为灵芝潜在的活性成分，与靶点FOS均有良好的结合力。结论：根据本次研究结果推测灵芝治疗抑郁症主要与其活性成分麦角胺、灵芝醇D有关。并且，首次将灵芝发挥抗抑郁作用与麦角胺、灵芝醇D和FOS的良好结合能力联系起来，预测麦角胺可作用于FOS以调节单胺类神经递质假说5-HT而发挥作用，灵芝醇D可作用于FOS以调控胆碱能肾上腺素能功能平衡失调而发挥作用。

Abstract: Objective: To study the antidepressant active components of Ganoderma and their mechanism based on network pharmacology and molecular docking technology. Methods: the active ingredients and related targets of Ganoderma were retrieved from TCMSP and BATMAN-TCM databases, and the related targets of depression were obtained from DisGeNET, GeneCards, OMIM, DrugBank and TTD databases. Take the intersection of depression and Ganoderma targets by using Venny2.1.0 online software, input the common targets into STRING online analysis platform to obtain the protein interaction network between Ganoderma and depression, and then import Cytoscape3.8.0 software to construct the Ganoderma and depression common target network, and conduct modular analysis. DAVID6.8 software is used for GO and KEGG pathway enrichment analysis, and R Studio software is used to draw the bubble diagram of the pathway. Finally, the results of network analysis were verified by molecular docking, and the key components and targets of Ganoderma anti depression were screened from the existing data. Results: through the construction of Ganoderma active components-Ganoderma and depression common target network analysis, it was found that there were 40 Ganoderma and depression related active components, corresponding to 210 gene targets, which were potential anti depression targets. According to PPI network, the antidepressant effect of Ganoderma is mainly related to 10 targets: INS, TNF, FOS, PPARG, IL1B, ESR1, JUN, SLC6A4, LEP and CASP3. Ergotamine, Fumaric Acid, Ganoderiol D, Methyl Ganoderate F, Methyl Lucidenate D2, Methyl Lucidenate F, Ganoderic Acid C, Ganodermanontriol, 5,6-Dihydroergosterol, Ergosterol are the main active ingredients. 73 cell components, 692 biological processes and 131 molecular functions were obtained by GO functional enrichment, and 76 KEGG pathways were obtained by KEGG pathway analysis. GO and KEGG pathways are mainly concentrated in signal transduction, components of plasma membrane, drug response, positive regulation of cell proliferation, c-AMP signal pathway, and neuroactive ligand receptor interaction, suggesting that Ganoderma may achieve the effect of treating depression through the above multiple pathways. The results of molecular docking showed that Ergotamine, Ganoderiol D, as potential active components of Ganoderma, had good binding ability with the target FOS. Conclusion: according to the results of this study, it is speculated that the treatment of depression by Ganoderma is mainly related to Ergotamine and Ganoderiol D. Moreover, for the first time, the antidepressant effect of Ganoderma was linked with the good binding ability of Ergotamine, Ganoderiol D and FOS. It was predicted that Ergotamine could act on FOS to regulate the monoamine neurotransmitter hypothesis 5-HT, and Ganoderiol D could act on FOS to regulate the imbalance of cholinergic and adrenergic functions.