基于网络药理学与分子对接探究苦虎益母口服液治疗鸡输卵管炎的作用机制

Exploring the Mechanism of Action of Kuhu Yimu Oral Liquid in the Treatment of Chicken Salpingitis Based on Network Pharmacology and Molecular Docking

关键词:  网络药理学  分子对接  苦虎益母口服液  鸡输卵管炎

Key words:  network pharmacology  molecular docking  Kuhu Yimu oral liquid  chicken salpingitis

摘要:

为了探讨苦虎益母口服液治疗鸡输卵管炎的作用机制，试验通过网络药理学和分子对接的方法进行研究。首先筛选TCMSP数据库中苦参、虎杖、黄芪、菟丝子、益母草五味药材的成分和靶点，利用Uniprot数据库对靶标进行规范化处理，利用Cytoscape 3.8.2建立药物-有效活性成分-靶点网络图。由GeneCards、OMIM、CTD数据库收集鸡输卵管炎的靶点。利用微生信在线平台制作鸡输卵管炎和苦虎益母口服液的韦恩图，找到共同靶点。由STRING数据库构建蛋白质-蛋白质互作(PPI)网络互作图，并通过Cytoscape 3.8.2可视化。再使用DAVID在线工具对重叠靶标进行基因本体（GO）和京都基因和基因组百科全书（KEGG）通路富集分析。使用AutoDock Tools软件对核心成分和靶点进行分子对接验证。预测苦虎益母口服液治疗鸡输卵管炎的活性成分有74种，对应靶点77个，1137个鸡输卵管炎靶点，35个共同靶点。PPI网络显示涉及的关键靶点是雌激素受体1 (ESR1)、细胞周期调节因子（CCND1）、血管内皮生长因子A（VEGFA）。GO功能富集分析筛选符合伪发现率<0.05的条件后得到相关条目8个。生物过程(biological process, BP)涉及到白细胞介素-18、巨噬细胞活化、免疫反应、细胞对异生刺激的反应、RNA聚合酶II启动子转录正调；细胞组分(cellular component, CC)涉及到细胞外间隙；分子功能(molecular function, MF)涉及到干扰素-γ受体结合、细胞因子。KEGG富集分析表明苦虎益母口服液发挥作用的通路主要有细胞衰老、细胞周期和Toll样受体信号通路。分子对接结果表明苦虎益母口服液主要活性有效活性成分槲皮素、山奈酚和木犀草素与关键靶点ESR1、CCND1和VEGFA的结核活性较好，结合能均小于0 kJ/mol。苦虎益母口服液有的多种有效成分可能作用于多个靶点，通过参与多种信号通路对鸡输卵管炎发挥作用。

Abstract:

In order to investigate the mechanism of action of Kuhu Yimu oral liquid in the treatment of oviductal inflammation in chickens, the experiment was investigated by network pharmacology and molecular docking. Firstly, we screened the components and targets of five herbs, namely, Bitter Ginseng, Tiger Balm, Astragalus, Cuscuta, and Motherwort, in TCMSP database, and normalized the targets by using Uniprot database, and established the network diagram of drug-active ingredient-target by using Cytoscape 3.8.2. Targets for chicken vasculitis were collected by GeneCards, OMIM, and CTD databases. A Wayne diagram of chicken vasculitis and Kuhu Yimu oral liquid was created using the Microbiology online platform to find common targets. Protein-protein interaction (PPI) network interaction maps were constructed from the STRING database and visualized by Cytoscape 3.8.2. Overlapping targets were then analyzed for Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment using the DAVID online tool. Molecular docking validation of core components and targets using AutoDock Tools software. There are 74 active ingredients predicted to be used in the treatment of chicken vasculitis by Kuhu Yimu oral liquid, corresponding to 77 targets, 1137 chicken vasculitis targets and 35 common targets. The key targets shown to be involved by the PPI network are estrogen receptor 1 (ESR1), cell cycle regulator (CCND1), and vascular endothelial growth factor A (VEGFA). GO functional enrichment analysis screened for compliance with a pseudo-discovery rate of <0.05 yielded 8 relevant entries. Biological process (BP) involves interleukin-18, macrophage activation, immune response, cellular response to xenobiotic stimuli, and transcriptional up-regulation of RNA polymerase II promoter; cellular component (CC) involves extracellular space; and molecular function (MF) involves interferon-gamma receptor binding, and cytokines. KEGG enrichment analysis showed that the pathways in which Kuhu Yimu oral liquid acted were mainly cellular senescence, cell cycle and Toll-like receptor signaling pathways. The results of molecular docking showed that quercetin, kaempferol and lignocerotoxin, the main active ingredients of Kuhu Yimu oral liquid, had good nodulation activity with ESR1, CCND1 and VEGFA, and the binding energies of these ingredients were less than 0 kJ/mol. Kuhu Yimu oral liquid has many active ingredients that may act on multiple targets, and may be able to play a role in the oviductal inflammation of chickens by participating in multiple signaling pathways.