Study on the mechanism of Guipi decoction in treating tinnitus based on network pharmacology and molecular docking
基于网络药理学和分子对接探讨归脾汤治疗耳鸣作用机制研究

摘要

目的 运用网络药理学方法探究归脾汤治疗耳鸣作用机制。方法 在TCMSP数据平台上归纳整理归脾汤组成的有效成分，通过中国知网上检索建库至2023年的文献进行补充，并将得到的有效成分导入Uniprot平台，转换成对应的蛋白质靶点，选定种类智人，且删除还未验证靶点，最后得到归脾汤药物作用靶点基因的名称。使用GeneCards、OMIM、PharmGkb、TTD、DrugBank 5个平台上检索出耳鸣疾病相关基因靶点，以Score值≥中位数的为标准进行筛选，获得的基因靶点为耳鸣基因的相关靶点。利用Venny2.1工具构建“药物—疾病”韦恩图以确定二者交集靶点，交集靶点通过STRING平台绘制蛋白相互作用网络图，保存为TSV格式。之后导入并利用Cytoscape软件选出核心靶点，进行GO、KEGG富集分析，最后进行分子对接。结果 归脾汤有效成分共计228个，主要作用靶点246个，主要活性成分为山柰酚、细叶远志皂苷、槲皮素等，耳鸣病主要靶点1938个，归脾汤-耳鸣病交集靶点83个，归脾汤治疗耳鸣病主要作用于CTNNB1、TNF、ALB、EGFR、PPARγ、IL6、HIF-1α、VEGFA、CASP3、IL1B核心靶点，靶点多富集于HCMV信号通路、Apoptosis信号通路、TNF信号通路等。分子对接结果显示成分与核心靶点都能成功对接，且结构稳定。结论 归脾汤治疗耳鸣是通过保护耳蜗神经细胞、抗氧化激化反应、抗炎作用等作用，从而改善并减轻耳鸣患者症状。

Abstract

Objective To investigate the mechanism of action of Guipi Decoction in treating tinnitus using network pharmacology.
Methods The active ingredients of Guipi Decoction were summarized and organized on the TCMSP database platform. Literature from the China National Knowledge Infrastructure (CNKI) database up to 2023 was searched for these active ingredients. The resulting active ingredients were imported into the Uniprot platform and converted into corresponding protein targets. Homo sapiens species were selected, and unverified targets were deleted. Finally, the names of the target genes of Guipi Decoction were determined. Gene targets related to tinnitus were identified using GeneCards, OMIM, PharmGkb, TTD, and DrugBank. Gene targets were screened using a score ≥ median. The identified gene targets were identified as tinnitus-related targets. A drug-disease Venn diagram was constructed using the Venny 2.1 tool to identify drug-disease intersection targets. Protein interaction networks were then generated using the STRING platform and saved in TSV format. Core targets were then selected using Cytoscape software for GO and KEGG enrichment analysis, followed by molecular docking.
Results Guipi Decoction (GPD) contained 228 active ingredients and 246 primary targets, including kaempferol, polygala tenuifolia saponins, and quercetin. There were 1938 primary targets for tinnitus, including 83 targets that intersected between GPD and tinnitus. The GPD treatment of tinnitus primarily targeted CTNNB1, TNF, ALB, EGFR, PPARγ, IL6, HIF-1α, VEGFA, CASP3, and IL1B, with most targets enriched in the HCMV signaling pathway, apoptosis signaling pathway, and TNF signaling pathway. Molecular docking results demonstrated successful docking of the ingredients with the core targets, with stable structures.
Conclusion   Guipi Decoction treats tinnitus by protecting cochlear nerve cells, resisting oxidative stress, and exerting anti-inflammatory effects, thereby improving and alleviating tinnitus symptoms.