益肾化浊方治疗阿尔茨海默病的网络药理学分析及实验验证

Mechanism of Yishen Huazhuo Decoction in Treatment of Alzheimer ""s disease based on network pharmacology and molecular docking technology

关键词:  益肾化浊方  阿尔茨海默病  网络药理学  分子对接  实验验证

Key words:  Yishen Huazhuo Decoction  Alzheimer

摘要:

目的 基于网络药理学方法及分子对接技术探究益肾化浊方治疗阿尔茨海默病（Alzheimer’s disease, AD）的机制并予以实验验证。方法 通过中药系统药理学数据库与分析平台（TCMSP）与文献检索复方组分及靶标，通过人类基因综合分析（GeneCards）数据库检索疾病靶标，绘制韦恩图，提取复方组分与疾病的交集靶点；利用STRING数据库构建蛋白互作网络（PPI）；运用Cytoscape3.7.2，构建“中药复方－疾病－交集靶标”网络；运用DAVID数据库进行基因本体论（GO）功能分析和京都基因与基因组百科全书（KEGG）通路富集分析。借助AutoDock Tools分子模拟软件构建分子对接。以APP/PS1转基因小鼠作为研究对象，HE染色法检测小鼠海马组织病理学改变；Western Blot与PCR检测小鼠海马组织中PI3K/AKT/GSK-3β信号通路关键因子的表达。结果 经网络药理学筛选获得核心靶标有AKT1、TNF、IL6等，信号通路主要包括PI3K/AKT等；分子对接表明，山柰酚、槲皮素、β-谷甾醇等与靶标具有较高亲和力；动物实验验证结果显示，益肾化浊方可减轻小鼠海马组织病理学改变；上调PI3K/AKT/GSK-3β信号通路关键蛋白的表达。结果 益肾化浊方能够改善AD小鼠的神经损伤，其机制与激活中枢PI3K/AKT/GSK-3β信号通路相关，本研究为临床治疗AD提供理论基础。

Abstract:

OBJECTIVE The study aims to investigate the mechanism of Yishen Huazhuo Decoction (YHD) in treating Alzheimer’s disease (AD) through network pharmacology and molecular docking techniques. METHODS The pharmacology database and analysis platform of traditional Chinese medicine system (TCMSP) and literature search were used to retrieve the components and targets of the compound prescription, and the GeneCards database was used to retrieve the disease targets. Draw Wayne diagram to extract the intersection targets of compound components and diseases; Using the STRING database to construct protein interaction network (PPI); Cytoscape3.7.2 was used to construct the network of “TCM compound - disease - intersection target”. The DAVID database was used for enrichment analysis of Gene Ontology (GO) function analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis. Build molecular docking using AutoDock Tools molecular simulation software. In this study, APP/PS1 transgenic mice were used as research objects, and the histopathological changes of hippocampus were detected by HE staining. Western Blot and PCR were used to detect the expression of key factors of PI3K/AKT/GSK-3β signaling pathway in the hippocampus of mice. RESULTS The core targets obtained through network pharmacology screening include AKT1, TNF, IL6 and other core targets, and the signaling pathways mainly included TNF, PI3K/AKT and so on. Molecular docking results showed that kaempferol, quercetin and β-sitosterol had high affinity to the targets. The results of animal experiments showed that YHD could alleviate the histopathological changes in the hippocampus of mice. Up-regulated expression of PI3K/AKT/GSK-3β signaling pathway. CONCLUSION The Yishen Huazhuo Decoction could improve nerve damage in AD mice, and its mechanism was related to the activation of the PI3K/AKT/GSK-3β signaling pathway, thereby offering a theoretical foundation for its clinical application.