基于网络药理学与秀丽隐杆线虫模型探讨黄芪总黄酮治疗阿尔茨海默病作用机制

Exploration of the Mechanism of Total Flavonoids of Astragalus in Treatment of Alzheimer's Disease Based on Network Pharmacology,Molecular Docking and Caenorhabditis Elegans Model

目的:采用网络药理学与分子对接技术探讨黄芪总黄酮(TFA)治疗阿尔茨海默病(AD)的潜在作用机制,并利用转基因秀丽隐杆线虫模型进行初步药效验证。方法:利用TCMSP和SwissTargetPrediction数据库获得TFA的有效成分和潜在靶点,通过OMIM和GeneCards数据库获得AD的潜在靶点;采用Venny 2.1.0软件对TFA与AD进行相互影响的综合评估,同时以STRING 12.0数据库建立蛋白质互作网络;采用DAVID 6.8数据库进行GO和KEGG的重要目标的聚类分析;利用Cytoscape 3.7.1创建TFA治疗AD的“组成部分-目标-路径”网络结构;并使用AutoDock Tools工具进行分子链接。利用CL4176线虫模型,探究TFA对线虫寿命、运动能力、抑制瘫痪、抗热应激、抗氧化应激能力、抑制β-淀粉样蛋白聚集的作用。结果:TFA对抗AD的有效成分共9个,交集靶点共106个;GO功能富集分析得到732个条目;KEGG通路富集得到156条信号通路;“成分-靶点-通路”网络关系图及分子对接结果显示,AKT1、TNF、IL-6等靶点可能为TFA治疗AD的关键靶点;线虫实验结果显示,TFA具有提高线虫寿命、运动能力、抗热应激、抗氧化应激能力,并抑制瘫痪与β-淀粉样蛋白聚集的作用。结论:TFA中华良姜素、异鼠李素等活性成分通过AKT1、TNF、IL-6等核心靶点与PI3K-Akt、MAPK、AGE-RAGE、IL-17等信号通路参与治疗AD,为其后续理论与临床研究提供参考。

Objective:Exploring the potential mechanism of total flavonoids of Astragalus for Alzheimer's disease,network pharmacology and molecular docking were adopted.To verify the preliminary efficacy through the transgenic Caenorhabditis elegans model.Methods:The active components of total flavonoids of Astragalus and their potential targets were obtained from the database analysis platform of traditional Chinese medicine system pharmacology and SwissTargetPrediction database.OMIM and GeneCards database were used to obtain disease targets of Alzheimer's disease.Venny 2.1.0 was used to obtain the intersection of drug targets and potential disease targets,and the protein-protein interaction network was constructed with STRING 12.0 database.Gene Ontology(GO)enrichment analysis and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analysis were performed on key targets through DAVID 6.8 database.The“constituent-target-pathway”network of total flavonoids of Astragalus in the treatment of Alzheimer's disease was constructed by Cytoscape 3.7.1.AutoDock Tools software was used to perform molecular docking between key components and core targets.Through the CL4176 Caenorhabditis elegans model,the effects of total flavonoids of Astragalus on the lifespan,exercise capacity,inhibition of paralysis,heat stress resistance,oxidative stress resistance,and inhibition ofβ-amyloid aggregation were investigated.Results:There were 9 effective components and 106 intersection targets of total flavonoids of Astragalus against Alzheimer's disease.GO enrichment analysis yielded 732 results.KEGG pathway enrichment obtained 156 signal pathways.The results of“constituent-target-pathway”network diagram and molecular docking showed that AKT1,TNF,IL6 and other targets may be the key targets of total flavonoids of Astragalus in the treatment of Alzheimer's disease.The results of nematode experiment showed that total flavonoids of Astragalus could improve the lifespan,exercise ability,anti-heat stress,anti-oxidative stress ability of Caenorhabditis elegans,and inhibit paralysis and Amyloid-βaggregation.Conclusion:The study showed that the active ingredients of TFA Chinese galangal and isorhamnetin were involved in the treatment of AD through the core targets such as AKT1,TNF and IL6 and signaling pathways such as PI3K-Akt,MAPK,AGE-RAGE and IL-17,which provided a reference for its subsequent theoretical and clinical studies.