基于网络药理学方法探讨黄芪治疗骨骼肌减少症的作用机制

Network Pharmacology-based Study on the Mechanism of Huangqi in the Treatment of Sarcopenia

目的:使用网络药理学方法分析黄芪治疗骨骼肌减少症的作用机制。方法:使用中药系统药理学数据库与分析平台(traditional Chinese medicine systems pharmacology database and analysis platform,TCMSP)筛选黄芪的活性成分并预测其作用靶点;筛选GEO与人类基因数据库(the human gene database,GeneCards)中有关骨骼肌减少症的相关基因并进行合并;使用STRING在线数据库将疾病与药物交集靶点进行蛋白相互作用分析,并使用Cytoscape 3.7.2软件对蛋白-蛋白互作网络(protein-protein interactions,PPI)进行拓扑学分析,确定核心靶点;使用Cytoscape软件构建“药物-活性成分-靶点-疾病”网络,使用AutoDockTools等软件对药物关键分子与疾病关键蛋白进行分子对接验证;最后利用DAVID数据库对潜在靶点进行基因本体论(gene ontology,GO)分析,探究黄芪治疗骨骼肌减少症的生物功能,通过京都基因与基因组百科全书(kyoto encyclopedia of genes and genomes,KEGG)通路富集分析探究黄芪治疗骨骼肌减少症的主要信号通路。结果:共得到黄芪有效活性成分作用靶点155个,骨骼肌减少症相关基因靶点1057个,药物-疾病交集靶基因36个,主要包括Akt1、p53、VEGFA、TNF、ESR1等“;药物-活性成分-靶点-疾病”调控网络显示槲皮素、华良姜素、联苯双酯、7-O-methylisomucronulatol、异鼠李素等药物活性成分在治疗骨骼肌减少症中发挥作用;分子对接验证实现了5种主要活性成分与5种关键靶点蛋白的对接,结果发现其均具有较强的结合活性;GO分析结果显示,黄芪治疗骨骼肌减少症主要与调控炎症反应及凋亡过程的正调控、细胞对缺氧的反应、细胞衰老等生物学过程有关,并影响蛋白质结合、转运活性、过氧化物酶体增殖物激活受体结合、肌动蛋白结合、生长因子活性、蛋白磷酸酶抑制剂活性等分子功能;KEGG分析显示靶点基因主要富集在癌症、肿瘤坏死因子、低氧诱导因子-1、细胞凋亡等通路。结论:黄芪可能通过同时调控多个靶点基因及相关信号通路,发挥诱导细胞衰老和凋亡、改善局部微循环、降低细胞外基质降解速度、减轻慢性炎症及氧化应激损伤等作用,以延缓骨骼肌萎缩。

Objective:To analyze the mechanism of Huangqi(Astragali radix)in the treatment of sarcopenia using network pharmacology.Methods:TCMSP was used to search active ingredients of Huangqi,and predict its targets;the genes related to sarcopenia were screened from GEO and GeneCards databases,and combined.STRING online database was used to perform protein-protein interaction of the intersected targets between disease and medicine,Cytoscape 3.7.2 software was applied to conduct topological analysis of PPI network,thereby confirming the core targets;Cytoscape software was utilized to construct regulation network of"medicine-active ingredients-targets-disease",and determine the importance of each network node;AutoDockTools was adopted to perform the verification of molecular docking of key molecules of medicine and key protein of disease;consequently,DAVID database was applied to carry out GO analysis of the potential targets,to survey the biological function of Huangqi in the treatment of sarcopenia,and KEGG pathway enrichment analysis was used to find out the main signal pathways of Huangqi in the treatment of sarcopenia.Results:Finally the study has identified 155 targets of active ingredients of Huangqi,1057 genetic targets related to sarcopenia and 36 intersected genes between drug and diseases,including Akt1,p53,VEGFA,TNF,ESR1 and others;"drug-active ingredients-targets-disease"regulation network displayed that active ingredients of drug such as quercetin,jaranol,bifendate,7-O-methylisomucronulatol and isorhamnetin played the roles in the treatment of sarcopenia;molecular docking validation achieves docking of the five main active ingredients to five key target proteins,which showed that they had strong binding activities;GO analysis results showed that Huangqi in the treatment of sarcopenia mainly participated in biological processes,including the regulation of inflammatory reaction,positive regulation of apoptotic processes,cellular response to hypoxia,and cellular aging,influencing molecular functions of protein binding,transporter activity,peroxisome proliferator-activated receptor binding,actin binding,growth factor activity and protein phosphatase inhibitor activity;KEGG analysis indicated that target genes were mainly enriched in cancer pathway,TNF signaling pathway,hypoxia-inducible factor-1 signaling pathway,and cellular apoptosis.Conclusion:Huangqi could play the roles in regulating cellular aging and apoptosis,improving local microcirculation,reducing the rate of extracellular matrix degradation,relieving chronic inflammation and oxidative stress damage possibly through regulating targets genes and the related signaling pathway via many active ingredients,so as to delay skeletal muscle atrophy.