基于酶抑制实验与网络药理学探讨核桃楸皮总黄酮治疗2型糖尿病的作用机制  

Study on the Action Mechanism of Total Flavonoids of Catalpa Juglans Peel in the Treatment of Type 2 Diabetes Mellitus Based on Enzyme Inhibition Experiment and Network Pharmacology

作　　者：贾凯杰 韩翔宇 于慧[1] 赵盼[1] JIA Kaijie;HAN Xiangyu;YU Hui;ZHAO Pan(Shandong University of Traditional Chinese Medicine,Jinan,Shandong,China,250355)

机构地区：[1]山东中医药大学,山东济南250355

出　　处：《河南中医》2023年第10期1518-1525,共8页Henan Traditional Chinese Medicine

基　　金：国家自然科学基金项目(81903780);山东中医药大学大学生创新创业训练计划项目(2022042)。

摘　　要：目的:通过酶抑制实验考察核桃楸皮总黄酮的降糖作用,并利用网络药理学与分子对接的方法研究核桃楸皮总黄酮治疗2型糖尿病的潜在作用机制。方法:提取纯化核桃楸皮总黄酮部位后采用DNS法测定核桃楸皮总黄酮对α-淀粉酶的抑制作用。将文献报道的核桃楸皮的27种黄酮类成分输入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)进行筛选。将筛选后的活性成分导入Swiss Target Prediction数据库进行靶点预测。将核桃楸皮黄酮类活性成分与相关靶点导入Cytoscape 3.9.1软件构建“化合物-靶点”网络,并筛选主要活性成分。在Genecards、TTD、DisGeNET数据库中搜集2型糖尿病相关的靶点,利用在线工具Venny 2.1得到活性成分靶点和疾病靶点的交集靶点,并绘制Venn图。在STRING数据库中输入交集靶点得到网络关系,利用Cytoscape 3.9.1软件构建PPI网络并进行拓扑分析。通过Metascape分析平台对交集靶点进行基因本体(gene ontology, GO)富集分析和京都基因与基因组百科全书(kyoto encyclopedia of genes and genomes, KEGG)信号通路富集分析。利用AutoDock Vina 1.2.0对关键成分和核心靶点进行分子对接。结果:核桃楸皮总黄酮粉末纯度为93.85%,核桃楸皮总黄酮对α-淀粉酶具有明显的抑制作用(P<0.05),IC_(50)值为0.776 g·L^(-1)。从TCMSP数据库筛选得到9种活性成分,获得223个活性成分相关靶点。“化合物-靶点”网络分析得到山柰酚、木犀草素、杨梅素、柚皮素、槲皮素可能是发挥作用的主要活性成分。在Genecards、TTD、DisGeNET数据库获得疾病靶点1 360个。利用在线工具Venny 2.1进行取交集处理,得到交集靶点99个。PPI网络分析得到蛋白激酶B1(protein kinase B1,AKT1)、肿瘤坏死因子(tumor necrosis factor, TNF)、血管内皮生长因子A(vascular endothelial growth factor A,VEGFA)、表皮生长Objective:To investigate the hypoglycemic effect of total flavonoids of Catalpa juglans peel by enzyme inhibition experiment,and study its potential mechanism in the treatment of type 2 diabetes mellitus by means of network pharmacology and molecular doc-king.Methods:After extraction and purification of total flavonoids from Catalpa juglans peel,the inhibitory effect of total flavonoids onα-amylase was determined by the DNS method.A total of 27 flavonoid components reported in the literature were input into the traditional Chinese medicine systems pharmacology database and analysis platform(TCMSP)for screening.The screened active components were imported into the Swiss Target Prediction database for target prediction.Flavonoids and related targets were imported into Cytoscape 3.9.1 software to construct a"compound-target"network,and the main active components were screened.Targets related to type 2 dia-betes were collected from Genecards,TTD and DisGeNET databases,and the intersection targets of active component targets and disease targets were obtained by using the online tool Venny 2.1,and the Venn map was drawn.The intersection targets were entered into the STRING database to obtain the network relationship.The PPI network was constructed by Cytoscape 3.9.1 software and topological a-nalysis was carried out.Gene ontology(GO)enrichment analysis and Kyoto encyclopedia of genes and genomes(KEGG)signaling pathway enrichment analysis were conducted for the intersection targets through the Metascape analysis platform.Auto Dock Vina 1.2.0 was used for molecular docking of key components and core targets.Results:The purity of total flavonoid was 93.85%,and total fla-vonoid had obvious inhibitory effect onα-amylase(P<0.05),and ICso value was 0.776 g·L^(-1).A total of 9 active components were screened from TCMSP database,and 223 active component-related targets were obtained.The"compound-target"network analysis showed that kaempferol,luteolin,myricetin,naringin and quercetin might be the main active components.A tota

关 键 词：2型糖尿病 核桃楸皮总黄酮 酶抑制实验 网络药理学 分子对接 作用机制 

分 类 号：R285.5[医药卫生—中药学]