摘要
基于网络药理学及分子对接方法筛选马氏珍珠贝软体来源的具有潜在降糖活性的肽类成分。采用纳升液相串联质谱鉴定马氏珍珠贝软体中的主要蛋白质类成分;通过Peptide Cutter在线工具模拟酶切获得马氏珍珠贝软体来源的肽类成分;进一步借助SYBYL-X 2.0软件进行分子对接筛选潜在活性肽段;通过Swiss Target数据库收集活性肽段潜在靶点,采用GeneCards数据库和OMIM-GENE-MAP数据库收集2型糖尿病相关的疾病靶点;借助Cytoscape软件和STRING平台构建“肽段-成分-靶点”网络和蛋白质相互作用网络,进行可视化分析,筛选酶解肽的核心成分和关键靶点;利用Omicshare云平台对共同靶点进行GO(Gene Ontology)和KEGG(Kyoto Encyclopedia of Genes and Genomes)富集分析;体外二肽基肽酶-IV(dipeptidylpeptidase-IV,DPP-IV)抑制率实验验证马氏珍珠贝软体肽段的活性。结果表明,肌动蛋白、肌球蛋白和微管蛋白为马氏珍珠贝软体主要蛋白质类成分,从模拟酶切肽段中筛选获得潜在降糖活性肽段28条及其对应基因靶点377个;确定2型糖尿病疾病靶点1705个;可视化分析筛选出25个关键靶点;富集分析显示,马氏珍珠贝软体酶解肽主要参与细胞增殖与凋亡、蛋白质代谢、信号转导、炎症反应等过程。活性验证结果表明,19条肽段具有体外DPP-IV抑制活性,其中26号活性肽段体外DPP-IV抑制活性最佳(IC_(50)=395μmol/L)。网络药理学和分子对接方法可用于筛选马氏珍珠贝软体中活性肽类成分,据此筛选的活性肽类成分可通过调控炎症、物质代谢、细胞凋亡等途径发挥降糖作用。
Based on network pharmacology and molecular docking techniques,peptides with potential anti-diabetic were screened from Pinctada martensii flesh.Nano-LC Q Exactive Orbitrap mass spectrometry(nano LC-MS/MS)was used to identify the major proteins in the P.martensii flesh.The predicted peptides were obtained under simulated enzyme digestion by using Peptide Cutter tool.Moreover,the potential active peptides were screened by molecular docking with SYBYL-X 2.0 software.The targets of potential active peptide were collected using Swiss Target database.Type 2 diabetes-related disease targets were collected based on the GeneCards database and OMIM-GENE-MAP database.Furthermore,cytoscape software and STRING platform were used to construct the peptide-component-target’network and protein interaction network,and visualized analysis was performed to screen the core components and key targets of enzymatic peptides.Gene Ontology(GO)enrichment and Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment analysis of common targets were carried out through Omicshare cloud platform.The dipeptidylpeptidase-IV(DPP-IV)inhibitory activity of peptides from P.martensii flesh was verified.The results showed that actin,myosin and tubulin were identified as the main components in the P.martensii flesh.Twenty-eight potential anti-diabetic peptides and their 377 gene targets were obtained.There were 1705 disease targets for type 2 diabetes were identified and 25 key targets were screened out by visual analysis.The enrichment analysis showed that the enzymatic hydrolysis peptides were mainly involved in cell proliferation and apoptosis,protein metabolism,signal transduction and inflammatory reaction.Among those potential anti-diabetic peptides,19 peptides showed DPP-IV inhibitory activity in vitro,peptide 26 showed the best DPP-IV inhibitory activity(IC_(50),395μmol/L).Active peptides could be screened by network pharmacology and molecular docking could be applied to screen active peptides from Pinctada martensii flesh.Based on this method,screened potential anti-diabetic peptides could exert their activity through pathways including regulating inflammation,substance metabolism and cell apoptosis.
作者
李佳芸
王欣之
韦源青
林彬燕
刘睿
吴皓
LI Jiayun;WANG Xinzhi;WEI Yuanqing;LIN Binyan;LIU Rui;WU Hao(Jiangsu Key Laboratory of Research and Development in Marine Bio-resource Pharmaceutics,Nanjing 210023,China;College of Pharmacology,Nanjing University of Chinese Medicine,Nanjing 210023,China;Jiangsu Collaborative Innovation Center of Chinese Medicinal Resources Industrialization,Nanjing 210023,China)
出处
《食品与发酵工业》
CAS
CSCD
北大核心
2022年第15期176-184,共9页
Food and Fermentation Industries
基金
药用南珠标准化养殖与综合利用产业链协同创新项目(BHsfs002)
国家自然科学基金面上项目(81973450)
江苏省海洋科技创新专项(HY2017-7)
江苏省高等学校自然科学研究重大项目(17KJA360005)
中华中医药学会“青年人才托举工程”项目(QNRC2-C14)
江苏省高校“青蓝工程”中青年学术带头人项目
南京中医药大学中药学一流学科开放课题项目(2020YLXK009)。
关键词
马氏珍珠贝软体酶解肽
2型糖尿病
网络药理学
分子对接
Pinctada martensii flesh enzyme peptides
type 2 diabetes
network pharmacology
molecular docking
