基于网络药理学和蛋白组学的地龙抗血栓作用机制研究  

Study on Mechanisms of Antithrombotic Ability of Dilong(Earthworms)Based on Network Pharmacology and Proteomics

作　　者：吴娅丽 马韫楠 李伟霞[1,3] 王晓艳[1,3] 张辉[1] 杨柳青 唐进法[1,3] 杜守颖[2] WU Yali;MA Yunnan;LI Weixia;WANG Xiaoyan;ZHANG Hui;YANG Liuqing;TANG Jinfa;DU Shouying(Henan Province Engineering Research Center of Clinical Application,Evaluation and Transformation of Traditional Chinese Medicine,Henan Provincial Key Laboratory for Clinical Pharmacy of Traditional Chinese Medicine,Henan Province Engineering Research Center of Safety Evaluation and Risk Management of TCM,The First Affiliated Hospital of Henan University of Chinese Medicine,Zhengzhou 450000,Henan,China;Beijing University of Chinese Medicine,Beijing 102488,China;Henan University of Chinese Medicine,Zhengzhou 450046,Henan,China)

机构地区：[1]河南中医药大学第一附属医院,河南省中药临床应用、评价与转化工程研究中心,河南省中药临床药学中医药重点实验室,河南省中药安全评价与风险防控工程研究中心,河南郑州450000 [2]北京中医药大学,北京102488 [3]河南中医药大学,河南郑州450046

出　　处：《中华中医药学刊》2023年第10期76-82,I0019,I0020,共9页Chinese Archives of Traditional Chinese Medicine

基　　金：国家自然科学基金项目(82104352);河南省中医药拔尖人才培养项目(豫中医科教[2018]35号);河南省高校科技创新团队项目(23IRTSTHN026)。

摘　　要：基于网络药理学和蛋白组学技术探究地龙抗血栓的物质基础及作用机制。首先通过网络药理学方法对地龙小分子活性成分进行研究,基于文献检索收集地龙化学成分101个,包括核苷酸、有机酸、氨基酸及寡肽、脂类及其他,构建成分数据库,预测成分作用靶标共798个;以课题组前期已验证的地龙抗血栓靶点(纤维蛋白原、纤维蛋白、纤溶酶原)为基础,通过BioGRID数据库挖掘与其具直接作用的靶点1046个,为潜在抗血栓靶点库,二者共有靶点81个;根据核心靶点蛋白互作网络及成分-靶点-通路网络分析,发现脂类、氨基酸和寡肽类成分为主要抗血栓成分,纤连蛋白、纤溶酶原、促分裂原活化蛋白激酶-14、基质金属蛋白酶-9、表皮生长因子受体、凝血酶原、纤维蛋白原等为地龙抗血栓核心靶点,通过调控黏着斑、白细胞跨内膜转移、补体与凝血、细胞周期及坏死性凋亡等发挥作用。继而采用TMT同位素标记法进行定量蛋白组学研究,以斑马鱼为模式生物,以课题组前期分离纯化地龙抗血栓活性蛋白DPf3为研究对象,预测地龙大分子活性成分抗血栓的作用机制,其潜在调控通路包括结氨基酸代谢、紧密连接、坏死性凋亡、吞噬、甘油磷酸代谢等。反映了地龙多成分、多靶点、多功效的特性,为其临床使用提供依据,为其抗血栓机制的进一步验证提供方向,并为采用现代药理学技术研究中药动物药提供参考。This study aims to explore the material basis and antithrombotic mechanisms of Dilong(Earthworms)based on network pharmacology and proteomics technology.The small molecular active components of Dilong(Earthworms)were firstly in-vestigated via the network pharmacology method,and a total of 101 chemical components were retrieved from literature to construct a composition database,including nucleotides,organic acids,amino acids and oligopeptides,lipids and others,and 798 of function-al targets were predicted.Based on the antithrombotic targets(fibrinogen,fibrin and plasmin)of Dilong(Earthworms)which have been verified by the research group in the earlier stage,1046 targets which have direct effects on Dilong(Earthworms)were mined through BioGRID database,which was a potential antithrombotic target library,with 81 targets in total.According to the protein-protein interaction networks of core targets and ingredients-targets-pathway network analysis,the results indicated that lipids,amino acids and oligopeptides might be the main antithrombotic compounds in Dilong(Earthworms),and fibronectin,plasmino-gen,mitogen-activated protein kinase-14,matrix metalloproteinases 9,epidermal growth factor receptor,prothrombin and fibrinogen might be the antithrombotic core targets of Dilong(Earthworms),and its antithrombotic effect was exhibited via regulating fo-cal adhesion,leukocyte transendothelial migrations,complement and coagulation,cell cycle and necroptosis.Then the antithrom-botic action mechanism of the macromolecular active component of Dilong(Earthworms)was predicted applying the TMT-labe-ling quantitative proteomics method using zebrafish as the experimental animal and the formerly purified antithrombotic protein DPf3 as the research object.The results indicated that the potential regulatory pathways included amino acid metabolism,tight junction,necroptosis,phagocytosis and glycerol phosphate metabolism.This study reflected the characteristics of multi-compo-nents,multi-targets and multi-functions of Dilong(Earthworms),

关 键 词：网络药理学 蛋白组学 地龙 抗血栓 作用机制 

分 类 号：R282.71[医药卫生—中药学]