计算机模拟研究肝素联合川芎嗪的协同抗血栓药理机制
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摘要
目的:基于计算机模拟研究肝素联合川芎嗪治疗血栓性疾病协同增效的分子机制。方法:在PDB数据库中寻找肝素靶蛋白,与肝素进行分子对接;利用反向分子对接服务器(DRAR-CPI)和毒性与基因比较数据库(CTD)对川芎嗪进行靶点预测和筛选;运用正向分子对接软件AutoDock Vina探究川芎嗪与潜在抗血栓靶点亲和力;利用STRING平台构建肝素及川芎嗪抗血栓靶点的蛋白互作(PPI)网络;利用生物学信息注释数据库(KEGG)将川芎嗪及肝素的抗血栓靶点进行KEGG通路富集。结果:肝素诱导ATⅢ变构、激活,抑制凝血酶活性。而川芎嗪则能抑制凝血因子Ⅹ、凝血因子IX、凝血因子VII、酪氨酸蛋白激酶JAK2、纤溶酶原激活物抑制剂1、血浆激肽释放酶6个靶点。二者联合从上下游多重阻断凝血级联反应通路。结论:肝素联合川芎嗪能作用于多个凝血因子以及凝血酶、纤溶酶原激活物抑制剂等血栓形成关键蛋白,调节凝血纤溶系统,协同产生抗凝、溶栓类抗血栓作用。
OBJECTIVE To study the synergistic mechanism of heparin and ligustrazine in the treatment of thrombotic disease by computer simulation.METHODS Searching the target of heparin in PDB Database and made molecular docking.Targetsof ligustrazine were predicted and screened relying on reverse molecular docking server(DRAR-CPI)and the Comparative Tox-icogenomics Database.AutoDock Vina was used to study the affinity between the ligustrazine and antithrombotic targets.Pro-tein-protein interactions(PPI)network of the antithrombotic targets of heparin and ligustrazine was structured based onSTRING platform.KEGG(Kyoto Encyclopedia of Genes and Genomes)pathways of the antithrombotic targets of heparin andligustrazine were analyzed based on biological information annotation databases DAVID(Database for Annotation,Visualizationand Integration Discovery).RESULTS Heparin induced AT Ⅲallosteric activation and inhibited thrombin.Ligustrazine actedon coagulation factorⅩcoagulation factorⅨ,tyrosine-protein kinase JAK2,coagulation factorⅦ,plasminogen activator in-hibitor 1,plasma kallikrein 6 targets.Heparin and ligustrazine realized the function of antithrombus by blocking multiply coag-ulation cascades pathway in the upstream and downstream.CONCLUSIONHeparin and ligustrazine can act on a number of co-agulation factors and thrombogenic key proteins such as thrombin and plasminogen activator inhibitors,regulate the coagulation and fibrinolytic system,and coproduce anticoagulant and thrombolytic antithrombotic effects.
OBJECTIVE To study the synergistic mechanism of heparin and ligustrazine in the treatment of thrombotic disease by computer simulation.METHODS Searching the target of heparin in PDB Database and made molecular docking.Targetsof ligustrazine were predicted and screened relying on reverse molecular docking server(DRAR-CPI)and the Comparative Tox-icogenomics Database.AutoDock Vina was used to study the affinity between the ligustrazine and antithrombotic targets.Pro-tein-protein interactions(PPI)network of the antithrombotic targets of heparin and ligustrazine was structured based onSTRING platform.KEGG(Kyoto Encyclopedia of Genes and Genomes)pathways of the antithrombotic targets of heparin andligustrazine were analyzed based on biological information annotation databases DAVID(Database for Annotation,Visualizationand Integration Discovery).RESULTS Heparin induced AT Ⅲallosteric activation and inhibited thrombin.Ligustrazine actedon coagulation factorⅩcoagulation factorⅨ,tyrosine-protein kinase JAK2,coagulation factorⅦ,plasminogen activator in-hibitor 1,plasma kallikrein 6 targets.Heparin and ligustrazine realized the function of antithrombus by blocking multiply coag-ulation cascades pathway in the upstream and downstream.CONCLUSIONHeparin and ligustrazine can act on a number of co-agulation factors and thrombogenic key proteins such as thrombin and plasminogen activator inhibitors,regulate the coagulation and fibrinolytic system,and coproduce anticoagulant and thrombolytic antithrombotic effects.
引文
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[22]Tang J,Yu C L,Williams S R,et al.Expression,crystallization,and three-dimensional structure of the catalytic domain of human plasma kallikrein[J].J Biol Chem,2005,280:41077-41089.
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[24]Meng M,Li JG,Wang HH,et al.Research Progress on Targeted Antithrombotic Drugs[J].Northern Pharm(北方药学),2016,13(8):104-107.
[25]Sun XQ.Advances in research on antithrombotic drugs[J].China Prac Med(中国实用医药),2010,5(26):241-242.
[26]Chen YH,Liu D,Peng LF,et al.Research progress on factor XI as a novel target for antithrombotic therapy[J].Chin Pharm Bull(中国药理学通报),2015,31(5):619-622.
[27]Shi DZ.Discussion on the pharmacological action of Ligustrazine[J].Chin J of Integr Trad West Med(中国中西医结合杂志),2003,23(5):377-378.
[28]Li QJ,Wang W,Ma HX,et al.Introduction to Antithrombin[J].Prog Vete Med(动物医学进展),2011,32(9):106-109.
[29]Chen HF.Design,Synthesis and cardiovascular activity studies of novel factor Xa inhibitors and ligustrazine derivatives[D].Ji Nan:Shandong university(济南:山东大学),2014
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[31]Yi ZZ,Huang WX,Xie ZD,et al.Ligustrazine on experimental acute pancreatitis rats plasma coagulation and fibrinolytic function[J].Hunan J TCM(湖南中医杂志),2012,28(1):96-98.
[32]Zhang ZJ.Kallikrein detection and its physiological effect[J].Chin Pharm Abstr(中国药学杂志),1994,29(4):195-198.
[33]Li D,Du YH,Li J.Research progress of the effects of Kallikrein-kinin system on cerebral infarction[J].J Tianjin Univ Trad Chin Med(天津中医药大学学报),2015,34(3):184-187.
[34]Huang P,Liu PB,Luo JW,et al.Effects of ulinastatin on coagulation in children after cardiopulmonary bypass[J].Chin JContemp Pediatr(中国当代儿科杂志),2012,14(4):279-281.
[35]Wang XP,Cheng JF,Zheng QQ,et al.Analysis for the coagulation disorder and complicated vascular diseases in patients with JAK2V617F mutation positive myeloproliferative neoplasms[J].Hebei Med J(河北医药),2016,38(11):1617-1620.
[36]Han ZY,Tian PF.Application of KEGG Database in Research of Biosynthesis[J].Biotech Bull(生物技术通报),2011,(1):76-82.
[2]Ji SL,Sang Q,Zhang TM.Research progress on source control,structure analysis and relationship between structure and biological activity of Heparin[J].Chin Pharm Abstr(中国药学杂志),2012,47(9):660-663.
[3]Chen LY,Yan LP,Li MF,et al.Adverse Reactions in Acute Coronary Syndrome Patients Treated with Low molecular weight Heparin Subcutaneously Injected in Different Methods[J].Chinese General Pratice(中国全科医学),2017,20(23):2911-2914.
[4]Zhou X,Wang YX,Miao ZL.TMP Combined with Low Molecular Weight Heparin Treatment of Unstable of Angina Medicine Randomized Parallel Group Study Salvia[J].J Prac Trad Chin Int Med(实用中医药杂志),2014,28(6):46-48.
[5]Zhao MX.Clinical study on the treatment of acute cerebral infarction with ligustrazine injection combined with low molecular weight Heparin[J].Prim Heal Care Chin(中国初级卫生保健),2013,5(31):77-78.
[6]Tian YJ.Clinical observation of low molecular weight Heparin combined with ligustrazine to prevent deep venous thrombosis after hip fracture[J].J Prac Trad Chin Int Med(实用中医药杂志),2017,9(33):1065-1066.
[7]He J,Gong YP,Wang BL,et al.Antithrombotic effect of Ligustrazine injection combined with aspirin and clopidogrel in rabbits[J].Chin Clin Pharmacol(中国临床药理学杂志),2017,33(7):612-615.
[8]Chen LW,Jia YJ.Ligustrazine hydrochloride combined with low dose of Heparin calcium for malignant treatment Clinical observation of tumor complicated with deep venous thrombosis of lower extremity[J].Shaanxi Trad Chin Med(陕西中医),2015,36(2):176-177.
[9]Hopkins A L.Network pharmacology[J].Nat Biotechnol,2007,25:1110-1111.
[10]Li S.Network target:a starting point for traditional Chinese medicine network pharmacology[J].Chin Pharm Abstr(中国中药杂志),2011,36(15):2017-2020.
[11]Xu HY,Huang LQ,Lu P,et al.Modern Research Thinking of Traditional Chinese Medicine Based on In-vivo ADMEProcess and Network Pharmacology[J].Chin Pharm Abstr(中国中药杂志),2012,37(2):142-145.
[12]Trott O,Olson A J.Auto Dock Vina:improving the speed and accuracy of docking with a new scoring function,efficient optimization,and multithreading[J].J Comput Chem,2010,31(2):455-461.
[13]Ru JL.Construction and Utilization of Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform[D].Xi’an:Northwest Agricultural and Forestry University(西安:西北农林科技大学),2015.
[14]Luo H,Chen J,Shi LM,et al.DRAR-CPI:a server foridentifying drug repositioning potential and adverse drugreactions via the chemical-protein interactome[J].Nucleic Acids Res,2011,39:W492-W498.
[15]Szklarczyk D,Franceschini A,Kuhn M,et al.The STRING database in 2011:functional interaction networks of proteins,globally integrated and scored[J].Nucleic Acids Research,2011,39(Databaseissue):D561-D568.
[16]Huang DW,Sherman BT,Lempicki RA.Systematic and integrative analysis of large gene lists using DAVID bioinformatics resources[J].Nature Protoc,2009,4(1):44-57.
[17]Schweinitz A,Steinmetzer T,Banke I J,et al.Design of novel and selective inhibitors of urokinase-type plasminogen activator with improved pharmacokinetic properties for use as antimetastatic agents[J].J Biol Chem,2004,279:33613-33622.
[18]Maignan S,Guilloteau J P,Pouzieux S,et al.Crystal structures of human factor Xa complexed with potent inhibitors[J].J Med Chem,2000,43:3226-3232.
[19]Hopfner K P,Lang A,Karcher A,et al.Coagulation factor IXa:the relaxed conformation of Tyr99blocks substrate binding[J].Structure Fold Des,1999,7:989-996.
[20]Lucet I S,Fantino E,Styles M,et al.The structural basis of Janus kinase 2inhibition by apotent and specific pan-Janus kinase inhibitor[J].Blood,2006,107:176-183.
[21]Schweitzer B A,Neumann W L,Rahman H K,et al.The structural basis of Janus kinase 2inhibition by apotent and specific pan-Janus kinase inhibitor[J].Bioorg Med Chem Lett,2005,15:3006-3011.
[22]Tang J,Yu C L,Williams S R,et al.Expression,crystallization,and three-dimensional structure of the catalytic domain of human plasma kallikrein[J].J Biol Chem,2005,280:41077-41089.
[23]Xue Y,Bjorquist P,Inghardt T,et al.Interfering with the inhibitory mechanism of serpins:crystal structure of a complex formed between cleaved plasminogen activator inhibitor type 1and a reactive-centre loop peptide[J].Structure,1998,6:627-636.
[24]Meng M,Li JG,Wang HH,et al.Research Progress on Targeted Antithrombotic Drugs[J].Northern Pharm(北方药学),2016,13(8):104-107.
[25]Sun XQ.Advances in research on antithrombotic drugs[J].China Prac Med(中国实用医药),2010,5(26):241-242.
[26]Chen YH,Liu D,Peng LF,et al.Research progress on factor XI as a novel target for antithrombotic therapy[J].Chin Pharm Bull(中国药理学通报),2015,31(5):619-622.
[27]Shi DZ.Discussion on the pharmacological action of Ligustrazine[J].Chin J of Integr Trad West Med(中国中西医结合杂志),2003,23(5):377-378.
[28]Li QJ,Wang W,Ma HX,et al.Introduction to Antithrombin[J].Prog Vete Med(动物医学进展),2011,32(9):106-109.
[29]Chen HF.Design,Synthesis and cardiovascular activity studies of novel factor Xa inhibitors and ligustrazine derivatives[D].Ji Nan:Shandong university(济南:山东大学),2014
[30]Ruan QR,Song JX,Deng ZD.Study on Mechanism of Ligustrazini Against Thrombosis[J].Chin J Arterioscler(中国动脉硬化杂志),1998,6(4):300.
[31]Yi ZZ,Huang WX,Xie ZD,et al.Ligustrazine on experimental acute pancreatitis rats plasma coagulation and fibrinolytic function[J].Hunan J TCM(湖南中医杂志),2012,28(1):96-98.
[32]Zhang ZJ.Kallikrein detection and its physiological effect[J].Chin Pharm Abstr(中国药学杂志),1994,29(4):195-198.
[33]Li D,Du YH,Li J.Research progress of the effects of Kallikrein-kinin system on cerebral infarction[J].J Tianjin Univ Trad Chin Med(天津中医药大学学报),2015,34(3):184-187.
[34]Huang P,Liu PB,Luo JW,et al.Effects of ulinastatin on coagulation in children after cardiopulmonary bypass[J].Chin JContemp Pediatr(中国当代儿科杂志),2012,14(4):279-281.
[35]Wang XP,Cheng JF,Zheng QQ,et al.Analysis for the coagulation disorder and complicated vascular diseases in patients with JAK2V617F mutation positive myeloproliferative neoplasms[J].Hebei Med J(河北医药),2016,38(11):1617-1620.
[36]Han ZY,Tian PF.Application of KEGG Database in Research of Biosynthesis[J].Biotech Bull(生物技术通报),2011,(1):76-82.