计算机模拟研究肝素联合川芎嗪的协同抗血栓药理机制

Synergistic antithrombotic mechanism of heparin and ligustrazine by computer simulation study

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.