本研究基于谱效关系-整合分子对接,探索天花粉及其壳核抗凝血的物质基础和作用机理。建立了天花粉及其壳和仁的高效液相色谱（HPLC）指纹图谱。低剂量和高剂量小鼠的凝血酶原时间（PT）和活化部分凝血活酶时间（APTT）（5，30g·kg〜（-1），分别)天花粉精液，贝壳,和内核组被确定为凝血标志物。采用邓氏关联度均值计算方法(MATLAB)对天花粉及其壳核的谱-效应关系和抗凝物质基础进行了分析,得到了常见的有效成分聚类。然后从TCMSP中检索组分簇和凝血的共同靶标，瑞士目标预测，GenCLiP3,GeneCards,和大卫,其次是基因本体论（GO）术语富集和京都基因和基因组百科全书（KEGG）途径富集的目标。SYBYL-X2.1.1验证了组分簇的主要抗凝血分子机制。天花粉及其壳核的谱效关系与剂量呈正相关。每种成分对抗凝的贡献并不相同，表明抗凝的物质基础是不同的，但它们具有共同的有效成分(即共同的物质基础)，如腺嘌呤（峰3），尿嘧啶（峰值4），次黄嘌呤（峰6），黄嘌呤（峰9），和腺苷(峰11)。网络药理学表明，这些成分可以作用于多种靶蛋白，如NOS3，KDR，和PTGS2,并通过多种途径如VEGF信号通路发挥抗凝作用。它们涉及蛋白质水解等生物学功能，细胞成分，如细胞质，和分子功能。分子对接结果表明,这些组分与NOS3(PDBID:1D0C)的结合自由能,KDR(PDBID:5AMN),PTGS2(PDBID:4COX)≤-5kJ·mol~(-1),对接构象稳定。光谱-效应关系-整合分子对接可用于优化，虚拟筛选,复杂的中药化学和生物信息的验证。天花粉及其壳核具有共同的抗凝物质基础,通过多种靶点和途径发挥抗凝作用。
This
study explored the anticoagulant material basis and mechanism of Trichosanthis Semen and its shell and kernel based on spectrum-effect relationship-integrated molecular docking. High performance liquid chromatography(HPLC) fingerprints of Trichosanthis Semen and its shell and kernel were established. Prothrombin time(PT) and activated partial thromboplastin time(APTT) in mice in the low-and high-dose(5, 30 g·kg~(-1), respectively) Trichosanthis Semen, the shell, and kernel groups were determined as the coagulation markers. The spectrum-effect relationship and anticoagulant material basis of Trichosanthis Semen and its shell and kernel were analyzed with mean value calculation method of Deng\'s correlation degree(MATLAB) and the common effective component cluster was obtained. Then the common targets of the component cluster and coagulation were retrieved from TCMSP, Swiss-TargetPrediction, GenCLiP3, GeneCards, and DAVID, followed by Gene Ontology(GO) term enrichment and Kyoto Encyclopedia of Genes and Genomes(KEGG) pathway enrichment of the targets. The main anticoagulant molecular mechanism of the component cluster was verified by SYBYL-X 2.1.1. The spectrum-effect relationship of Trichosanthis Semen and its shell and kernel was in positive correlation with the dosage. The contribution of each component to anticoagulation was not the same, suggesting that the material basis for anticoagulation was different, but they have common effective components(i.e. common material basis), such as adenine(peak 3), uracil(peak 4), hypoxanthine(peak 6), xanthine(peak 9), and adenosine(peak 11). Network pharmacology showed that these components can act on multiple target proteins such as NOS3, KDR, and PTGS2, and exert anticoagulant effect through multiple pathways such as VEGF signaling pathway. They involved the biological functions such as proteolysis, cell component such as cytosol, and molecular functions. The results of molecular docking showed that the binding free energy of these components with NOS3(PDB ID: 1 D0 C), KDR(PDB ID: 5 AMN), and PTGS2(PDB ID: 4 COX) was ≤-5 kJ·mol~(-1), and the docking conformations were stable. Spectrum-effect relationship-integrated molecular docking can be used for the optimization, virtual screening, and verification of complex chemical and biological information of Chinese medicine. Trichosanthis Semen and its shell and kernel have the common material basis for anticoagulation and they exert the anticoagulant through multiple targets and pathways.
