Theoretical exploring of potential mechanisms of antithrombotic ingredients in danshen-chishao herb-pair by network pharmacological study, molecular docking and zebrafish models

• Published in: Chinese medicine Vol. 19; no. 1; pp. 1 - 16
• Main Authors: Rao, Chang, Hu, Ruixue, Hu, Yongxin, Jiang, Yan, Zou, Xu, Tang, Huilan, Chen, Xing, He, Xiaoli, Hu, Guang
• Format: Journal Article
• Language: English
• Published: London BioMed Central Ltd 16.07.2024; BioMed Central; BMC
• Subjects: Acids; Albumin; Anticoagulants; Arteriosclerosis; Aspirin; Atherosclerosis; Blood clot; Cardiovascular disease; Caspase-3; Coagulation factor Xa; Danio rerio; Embryos; Epidermal growth factor receptors; Fibroblast growth factor 2; Gelatinase A; Gelatinase B; Gene expression; Growth factors; Health aspects; Herbal medicine; High performance liquid chromatography; Lipid metabolism; Matrix metalloproteinase; Medicine, Chinese; Metalloproteinase; Metalloproteins; Molecular modelling; Network pharmacology; Paeoniflorin; Protein kinases; Protein-protein interactions; Radix Paeoniae Rubra; Reagents; RNA; Salvia miltiorrhiza; Salvianolic acid A; Src protein; Thrombin; Thrombosis; Traditional Chinese medicine; Vascular endothelial growth factor; Vascular endothelial growth factor receptor 2; Zebrafish; China; Beijing China
• ISSN: 1749-8546; 1749-8546
• DOI: 10.1186/s13020-024-00970-6

Background Salvia miltiorrhiza (Danshen, DS) and Radix Paeoniae Rubra (Chishao, CS) herbal pair (DS-CS) is a famous traditional Chinese combination which has been used as antithrombotic formular for centuries. However, there is still lack of sufficient scientific evidence to illustrate its underlying mechanisms. The purpose of this study is to investigate the antithrombotic effects of DS-CS extract in zebrafish and explore its possible mechanism of action. Methods The quality of traditional Chinese medicines DS and CS granules was evaluated using High Performance Liquid Chromatography (HPLC). Subsequently, the therapeutic effect of the DS-CS combination and its components, Salvianolic Acid A (SAA) and Paeoniflorin (PF), in various concentrations on thrombosis was experimentally validated. Moreover, the interaction between DS-CS and the thrombosis disease targets was analyzed through network pharmacology, predicting the potential antithrombotic mechanism of DS-CS. Molecular docking and in vivo zebrafish experiments were conducted to validate the predicted targets, with qRT-PCR utilized for target validation. Results DS-CS exhibited anti-thrombotic effect in zebrafish with concentrations ranging from 25 to 300 [mu]g/mL. The co-administration of PF and SAA at 25 [mu]g/mL each revealed a synergistic antithrombotic effect exceeding that of individual components when contrasted with PHZ treatment. Protein-protein interaction (PPI) analysis identified key genes, including Albumin (ALB), Proto-oncogene tyro-sine-protein kinase Src (SRC), Matrix metalloproteinase-9 (MMP9), Caspase-3 (CASP3), Epidermal growth factor receptor (EGFR), Fibroblast growth factor 2 (FGF2), Vascular endothelial growth factor receptor 2 (KDR), Matrix metalloprotein-ase-2(MMP2), Thrombin (F2), and Coagulation factor Xa (F10), associated with the antithrombotic action of PF and SAA. Furthermore, KEGG pathway analysis indicated involvement of lipid metabolism and atherosclerosis pathways. Molecular docking revealed strong binding of PF and SAA to pivotal hub genes, such as SRC, EGFR, and F10. The experimental findings demonstrated that DS-CS could upregulate the mRNA expression levels of EGFR while inhibiting F10 and SRC mRNA levels, thereby ameliorating thrombotic conditions. Conclusion This research provided valuable insights into the potential mechanisms underlying the antithrombotic activity of DS-CS. Our findings suggested that PF and SAA could be the key active ingredients responsible for this activity. The antithrombotic effects of DS-CS appeared to be mediated through the regulation of mRNA expression of SRC, EGFR, and F10. These results enhanced our understanding of DS-CS's therapeutic potential and lay the groundwork for future studies to further elucidate its mechanisms of action. Keywords: Salvia miltiorrhiza, Radix Paeoniae Rubra, Salvianolic acid A, Paeoniflorin, Zebrafish, Network pharmacology, Molecular docking, Antithrombotic effect