Network Pharmacology Analysis on the Anticoagulant Effect of the Chemical Constituents of Paeoniae Radix Rubra on Blood Stasis Syndrome in Rats

Aims: The aim of the study was to investigate the pharmacological role of different chemical constituents of Paeoniae Radix Rubra (PRR) in the modulation of anticoagulation in Blood Stasis Syndrome (BSS). Background: In Traditional Chinese Medicine (TCM), BSS refers to impaired circulation or stagnation of blood flow and formation of bruises. The primary therapeutic strategy to treat BSS involves invigorating blood circulation. PRR is a widely used TCM herb for treating acute and critical diseases caused by BSS. However, the anticoagulant effects of different compounds of PRR on BSS remain elusive. Objective: This study aimed to analyze the therapeutic effect of PRR on BSS and to assess the ameliorative effect of different chemical constituents of PRR on blood circulation, clotting time, and platelet aggregation in rats with acute BSS. Methods: TCMSP (Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform) was used to screen the targets of PRR, and genes causing BSS were predicted using PharmGKB, OMIM, and TTD databases. Intersection genes between PRR and BSS targets were visualized in Venn diagrams. Core target networks of Protein-Protein Interaction (PPI) and crosstargets were constructed using Cytoscape 3.7.1, and the cross-targets were enriched using the DAVID (Database for Annotation, Visualization, and Integrated Discovery) database. Furthermore, the effects of PRR on platelet aggregation, plasma viscosity, and whole blood viscosity in the rats with BSS were examined by blood rheology and other methods. The serum levels of Endothelin-1 (ET- 1), Nitric Oxide (NO), Thromboxaneb2 (TXB2), and 6-keto-Prostaglandin F1 α (6-keto-PGF1α) in the rats were measured by the ELISA (Enzyme-Linked Immunosorbent Assay) method. Results: The main active compounds of PRR, including total glycosides, flavonoids, and polysaccharides, were identified using the TCMSP database. A total of 31 cross-targets were obtained from the intersection between 129 active targets of PRR and 345 causative genes of BSS. PPI network identified genes such as ALB (Albumin), SRC (SRC Proto-Oncogene, Non-Receptor Tyrosine Kinase), and AKT1 (AKT Serine/Threonine Kinase 1) as the core targets of PRR in alleviating BSS. Enrichment analysis showed that the common targets were mainly associated with several biological processes, including lipid and atherosclerosis, adherens junction, and focal adhesion. Following the intervention with PRR extract, the whole blood viscosity and plasma viscosity were reduced, and platelet aggregation was inhibited in the model rats in comparison to the model group. Moreover, PRR treatment also promoted thrombin time (TT), prothrombin time (PT), and Activated Partial Thromboplastin Time (APTT), increased the level of NO and 6-keto-PGF1α, but reduced the level of Fibrinogen content (FIB) and ET-1 and TXB2 in the serum of the model rats. Discussion: The present research systematically explored the anticoagulant effect of the chemical constituents of Paeoniae Radix Rubra on blood stasis syndrome in rats, applying network pharmacology analysis. Conclusion: The current findings provided a theoretical foundation for the pharmacological basis of using PRR in the management of BSS.