Hemostatic effects of combined Thymus vulgaris and Medicago sativa extracts: in silico and in vivo study.

Objective: This study aims to evaluate the hemostatic effects of combined extracts from Thymus vulgaris and Medicago sativa in an animal model, focusing on their impact on coagulation indices. We hypothesize that the combined extracts will modulate the extrinsic and intrinsic coagulation pathways, improving hemostasis.

Methods: Thirty-two male NMRI mice were randomly assigned to four groups (n = 8): negative control (distilled water), 300 mg/kg/day M. sativa (MS300), 100 mg/kg/day T. vulgaris extract (TV100), and combined extracts (TV100 + MS300). After 14 days of treatment, blood samples were collected to measure prothrombin time (PT) and activated partial thromboplastin time (aPTT). Chemical analysis identified active compounds, and molecular docking studies were performed to assess their interaction with coagulation factor VIIa (FVIIa).

Results: The treated groups showed significant changes in coagulation indices compared to the control. PT was significantly decreased (P < 0.05), and aPTT was significantly increased (P < 0.05) in the M. sativa, T. vulgaris, and combined extract groups. The combined extract showed the most significant effect. Computational analyses revealed that compounds like Scandenon and Vitamin E interacted with FVIIa, suggesting their role in modulating the extrinsic coagulation pathway. These compounds showed strong binding affinity to FVIIa.

Conclusion: Combined extracts of T. vulgaris and M. sativa significantly influence coagulation, especially the extrinsic pathway. The presence of aromatic, hydroxyl, alcoholic, and phenolic groups in these compounds likely contributes to their interaction with coagulation factors. These findings support the potential development of plant-based hemostatic agents for clinical use.