Exploring Molecular Mechanism of Thymoquinone via cMet and PDGFR for Accelerated Hemostasis: An Approach Through In...

Modulation of coagulation activity by thymoquinone (TQ), a 1,4-benzoquinone derivative of Nigella sativa, prompted the identification and validation of a molecular mechanism for hemostasis. Consequently, this research utilizes network pharmacology, molecular docking, and dynamics approaches to investigate the role of TQ for improved hemostasis. Initially, protein targets related to hemostasis and TQ were extracted, where the common targets were subjected to KEGG pathway enrichment and gene ontology analysis to determine the top pathways responsible for inducing hemostasis. The key protein targets from the identified pathways were selected for molecular docking and MM-GBSA calculations, where the targets with the highest absolute Glide docking and ΔG_bind scores were further considered for molecular dynamics simulation. The results indicated that TQ exerts potential effects on hemostasis by modulating cMet, PDGFR, and PI3KA proteins involved in the EGFR-TKIR pathway. The nontoxic concentration of TQ was determined using the MTT assay, which revealed a safe concentration (< 2.5 µM) of TQ for biological applications. In practice, TQ was found to induce the formation of stable fibrin clots in platelet-poor plasma, confirming its hemostatic potential. Finally, in vivo studies in a rat tail hemorrhage model confirmed its clotting activity, where modulation of cMet and PDGFR indicates the potential of TQ in inducing rapid hemostasis. Thus, this study integrated in silico, in vitro, and in vivo approaches to investigate the role of TQ in hemostasis through the EGFR-TKIR pathway, a mechanism that remains largely unexplored in blood clotting.