Protective Effects of the Ethanolic Leaf Extract of Ocimum sanctum L. on Collagen-Induced Human Platelet Aggregation and CCl4-Induced Hepatotoxicity in Rats: Molecular Docking Analysis of Platelet Activation Signaling Targets

Natural products represent an important source of bioactive compounds with therapeutic potential in the fields of thrombotic and oxidative stress-related disorders. This study focuses on identifying active compounds from the ethanolic extract of Ocimum sanctum L. leaves (EEOSL) and validating its antiplatelet and hepatoprotective activities. GC-MS and LC-MS (both positive and negative ionization modes) analyses identified 155 and 113 compounds, respectively, constituting several bioactive compounds. Collagen-induced human platelet activation (CIHPA) and carbon tetrachloride (CCl4)-induced hepatotoxicity were used as rat models to assess the activity of EEOSL. EEOSL (1–10 mg/mL) was shown to significantly and dose-dependently inhibit collagen-induced platelet aggregation. The extract also inhibited P-selectin expression, ATP release and intracellular Ca2+ mobilization, suggesting inhibition of major pathways involved in platelet activation. To understand the mechanisms, molecular docking was carried out with key platelet signaling molecules such as GPVI, SYK, PLCγ2, P2RY12, and PI3Kβ. Some phytoconstituents showed good binding affinities, with apigenin having the highest binding affinity for PI3Kβ (−8.01 kcal/mol). This binding was further validated by molecular dynamics simulations showing the formation of stable complexes. Intraperitoneal injection of CCl4 significantly increased the serum hepatic markers (SGOT, SGPT, LDH and SALP); EEOSL treatment significantly reduced these increases in vivo. The extract normalized antioxidant enzyme activities (catalase, SOD and glutathione peroxidase) and antioxidant isozyme patterns. Histopathological results revealed a significant level of protection against liver damage. Overall, the results showed that EEOSL has phytoconstituents that could provide strong antiplatelet and hepatoprotective properties, likely due to their ability to modulate platelet signaling pathways and enhance antioxidant defense mechanisms.