In-vitro, ex-vivo, and in-silico assessment of the antioxidative potential of the leaf of the aromatic plant, Homalomena aromatica Schott

Background: Homalomena aromatica Schott is a medicinal plant traditionally used for therapeutic purposes and known to contain diverse bioactive phytochemicals. Despite its ethnomedicinal relevance, comprehensive evaluation of its antioxidant efficacy and underlying molecular mechanisms remains limited. Objective: This study investigated the phytochemical profile and antioxidative potential of H. aromatica extracts. Materials and Methods: Antioxidant activity was assessed by DPPH, ABTS•+, and superoxide anion scavenging assays, along with reducing power based on Fe³⁺ to Fe²⁺ conversion. Protective effects against oxidative damage were further evaluated by erythrocyte hemolysis and lipid peroxidation inhibition in the liver of mice. Phytochemical profiling of the methanolic extract was conducted using LC–HRMS. Network pharmacology analysis was performed to identify key molecular targets, followed by molecular docking to assess ligand–target interactions. Results: The methanolic extract exhibited the highest phenolic and flavonoid contents, corresponding to superior radical scavenging and reducing activities. In contrast, the aqueous extract demonstrated the greatest efficacy in preventing hemolysis and lipid peroxidation. LC–HRMS analysis of the methanolic extract of H. aromatica (HAME) identified 24 major secondary metabolites. Network pharmacology highlighted glycogen synthase kinase-3β (GSK3B) as a key molecular target of these compounds. Notably, molecular docking revealed that 2,6-dihydroxy-7-methoxy-1,1,4a-trimethyl-3,4,10,10a-tetrahydro-2H-phenanthren-9-one showed strong binding affinity (−8.0 kcal/mol) toward GSK3B. Conclusions: H. aromatica extracts, particularly the methanolic fraction, possess high antioxidative capacity attributed to diverse phytochemicals. Furthermore, the identification of a potential GSK3B inhibitor underscores the therapeutic promise of this plant in the management of oxidative stress–related disorders.