Therapeutic potential of Xylocarpus granatum bark extracts: antioxidant, anti-inflammatory, cytotoxic and molecular insights

Free radicals contribute to various diseases, while antioxidants, like phenolic compounds and flavonoids, counteract their effects. This study examines the antioxidant, anti-inflammatory, and cytotoxic properties of Xylocarpus granatum bark extracts, emphasizing ethyl acetate fraction (EAF) and the isolated compound from EAF. The antioxidant constituents like total phenolic and flavonoid contents were quantified and antioxidant activity was evaluated using 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging assays. The cytotoxic potential of the extracts and the isolated compound was evaluated against brine shrimp and the MCF-7 breast cancer cell line using the MTT assay. Anti-inflammatory effects were evaluated using heat-induced hemolysis and RBC membrane stabilization assays to assess membrane-stabilizing and anti-inflammatory activity. Molecular docking studies were performed using PyRx and discovery studio. EAF demonstrated the highest total phenolic content (124.56 ± 1.35 mg GAE/g) and total flavonoid content (24.69 ± 0.48 mg QE/g), with superior performance in DPPH assays with IC₅₀ values of 3.84 µg/ml and that of pure compound, (+) catechin was 3.89 µg/ml. Anti-inflammatory effects revealed strong activity, with EAF and pure compound showing inhibition rates comparable to standard drugs like diclofenac-Na. Cytotoxicity, assessed through brine shrimp lethality and MCF-7 breast cancer cell line assays, highlighted EAF's potent activity (LC₅₀: 24.15 µg/ml) antiproliferative efficacy in a dose-dependent manner. Molecular docking studies confirmed (+) catechin's dual role as a COX-2/Caspase 9 inhibitor, elucidating its potential mechanisms of action. These findings underscore the therapeutic potential of Xylocarpus granatum extracts, particularly EAF and the isolated compound, in managing oxidative stress, inflammation, and cancer progression.