Scientific Validation of the Traditional Anti-Inflammatory Use of the Sapindus mukorossi Seed Kernel: In Vitro and In Silico Insights

S. mukorossi ., commonly known as soapnut, is a traditional medicinal plant widely used in Asian ethnomedicine for treating infections and inflammatory disorders. However, systematic validation of its pharmacological potential remains limited. The antimicrobial and anti-inflammatory properties of S. mukorossi extract were evaluated using in vitro and biochemical assays. Cytotoxicity was assessed in RAW 264.7 macrophages using the MTT assay. Antibacterial activity was determined by agar well diffusion and minimum inhibitory concentration (MIC) assays against Escherichia coli, Klebsiella pneumoniae , and Staphylococcus aureus , while antifungal efficacy was tested against Candida albicans using disc diffusion. Anti-inflammatory potential was examined by measuring TNF-α levels through ELISA and cyclooxygenase-2 (COX-2) inhibition via a colorimetric assay. Molecular docking and molecular dynamics simulations, along with gmx_MMPBSA free energy calculations, were performed to assess the binding affinity of identified compounds (isosilybin A, orientin) with IRAK4. The extract demonstrated significant antimicrobial activity, producing notable inhibition zones against both Gram-positive and Gram-negative bacteria, with low MIC values indicating potent bacteriostatic effects. Antifungal activity against C. albicans was comparable to amphotericin B. The extract exhibited low cytotoxicity toward RAW 264.7 cells and effectively suppressed inflammatory mediators, showing dose-dependent inhibition of COX-2 activity and a marked reduction in TNF-α secretion. In silico analyses revealed stable binding of isosilybin A and orientin to IRAK4, as confirmed by molecular dynamics and gmx_MMPBSA free energy calculations (ΔGbind up to − 47.8 kcal/mol). The findings substantiate the traditional use of S. mukorossi as a natural antimicrobial and anti-inflammatory agent. Its bioactive constituents may serve as potential leads for developing plant-derived therapeutic agents targeting infectious and inflammatory diseases. Further phytochemical characterization and molecular mechanism studies are warranted to elucidate the active principles and their targets.