β-Sitosterol from Ipomoea carnea Jacq. As a promising anti-inflammatory agent: Evidence from in silico modeling and in vitro validation

Inflammation involves complex signaling networks that require multi-target therapeutic approaches. In this study, five phytoconstituents of Ipomoea carnea Jacq. (ferulic acid, vanillic acid, β-sitosterol, 1-triacontanol, and 2,5-dihydroxybenzoic acid) were investigated using integrated computational and in vitro methods. Network pharmacology identified key inflammatory targets, including TNF, IL6, IL1B, MAPK1, MAPK14, STAT3, and NFKB1, enriched in NF-κB, TNF, IL-17, JAK-STAT, NOD-like receptor, and FoxO signaling pathways. Molecular docking against the 3HA8 receptor revealed β-sitosterol as the top-ranked ligand with a docking score of −7.0. QuickProp analysis predicted high membrane permeability for β-sitosterol; however, its high lipophilicity, extremely poor aqueous solubility, and multiple property alerts indicated that dissolution and formulation limitations may restrict its actual oral exposure. The β-sitosterol-3HA8 complex was further assessed through a 500 ns molecular dynamics simulation, where RMSD, RMSF, protein-ligand contacts, PCA, FEL, and DCCM analyses indicated stable binding and favorable conformational behavior. Density functional theory calculations yielded a HOMO-LUMO energy gap of 6.90 eV, consistent with a chemically hard electronic structure and a comparatively low propensity for frontier-orbital excitation within the applied computational model. In vitro studies demonstrated concentration-dependent inhibition of protein denaturation and HRBC membrane stabilization, reaching 76.32 ± 1.35% and 82.47 ± 1.42%, respectively, at 300 μg/mL. Collectively, these findings suggest that β-sitosterol may contribute to the anti-inflammatory potential of I. carnea through multi-target pathway modulation, stable receptor interaction, and membrane-protective effects. Further experimental validation is required.