Antibiotic overuse has accelerated antimicrobial resistance (AMR), posing significant health and environmental risks. This study investigates the potential of Withania somnifera (Ashwagandha) in combating AMR in Pseudomonas aeruginosa, which exhibits resistance to certain aminoglycoside antibiotics. Disc diffusion assays across 14 antibiotics identified resistance exclusively to gentamicin. Phytochemicals from various parts of Ashwagandha were tested against resistant P. aeruginosa strains. Molecular docking of 68 phytochemicals against MexZ and aminoglycoside 3’-N-acetyltransferase proteins identified lead compounds, while molecular dynamics simulations of MexZ with withaferin A and withanolide B assessed the stability of protein-ligand complexes. The methanolic root extract proved most effective, producing a 34 mm inhibition zone. Molecular docking revealed binding affinities below –8.0 kcal mol-1 in multiple compounds, with key stability parameters from Molecular docking simulation supporting the potential for drug design. The findings highlight Ashwagandha root phytochemicals as promising candidates for overcoming antibiotic resistance in P. aeruginosa.