13516 10.30848/PJB2026-6(17) <p> <em>In silico</em> and <em>in vitro</em> studies on the synergistic effect of <em>Punica granatum </em>L. and <em>Nigella sativa</em> L. extracts in inhibiting the growth of methicillin-resistant <em>Staphylococcus aureus</em></p> Syeda Noor-Ul-Ain Abid, Amjad Hassan, Azhar Mehmood, Muhammad Huzafa, Abbas Hussain Shah and Jawad Ahmed <p> Antimicrobial resistance (AMR) is a major public health concern due to antibiotic misuse. The prevalence of drug resistance has drawn attention to the antibacterial qualities of plants and their metabolites. <em>Punica granatum </em>and <em>Nigella sativa</em> are well-known plant species for their bioactive antimicrobial phytochemicals. However, the current research does not provide enough insight into the specific interactions between these phytochemicals and bacterial target proteins. The present study employed molecular docking to investigate the likely interactions of bioactive phytochemicals from <em>P. granatum </em>and<em> N. sativa </em>with the known antibacterial target proteins to fill this critical missing link. The drug-likeliness of the phytochemicals was predicted by SwissADME. Among the tested phytochemicals, punicalin, a compound unique to <em>P. granatum</em>, exhibited the highest binding affinity (-19.4 kcal/mol) followed by dithymoquinone (-12.9 kcal/mol), derived from <em>N. sativa </em>for iron-regulated surface determinant protein A of <em>Staphylococcus aureus</em>. Nonetheless, SwissADME predicted more favorable drug-likeness properties for dithymoquinone than punicalin. To validate the predicted results, powdered samples of <em>N. sativa </em>and <em>P. granatum</em> were extracted separately in methanol, ethyl acetate, and distilled water. Analysis of Variance (ANOVA) followed by Tukey HSD test was calculated at p &le; 0.05 to measure statistically significant differences among the means. In a well diffusion assay, the combination of methanolic extracts of <em>P. granatum</em> and <em>N. sativa </em>showed significantly the highest value (2.71 cm) of bacterial growth inhibition zone compared to the rest of the extracts, whereas the lowest value of bacterial growth inhibition zone was shown by aqueous extract of <em>N. sativa </em>(0.39 cm). Similar results were recorded in the disc diffusion assay. These results indicate that combining <em>P. granatum</em> and <em>N. sativa </em>extracts leads to synergistic antimicrobial activity against <em>S. aureus </em>which may prove a promising strategy for combating AMR.</p> <p> &nbsp;</p> Published 2026 58 6 1385 1391