The abuse of antibiotics leads to the increasingly serious problem of bacterial drug resistance, which makes an urgent tendency to find promising alternative drugs for antibiotics. Paeonol, as a natural plant active ingredient, exhibits good antibacterial potential. To further study the molecular mechanism of the antibacterial effect of paeonol, the potential antibacterial mechanism of paeonol was analyzed by using proteomics and bioinformatics with Aeromonas hydrophila as the main experimental object.