BACKGROUND/OBJECTIVES: Inflammatory responses are key pathological factors in various canine diseases, making the control of inflammatory responses vital for canine health. This study examined the anti-inflammatory effects of rutin on DH82 cells, a type of canine macrophage, against lipopolysaccharide (LPS)-induced inflammatory responses. MATERIALS/METHODS: The inflammatory RESULTS: Rutin inhibited the LPS-induced increase in the protein and gene levels of pro-inflammatory cytokines (interleukin [IL]-1β, IL-6, tumor necrosis factor-α), while anti-inflammatory cytokines (IL-10, transforming growth factor-β1) levels remained unchanged. Furthermore, rutin suppressed the LPS-induced activation of phosphorylated extracellular signal-regulated kinase, Jun N-terminal kinase, inhibitor of nuclear factor kappa B, and nuclear factor kappa B (NF-κB) in DH82 cells. CONCLUSION: Rutin exerts anti-inflammatory effects by inhibiting the mitogen-activated protein kinase-NF-κB signaling pathway and reducing the production of pro-inflammatory cytokines in DH82 cells.