Gout is a type of inflammatory arthritis characterized by monosodium urate (MSU) crystal-induced inflammation in joints and surrounding tissues. Recent research suggests that regular exercise has anti-inflammatory effects which may help reduce inflammation in chronic inflammatory diseases. Using NF-κB-luc reporter mice, we investigated the effects of exercise intensity on inflammation in an acute model of MSU-induced gout. Mice were exercised daily at low, medium, or high-intensities on a treadmill for 2 weeks before receiving intra-articular MSU injections. Assessment of NF-κB activity via in vitro imaging system (IVIS) revealed significant reductions in NF-κB activity in mice exercised at low and moderate-intensities compared to the high-intensity group or non-exercised control. Immunohistochemistry (IHC) of the macrophage marker F4/80, the granulocyte marker myeloperoxidase (MPO), and the inflammatory cytokine IL-1β similarly revealed significantly reduced macrophages/granulocytes and IL-1β at the site of MSU injection in the low and moderate-intensity groups. Toll-like receptor 2 (TLR2), an important mediator of the MSU response, was significantly decreased on peripheral neutrophils from the low and moderate-intensity groups via flow cytometry. The neutrophil chemokine CXCL1 was also significantly decreased in the exercised groups. An examination of IL-1β production in peripheral monocytes and neutrophils isolated from exercised mice and stimulated by MSU/LPS in vitro revealed increased intracellular levels of pro-IL-1β and significantly decreased IL-1β in the supernatant of neutrophils. Cumulatively, these results suggest that low to moderate-intensity exercise produces a tolerogenic effect which makes immune cells less responsive to inflammatory stimuli and may partially explain the underlying anti-inflammatory mechanisms of physical activity.