Induction of colitis by 2,4,6-Trinitrobenzenesulphonic acid (TNB) in the rat is a widely used experimental model of inflammatory bowel disease. Action of TNB as a hapten, induces colitis involving infiltration of colonic mucosa by neutrophils and macrophages and increased production of inflammatory mediators. The aim of the present study was to measure nitric oxide synthase (NOS) activity and characterize relations between inducible NOS (iNOS) activity and other signs of inflammation in TNB-induced colitis. A profound and sustained increase in the activity of iNOS was found in the colon. The activity of NOS in the spleen was also increased, but remained at low levels as compared to those in colon. No increases in plasma nitrite + nitrate concentrations were found suggesting local rather than systemic induction of iNOS. The increase in iNOS activity in the colon was preceded by macroscopic inflammatory lesions, like hyperemia, ulcerations and edema formation as well as neutrophil accumulation in the gastric mucosa and increased circulating concentrations of PGE2 metabolite (PGEM). Concentrations of PGEM in the plasma and myeloperoxidase activity (MPO; marker of neutrophil infiltration) in the gut declined in 48h whereas increased iNOS activity and the macroscopic inflammatory lesions remained over the 72h follow-up period. The results demonstrate increased local iNOS activity in TNB-Induced colitis mimicking the situation in human inflammatory bowel disease.