The dextran sulfate sodium (DSS) model of colitis has been commonly utilized in mice to assess novel treatments for ulcerative colitis. Recent studies have indicated that morphological and biochemical changes extend to the small intestine (SI). This study aimed to characterize histological and biochemical changes in the SI during DSS colitis in wild-type (WT) and DPIV knock-out (DPIV Γ/Γ ) mice treated with saline or the DPIV inhibitors, Ile-Pyrr-(2-CN) Γ TFA or Ile-Thia. Groups (n ΒΌ 10) of DPIV Γ/Γ and WT mice were orally gavaged twice daily with saline, Ile-Pyrr-(2-CN) Γ TFA or Ile-Thia. Mice consumed 2% DSS in drinking water for 6 days to induce colitis. Small intestinal tissue was assessed for histological changes, sucrase, and DPIV activity and neutrophil infiltration. Jejunal villus length was increased in all groups after 6 days DSS consumption ( P < 0.05). Jejunal DPIV activity was significantly lower by 35% in WT mice receiving Ile-Pyrr-(2-CN) Γ TFA compared to saline controls. Jejunal MPO activity was significantly increased in the WT ΓΎ saline and DPIV Γ/Γ ΓΎ saline groups following DSS consumption, compared to WT and DPIV Γ/Γ controls at day 0. Increased sucrase activity was apparent at day 0 in DPIV Γ/Γ compared to WT mice ( P < 0.05). We conclude that DSS-induced damage is not restricted to the colon, but also extends to the small intestine. Furthermore, reduced or absent DPIV activity resulted in functional adaptations to brush border enzyme activity. DPIV inhibitors are now a recognized therapy for type-II diabetes. The work presented here highlights the need to delineate any long-term effects of DPIV inhibitors on SI function, to further validate their safety and tolerability.