In order to explore the protective effect of corn protein hydrolysate on intestinal barrier function, corn gluten meal was used as raw material and Alcalase was used as catalyst for enzymatic hydrolysis in current study. The effects of different enzymatic hydrolysis conditions on the content of glutamine in the hydrolysate were investigated, and the optimum hydrolysis system of corn protein was established. Then the sodium dextran sulfate was used as inducer to construct colitis model, and the alleviating effect of corn protein hydrolysate on colitis in mice was systematically evaluated. The results showed that the optimum hydrolysis conditions for Alcalase were a temperature of 64 ℃, hydrolysis time of 2.5 h, amount of enzyme of 1300 U/g. Under such condition, the content of glutamine in the hydrolysate could reach 8.79%. Corn protein hydrolysate (375 and 625 mg/kg·bw) significantly improved weight loss in colitis mice, reduced the colonic shortening and the disease activity index, diminished the infiltration of inflammatory cells in the colonic tissue, regulated the level of inflammatory cytokines, and played a good role in relieving colitis in mice. This study would provide theoretical basis for expanding high-value utilization of corn gluten meal and developing functional food for intestinal protection.