To investigate the effect of wheat gluten hydrolysate(WGH) rich in glutamine prepared through the enzymatic hydrolysis by trysin on small intestinal mucosal by methotrexate induced mucositis model in rats. The results showed that the WGH can significantly increase intestinal mucosa weight, villus height, crypt depth, protein and DNA content, decrease diamine oxidase level in plasma and increase glutathione content of small intestine mucosa. The WGH can efficiently alleviate the intestinal mucosal injury in rats.