Reports on the effects of cholestyramine on small intestinal structure of rats have produced contradictory data about changes in mucosal histomorphometry, perhaps due to interacting effects of dietary composition. In order to identify effects of cholestyramine and diet on structure of the small intestines, 40 male rats were divided into 4 groups of 10 and fed 1 of each of the following diets for 1 month: standard diet, purified fiber-free diet, standard diet + 2% cholestyramine, or purified fiber-free diet + 2% cholestyramine. Serum concentrations of cholesterol and triglycerides were moderately increased in rats fed the purified fiber-free diet versus the standard diet. Neither total length nor weight of small intestine were affected by either diet or cholestyramine. Mucosal weight was affected by interactions between cholestyramine and diet, indicating that outcome depended upon modulating effects of both variables. Significant interactions were similarly detected among the variables of anatomic site, diet, and cholestyramine for many histomorphometric parameters of intestinal mucosa. Cholestyramine reduced total mucosal thickness in both jejunum and ileum and reduced villus height and villus:crypt ratio in the ileum.