In a rabbit model of chronic ileal inflammation, we previously demonstrated that coupled NaCl absorption was reduced because of an inhibition of Cl⁻/but not Na⁺/H⁺exchange on the brush-border membrane (BBM) of villus cells. In this study we determined the alterations in Na⁺-stimulated glucose [Na⁺-O-methyl-d-glucose (Na⁺-OMG)] absorption during chronic ileitis. Na⁺-OMG uptake was reduced in villus cells from the chronically inflamed ileum. Na⁺-K⁺-adenosinetriphosphatase (ATPase), which provides the favorable Na⁺ gradient for this cotransporter in intact cells, was found to be reduced also. However, in villus cell BBM vesicles from the inflamed ileum Na⁺-OMG uptake was reduced as well, suggesting an effect at the level of the cotransporter itself. Kinetic studies demonstrated that Na⁺-OMG uptake in the inflamed ileum was inhibited by a decrease in the maximal rate of uptake for OMG without a change in the affinity. Analysis of steady-state mRNA and immunoreactive protein levels of this cotransporter demonstrates reduced expression. Thus Na⁺-glucose cotransport was inhibited in the chronically inflamed ileum, and the inhibition was secondary to a decrease in the number of cotransporters and not solely secondary to an inhibition of Na⁺-K⁺-ATPase or altered affinity for glucose.