Curcumin is known to possess a wide range of pharmacological activities for the treatment of chronic or inflammatory diseases, Alzheimer's disease, and various cancers. However, the therapeutic efficacy of curcumin is restricted by its poor bioavailability after oral administration. In this study, the effects of various cyclodextrins on the intestinal absorption of curcumin were evaluated in rat intestine by an in situ closed-loop method. Among the tested cyclodextrins, 50 mM α-cyclodextrin significantly enhanced the absorption of curcumin without inducing any intestinal toxicity. The analysis of cellular transport across Caco-2 cell monolayers showed that 50 mM α-cyclodextrin reduced the transepithelial electrical resistance value of cell monolayers and improved the permeability of 5(6)-carboxyfluorescein, a poorly absorbable drug, which is mainly transported via a paracellular pathway. Furthermore, the western blotting analysis showed that α-cyclodextrin decreased the expression of claudin-4, a tight junction-associated protein, in brush border membrane vesicles. Additionally, α-cyclodextrin increased the membrane fluidity of lipid bilayers in brush border membrane vesicles and may also have promoted the permeation of drug molecules via a transcellular pathway. These results suggested that α-cyclodextrin might enhance the intestinal absorption of curcumin via both paracellular and transcellular pathways.