Intestinal glucose uptake (GUi) from blood increased when blood flow (BF) was increased. The increase in BF could elevate shear stress. Therefore, we hypothesize that shear stress-induced release of autacoids mediates the increase in GU_i. A surgically separated segment of small intestine was perfused in situ with the use of an arterial circuit in anesthetized cats. Arterial and portal blood samples were taken simultaneously for assessment of GU_i. Adenosine was used to elevate intestinal BF. The GU_i increased by 45.0 ± 18.3 from 25.3 ± 3.8 µmol · min¹ · 100 g tissue¹ when the BF increased about four times. It was not a direct effect of adenosine because GU_i was not altered if the flow was held constant. This increase was blocked by a cyclooxygenase inhibitor, indomethacin, but not by nitric oxide synthase blocker N^G-nitro-L-arginine methyl ester. Furthermore, prostaglandin F₂ (PGF₂) but not PGE₂ or PGI₂ reversed the blockade of the increase in GU_i after indomethacin during elevated blood flow, whereas they had no influence on basal uptake. The results suggest that shear stress-induced release of PGF₂ mediated the increase in GU_i when blood flow was elevated.