Enhanced cyclooxygenase pathway metabolite, PGE2, inhibits SGLT1 mediated glucose absorption in chronically inflamed intestinal villus cells. Lactoferrin (Lf), a multifunctional glycoprotein is absorbed by receptor‐mediated transcytosis has been shown to exert anti‐inflammatory effects. This study was designed to investigate whether Lf would prevent PGE2 inhibition of SGLT1 mediated glucose absorption in enterocytes. [3H]O‐Methyl‐Glucose uptake was measured in 10 days postconfluent rat intestinalepithelial cells (IEC‐6) grown on transwell plates. The cells used were treated with phlorizin, PGE2, AH6809, and Lf. Na‐dependent glucose uptake, phlorizin, and immunoblotting studies established the activity and apical membrane localization of SGLT1. PGE2 inhibited SGLT1 in both concentration and time dependent manner with an inhibitory constant Ki of 50.03 nM and antagonized by prostanoid receptor inhibitor. PGE2 did not alter Na+/K+‐ATPase activity. In contrast, qRT‐PCR and western blot analyses revealed that SGLT1 specific transcripts and protein expression level were decreased 3‐fold by PGE2 treatment. Kinetic studies demonstrated that PGE2 inhibited SGLT1 by reducing the cotransporter expression which is reversed by treatment with Lf. We speculate that Lf ameliorates PGE2 inhibition of SGLT1 mediated glucose absorption by interfering with the signal transduction pathway in IEC‐6 cells.