BACKGROUND: Loss of mucosal integrity is associated with intestinal hyperpermeability, which may be inhibited by glutamine. The nature of this effect is unknown. The effect of glutamine on protein kinase C (PKC)-mediated hyperpermeability in HT-29Cl.19A intestinal cells was studied. METHODS: Confluent monolayers of HT-29C1.19A cells were cultured on permeable filters and mounted in Ussing chambers for permeability studies. Apical to basolateral transepithelial permeability for intact horseradish peroxidase (HRP) was determined. Phorbol-12,13-dibutyrate (PDB) was used to activate PKC-mediated hyperpermeability, and the effect of glutamine (0.6 mmol/L) was studied. RESULTS: Two hours of PDB stimulation increased the HRP flux, reaching five times control values after 4 hours. Bilateral exposure to glutamine for 4 hours reduced PDB-induced hyperpermeability (37%). Preincubation with glutamine 2 hours before PDB stimulation showed an earlier and greater effect (3 hours, 43%; 4 hours, 50%). This bilateral effect of glutamine was mimicked by separate apical exposure. Basolateral exposure alone had no effect. CONCLUSIONS: Glutamine rapidly reduced the PKC-mediated hyperpermeability for HRP in HT-29Cl.19A intestinal cells. The dependency on apical exposure suggests that glutamine may be more effective when delivered by the enteral route.
BACKGROUND: Loss of mucosal integrity is associated with intestinal hyperpermeability, which may be inhibited by glutamine. The nature of this effect is unknown. The effect of glutamine on protein kinase C (PKC)-mediated hyperpermeability in HT-29Cl.19A intestinal cells was studied. METHODS: Confluent monolayers of HT-29C1.19A cells were cultured on permeable filters and mounted in Ussing chambers for permeability studies. Apical to basolateral transepithelial permeability for intact horseradish peroxidase (HRP) was determined. Phorbol-12,13-dibutyrate (PDB) was used to activate PKC-mediated hyperpermeability, and the effect of glutamine (0.6 mmol/L) was studied. RESULTS: Two hours of PDB stimulation increased the HRP flux, reaching five times control values after 4 hours. Bilateral exposure to glutamine for 4 hours reduced PDB-induced hyperpermeability (37%). Preincubation with glutamine 2 hours before PDB stimulation showed an earlier and greater effect (3 hours, 43%; 4 hours, 50%). This bilateral effect of glutamine was mimicked by separate apical exposure. Basolateral exposure alone had no effect. CONCLUSIONS:Glutamine rapidly reduced the PKC-mediated hyperpermeability for HRP in HT-29Cl.19A intestinal cells. The dependency on apical exposure suggests that glutamine may be more effective when delivered by the enteral route.
Authors: Mechteld A R Vermeulen; Jeffrey de Jong; Mathijs J Vaessen; Paul Am van Leeuwen; Alexander P J Houdijk Journal: World J Gastroenterol Date: 2011-03-28 Impact factor: 5.742