title: Insulin targets the Na(+)/K(+) ATPase in enterocytes via PI3K, PKC, and MAPKS.
abstract: The effect of insulin on intestinal Na(+)/K(+) ATPase is till now undetermined, and it is still unclear whether insulin exerts any modulatory effect on glucose absorption by targeting the ATPase. This work attempted to address this question and to unravel the signaling pathway involved using Caco-2 cells as a model. After an overnight starvation, cells were treated with insulin in presence and absence of specific inhibitors of some known mediators. The activity of the pump was assayed by measuring the ouabain-inhibitable inorganic phosphate (P(i)) released, whereas changes in its abundance were determined by western blot analysis. Insulin decreased the activity and abundance of the ATPase in a crude membrane homogenate. This effect disappeared completely upon inhibition of either phosphotidylinositol-3 kinase (PI3K) or protein kinase C (PKC), but was partially abolished when p38MAPK or MEK/ERK were inhibited separately. Activation of PKC with phorbol-12-myristate-13-acetate (PMA) imitated the effect of insulin and was not affected by inhibition of PI3K. The data suggest that PI3K and PKC are along the same pathway that branches into two separate ones involving each either p38MAP kinase or MEK/ERK. This hypothesis was confirmed by the data obtained from the treatment of Caco-2 cells with PMA, when p38MAPK and MEK/ERK were inhibited simultaneously. Concomitant inhibition of p38MAPK and MEK/ERK abrogated fully the effect of insulin, indicating that no other pathways are present in addition to the ones proposed above.
authors.affiliation: Department of Biology, American University of Beirut, Lebanon.
authors.forename: Maya F
authors.initials: MF
authors.lastname: Serhan
authors.forename: Sawsan I
authors.initials: SI
authors.lastname: Kreydiyyeh
mesh_terms: D000730:Androstadienes / Q000494:pharmacology; D018938:Caco-2 Cells; D004305:Dose-Response Relationship, Drug; D020895:Enterocytes / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism*; D004791:Enzyme Inhibitors / Q000378:metabolism; D005419:Flavonoids / Q000494:pharmacology; D006801:Humans; D007093:Imidazoles / Q000494:pharmacology; D007328:Insulin / Q000494:pharmacology*; D020928:Mitogen-Activated Protein Kinases / Q000378:metabolism*; D009281:Naphthalenes / Q000494:pharmacology; D019869:Phosphatidylinositol 3-Kinases / Q000378:metabolism*; D011493:Protein Kinase C / Q000378:metabolism*; D011725:Pyridines / Q000494:pharmacology; D015398:Signal Transduction / Q000187:drug effects / Q000502:physiology; D000254:Sodium-Potassium-Exchanging ATPase / Q000378:metabolism*; D000077191:Wortmannin
chemical_list: D000730:Androstadienes; D004791:Enzyme Inhibitors; D005419:Flavonoids; D007093:Imidazoles; D007328:Insulin; D020928:Mitogen-Activated Protein Kinases; D009281:Naphthalenes; D019869:Phosphatidylinositol 3-Kinases; D011493:Protein Kinase C; D011725:Pyridines; D000254:Sodium-Potassium-Exchanging ATPase; D000077191:Wortmannin; C058819:calphostin C; C090942:4-(4-fluorophenyl)-2-(4-hydroxyphenyl)-5-(4-pyridyl)imidazole; C093973:2-(2-amino-3-methoxyphenyl)-4H-1-benzopyran-4-one