title: Regulation of ENaC and CFTR expression with K+ channel modulators and effect on fluid absorption across alveolar epithelial cells.
abstract: In a recent study (Leroy C, Dagenais A, Berthiaume Y, and Brochiero E. Am J Physiol Lung Cell Mol Physiol 286: L1027-L1037, 2004), we identified an ATP-sensitive K(+) (K(ATP)) channel in alveolar epithelial cells, formed by inwardly rectifying K(+) channel Kir6.1/sulfonylurea receptor (SUR)2B subunits. We found that short applications of K(ATP), voltage-dependent K(+) channel KvLQT1, and calcium-activated K(+) (K(Ca)) channel modulators modified Na(+) and Cl(-) currents in alveolar monolayers. In addition, it was shown previously that a K(ATP) opener increased alveolar liquid clearance in human lungs by a mechanism possibly related to epithelial sodium channels (ENaC). We therefore hypothesized that prolonged treatment with K(+) channel modulators could induce a sustained regulation of ENaC activity and/or expression. Alveolar monolayers were treated for 24 h with inhibitors of K(ATP), KvLQT1, and K(Ca) channels identified by PCR. Glibenclamide and clofilium (K(ATP) and KvLQT1 inhibitors) strongly reduced basal transepithelial current, amiloride-sensitive Na(+) current, and forskolin-activated Cl(-) currents, whereas pinacidil, a K(ATP) activator, increased them. Interestingly, K(+) inhibitors or membrane depolarization (induced by valinomycin in high-K(+) medium) decreased alpha-, beta-, and gamma-ENaC and CFTR mRNA. alpha-ENaC and CFTR proteins also declined after glibenclamide or clofilium treatment. Conversely, pinacidil augmented ENaC and CFTR mRNAs and proteins. Since alveolar fluid transport was found to be driven, at least in part, by Na(+) transport through ENaC, we tested the impact of K(+) channel modulators on fluid absorption across alveolar monolayers. We found that glibenclamide and clofilium reduced fluid absorption to a level similar to that seen in the presence of amiloride, whereas pinacidil slightly enhanced it. Long-term regulation of ENaC and CFTR expression by K(+) channel activity could benefit patients with pulmonary diseases affecting ion transport and fluid clearance.
authors.affiliation: Centre de recherche, Centre hospitalier de l'Université de Montréal-Hôtel-Dieu, Université de Montréal, Montréal, Québec, Canada.
authors.forename: Claudie
authors.initials: C
authors.lastname: Leroy
authors.forename: Anik
authors.initials: A
authors.lastname: Privé
authors.forename: Jean-Charles
authors.initials: JC
authors.lastname: Bourret
authors.forename: Yves
authors.initials: Y
authors.lastname: Berthiaume
authors.forename: Pasquale
authors.initials: P
authors.lastname: Ferraro
authors.forename: Emmanuelle
authors.initials: E
authors.lastname: Brochiero
mesh_terms: D000042:Absorption; D000818:Animals; D002478:Cells, Cultured; D019005:Cystic Fibrosis Transmembrane Conductance Regulator / Q000235:genetics*; D053503:Epithelial Sodium Channels / Q000235:genetics*; D054086:KATP Channels; D007700:Kinetics; D008297:Male; D016133:Polymerase Chain Reaction; D024661:Potassium Channels, Inwardly Rectifying / Q000502:physiology*; D011650:Pulmonary Alveoli / Q000502:physiology*; D012333:RNA, Messenger / Q000235:genetics; D051381:Rats; D017207:Rats, Sprague-Dawley; D020545:Respiratory Mucosa / Q000166:cytology / Q000502:physiology*
chemical_list: D053503:Epithelial Sodium Channels; D054086:KATP Channels; D024661:Potassium Channels, Inwardly Rectifying; D012333:RNA, Messenger; C092520:uK-ATP-1 potassium channel; D019005:Cystic Fibrosis Transmembrane Conductance Regulator