Hypoxia and beta 2-agonists regulate cell surface expression of the epithelial sodium channel in native alveolar epithelial cells.
Alveolar hypoxia may impair sodium-dependent alveolar fluid transport and induce pulmonary edema in rat and human lung, an effect that can be prevented by the inhalation of beta(2)-agonists. To investigate the mechanism of beta(2)-agonist-mediated stimulation of sodium transport under conditions of moderate hypoxia, we examined the effect of terbutaline on epithelial sodium channel (ENaC) expression and activity in cultured rat alveolar epithelial type II cells exposed to 3% O(2) for 24 h. Hypoxia reduced transepithelial sodium current and amiloride-sensitive sodium channel activity without decreasing ENaC subunit mRNA or protein levels. The functional decrease was associated with reduced abundance of ENaC subunits (especially beta and gamma) in the apical membrane of hypoxic cells, as quantified by biotinylation. cAMP stimulation with terbutaline reversed the hypoxia-induced decrease in transepithelial sodium transport by stimulating sodium channel activity and markedly increased the abundance of beta-and gamma-ENaC in the plasma membrane of hypoxic cells. The effect of terbutaline was prevented by brefeldin A, a blocker of anterograde transport. These novel results establish that hypoxia-induced inhibition of amiloride-sensitive sodium channel activity is mediated by decreased apical expression of ENaC subunits and that beta(2)-agonists reverse this effect by enhancing the insertion of ENaC subunits into the membrane of hypoxic alveolar epithelial cells.
Planès
Carole
C
Department of Physiology, INSERM U426, Université Paris 7, 75018 Paris, France.
Blot-Chabaud
Marcel
M
Matthay
Michael A
MA
Couette
Sylviane
S
Uchida
Tokujiro
T
Clerici
Christine
C
D000318:Adrenergic beta-Agonists / Q000494:pharmacology*; D000818:Animals; D001692:Biological Transport; D019921:Biotinylation; D015153:Blotting, Western; D020126:Brefeldin A / Q000494:pharmacology; D002462:Cell Membrane / Q000378:metabolism*; D000242:Cyclic AMP / Q000378:metabolism; D004594:Electrophysiology; D004847:Epithelial Cells / Q000378:metabolism*; D053503:Epithelial Sodium Channels; D000860:Hypoxia; D008297:Male; D010100:Oxygen / Q000378:metabolism; D011650:Pulmonary Alveoli / Q000166:cytology*; D012333:RNA, Messenger / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D012964:Sodium / Q000378:metabolism; D015222:Sodium Channels / Q000378:metabolism*; D013726:Terbutaline / Q000494:pharmacology; D013997:Time Factors
D000318:Adrenergic beta-Agonists; D053503:Epithelial Sodium Channels; D012333:RNA, Messenger; D015222:Sodium Channels; D020126:Brefeldin A; D012964:Sodium; D000242:Cyclic AMP; D013726:Terbutaline; D010100:Oxygen