Alveolar epithelial ion and fluid transport: Glucocorticoid-stimulated lung epithelial Na+ transport is associated with regulated ENaC and sgk1 expression

Abstract

H441 cells, a bronchiolar epithelial cell line, develop a glucocorticoid-regulated amiloride-sensitive Na+ transport pathway on permeable supports (R. Sayegh, S. D. Auerbach, X. Li, R. Loftus, R. Husted, J. B. Stokes, and C. P. Thomas. J Biol Chem 274: 12431-12437, 1999). To understand its molecular basis, we examined the effect of glucocorticoids (GC) on epithelial Na+ channel (ENaC)-α, -β, and -γ and sgk1 expression and determined the biophysical properties of Na+ channels in these cells. GC stimulated the expression of ENac-α, -β, and -γ and sgk1 mRNA, with the first effect seen by 1 h. These effects were abolished by actinomycin D, but not by cycloheximide, indicating a direct stimulatory effect on ENaC and sgk1 mRNA synthesis. The GC effect on transcription of ENaC-α mRNA was accompanied by a significant increase in ENaC-α protein levels. GC also stimulated ENaC-α, β, and -γ and sgk1 mRNA expression in A549 cells, an alveolar type II cell line. To determine the biophysical properties of the Na+ channel, single-channel currents were recorded from cell-attached H441 membranes. An Na+-selective channel with slow kinetics and a slope conductance of 10.8 pS was noted, properties similar to ENaC-α, β, and -γ expressed in Xenopus laevis oocytes. These experiments indicate that amiloride-sensitive Na+ transport is mediated through classic ENaC channels in human lung epithelia and that GC-regulated Na+ transport is accompanied by increased transcription of each of the component subunits and sgk1.