Modification of biophysical properties of lung epithelial Na(+) channels by dexamethasone.
There is considerable interest in identifying the basic mechanisms by which dexamethasone alters ion transport across the adult alveolar epithelium. Herein, we incubated synchronized A549 cells, a human alveolar epithelial cell line, with dexamethasone (1 microM) for 24-48 h. When normalized to HPRT (a housekeeping gene), A549 beta- and gamma-subunit mRNA levels for the human amiloride-sensitive epithelial sodium channel (hENaC), assessed by RT-PCR, increased by 1.6- and 17-fold respectively, compared with control values (P < 0.05). These changes were abolished by actinomycin D, indicating transcriptional regulation. Western blotting studies revealed that dexamethasone also increased expression of beta- and gamma-hENaC protein levels. In contrast, alpha-hENaC mRNA increased by onefold (P > 0.05) and alpha-hENaC protein level was unchanged. Incubation of A549 cells with dexamethasone increased their whole cell amiloride-sensitive sodium currents twofold and decreased the K(0.5) for amiloride from 833 +/- 69 to 22 +/- 5.4 nM (mean +/- SE; P < 0.01). Single channel recordings in the cell-attached mode showed that dexamethasone treatment increased single channel open time and open probability threefold and decreased channel conductance from 8.63 +/- 0.036 to 4. 4 +/- 0.027 pS (mean +/- SE; P < 0.01). We concluded that dexamethasone modulates the amiloride-sensitive Na(+) channels by differentially regulating the expression of beta- and gamma-subunits at the mRNA and protein levels in the human A549 cell line, with little effect on alpha-hENaC subunit.
Lazrak
A
A
Department of Anesthesiology, University of Alabama at Birmingham, Birmingham, Alabama 35233, USA.
Samanta
A
A
Venetsanou
K
K
Barbry
P
P
Matalon
S
S
D000287:Administration, Topical; D000893:Anti-Inflammatory Agents / Q000494:pharmacology*; D002460:Cell Line; D003907:Dexamethasone / Q000494:pharmacology*; D004553:Electric Conductivity; D004594:Electrophysiology; D053503:Epithelial Sodium Channels; D005938:Glucocorticoids; D006801:Humans; D008168:Lung / Q000166:cytology / Q000378:metabolism* / Q000502:physiology; D012333:RNA, Messenger / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015222:Sodium Channels / Q000187:drug effects* / Q000235:genetics / Q000502:physiology*
D000893:Anti-Inflammatory Agents; D053503:Epithelial Sodium Channels; D005938:Glucocorticoids; D012333:RNA, Messenger; D015222:Sodium Channels; D003907:Dexamethasone