Nitric oxide inhibits lung sodium transport through a cGMP-mediated inhibition of epithelial cation channels.
We used the patch-clamp technique to study the effect of nitric oxide (NO) on a cation channel in rat type II pneumocytes [alveolar type II (AT II) cells]. Single-channel recordings from the apical surface of AT II cells in primary culture showed a predominant cation channel with a conductance of 20.6 +/- 1.1 (SE) pS (n = 9 cell-attached patches) and Na(+)-to-K+ selectivity of 0.97 +/- 0.07 (n = 7 cell-attached patches). An NO donor, S-nitrosoglutathione (GSNO; 100 microM), inhibited the basal cation-channel activity by 43% [open probability (Po), control 0.28 +/- 0.05 vs. GSNO 0.16 +/- 0.03; P < 0.001; n = 16 cell-attached patches], with no significant change in the conductance. GSNO reduced the Po by reducing channel mean open and increasing mean closed times. GSNO inhibition was reversed by washout. The inhibitory effect of NO was confirmed by using a second donor of NO, S-nitroso-N-acetylpenicillamine (100 microM; Po, control 0.53 +/- 0.05 vs. S-nitroso-N-acetylpenicillamine 0.31 +/- 0.04; -42%; P < 0.05; n = 5 cell-attached patches). The GSNO effect was blocked by methylene blue (a blocker of guanylyl cyclase; 100 microM), suggesting a role for cGMP. The permeable analog of cGMP, 8-bromo-cGMP (8-BrcGMP; 1 mM), inhibited the cation channel in a manner similar to GSNO (Po, control 0.38 +/- 0.06 vs. 8-BrcGMP 0.09 +/- 0.02; P < 0.05; n = 7 cell-attached patches). Pretreatment of cells with 1 microM KT-5823 (a blocker of protein kinase G) abolished the inhibitory effect of GSNO. The NO inhibition of channels was not due to changes in cell viability. Intracellular cGMP was found to be elevated in AT II cells treated with NO (control 13.4 +/- 3.6 vs. GSNO 25.4 +/- 4.1 fmol/ml; P < 0.05; n = 6 cell-attached patches). We conclude that NO suppresses the activity of an Na(+)-permeant cation channel on the apical surface of AT II cells. This action appears to be mediated by a cGMP-dependent protein kinase.
Jain
L
L
Department of Pediatrics, Emory University School of Medicine, Atlanta, Georgia 30322, USA.
Chen
X J
XJ
Brown
L A
LA
Eaton
D C
DC
D000818:Animals; D001692:Biological Transport / Q000187:drug effects; D002412:Cations / Q000378:metabolism*; D006152:Cyclic GMP / Q000378:metabolism / Q000502:physiology*; D004848:Epithelium / Q000378:metabolism; D007473:Ion Channels / Q000037:antagonists & inhibitors* / Q000378:metabolism; D008168:Lung / Q000378:metabolism*; D009569:Nitric Oxide / Q000494:pharmacology*; D011494:Protein Kinases / Q000502:physiology; D051381:Rats; D017207:Rats, Sprague-Dawley; D012964:Sodium / Q000378:metabolism*; D013997:Time Factors
D002412:Cations; D007473:Ion Channels; D009569:Nitric Oxide; D012964:Sodium; D011494:Protein Kinases; D006152:Cyclic GMP