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Am J Physiol Lung Cell Mol Physiol 274: L369-L377, 1998;
1040-0605/98 $5.00
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Vol. 274, Issue 3, L369-L377, March 1998

Nitric oxide inhibits Na+ absorption across cultured alveolar type II monolayers

Yi Guo1, Michael D. Duvall1, John P. Crow1, and Sadis Matalon1,2,3

Departments of 1 Anesthesiology, 2 Physiology and Biophysics, and 3 Pediatrics, University of Alabama at Birmingham, Birmingham, Alabama 35294

We examined the mechanisms by which nitric oxide (· NO) decreased vectorial Na+ transport across confluent monolayers of rat alveolar type II (ATII) cells grown on permeable supports. Amiloride (10 µM) applied to the apical side of monolayers inhibited ~90% of the equivalent (Ieq) and the short-circuit (Isc) current, with an half-maximal inhibitory concentration (IC50) of 0.85 µM, indicating that Na+ entry into ATII cells occurred through amiloride-sensitive Na+ channels. · NO generated by spermine NONOate and papa NONOate added to both sides of the monolayers decreased Ieq and increased transepithelial resistance in a concentration-dependent fashion (IC50 = 0.4 µM · NO). These changes were prevented or reversed by addition of oxyhemoglobin (50 µM). Incubation of ATII monolayers with 8-bromoguanosine 3',5'-cyclic monophosphate (400 µM) had no effect on transepithelial Na+ transport. When the basolateral membranes of ATII cells were permeabilized with amphotericin B (10 µM) in the presence of a mucosal-to-serosal Na+ gradient (145:25 mM), · NO (generated by 100 µM papa NONOate) inhibited ~60% of the amiloride-sensitive Isc. In addition, after permeabilization of the apical membranes, · NO inhibited the Isc [a measure of Na+-K+-adenosinetriphosphatase (ATPase) activity] by ~60%. We concluded that · NO at noncytotoxic concentrations decreased Na+ absorption across cultured ATII monolayers by inhibiting both the amiloride-sensitive Na+ channels and Na+-K+-ATPase through guanosine 3',5'-cyclic monophosphate-independent mechanisms.

ion channels; amiloride; N-ethyl-N-isopropyl amiloride; short-circuit current; sodium ion-potassium ion-adenosinetriphosphatase; oxyhemoglobin; guanosine 3',5'-cyclic monophosphate; amphotericin B; active transport


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