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Am J Physiol Lung Cell Mol Physiol 275: L764-L770, 1998;
1040-0605/98 $5.00
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Vol. 275, Issue 4, L764-L770, October 1998

O2-induced ENaC expression is associated with NF-kappa B activation and blocked by superoxide scavenger

Bijan Rafii1,2, A. Keith Tanswell1,3, Gail Otulakowski1,2, Olli Pitkänen4, Rose Belcastro-Taylor1,3, and Hugh O'Brodovich1,2

1 Medical Research Council Group in Lung Development and Divisions of 2 Respiratory and 3 Neonatal Research, Hospital for Sick Children Research Institute and Department of Paediatrics, University of Toronto, Toronto, Ontario, Canada M5G 1X8; and 4 Hospital for Children and Adolescents, University of Helsinki, 00290 Helsinki, Finland

Cultured rat fetal distal lung epithelial cells (FDLEs), when switched from fetal (3%) to postnatal (21%) O2 concentrations, have increased epithelial Na+ channel (ENaC) mRNA levels and amiloride-sensitive Na+ transport [O. Pitkänen, A. K. Tanswell, G. Downey, and H. O'Brodovich. Am. J. Physiol. 270 (Lung Cell. Mol. Physiol. 14): L1060-L1066, 1996]. The mechanisms by which O2 mediates these effects are unknown. After isolation, FDLEs were kept at 3% O2 overnight, then switched to 21% O2 (3-21% O2 group) or maintained at 3% O2 (3-3% O2 group) for 48 h. The amiloride-sensitive short-circuit current (Isc) in the 3-21% O2 group was double that in the 3-3% O2 group. Amiloride-sensitive Isc could not be induced by medium conditioned by 21% O2-exposed FDLEs but was reversed by returning the cells to 3% O2. Neither the cyclooxygenase inhibitor ibuprofen, liposome-encapsulated catalase, nor hydroperoxide scavengers (U-74389G or Trolox) blocked the O2-induced amiloride-sensitive Isc. In contrast, the cell-permeable superoxide scavenger tetramethylpiperidine-N-oxyl (TEMPO) eliminated the O2-induced increases in amiloride-sensitive Isc and ENaC mRNA levels. The switch from 3 to 21% O2 induced the transcription factor nuclear factor-kappa B, which could also be blocked by TEMPO. We conclude that 1) the O2-induced increase in amiloride-sensitive Isc is reversible and 2) the O2-induced increase in amiloride-sensitive Isc and ENaC mRNA levels is associated with activation of nuclear factor-kappa B and may be mediated, at least in part, by superoxide.

epithelial sodium channel; nuclear factor-kappa B; amiloride; type II epithelium; tetramethylpiperidine-N-oxyl; reactive oxygen species


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