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B activation and blocked by superoxide
scavenger
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-
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-B and may be
mediated, at least in part, by superoxide.
epithelial sodium channel; nuclear factor-B; amiloride; type II
epithelium; tetramethylpiperidine-N-oxyl; reactive
oxygen species
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