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Am. J. Respir. Cell Mol. Biol., Volume 25, Number 5, November, 2001 554-561

Hypoxia Reduces Alveolar Epithelial Sodium and Fluid Transport in Rats
Reversal by beta -Adrenergic Agonist Treatment

Marie Laure Vivona, Michael Matthay, Marcel Blot Chabaud, Gérard Friedlander, and Christine Clerici

Department of Physiology, Faculté de Médecine Léonard de Vinci, Université Paris 1 3, Paris; Cardiovascular Research Institute, University of California, San Francisco, San Francisco, California; Unité INSERM 478, Faculté de Médecine Xavier Bichat, Université Paris 7, Paris; and Unité INSERM 426 Faculté de Médecine de Bichat, Université Paris 7, Paris, France

In cultured alveolar epithelial cells, hypoxia induces a downregulation of the two main Na proteins, the epithelial Na channel (ENaC) and the Na,K-ATPase. However, the in vivo effects of hypoxia on alveolar epithelial transport have not been well studied. Therefore, the objectives of this study were to investigate in an in vivo rat model if hypoxia induces a reduction in vectorial Na and fluid transport across the alveolar epithelium in vivo, and if a change in net fluid transport is associated with modification in the expression and/or activity of Na transport proteins. Rats were exposed to 8% O2 from 3 to 24 h. Hypoxia induced a progressive decrease in alveolar liquid clearance (ALC) reaching 50% at 24 h, an effect that was related primarily to a decrease in amiloride-sensitive transepithelial Na transport. On RNase protection assay of alveolar type II (ATII) cells isolated immediately after hypoxic exposure, steady state levels of mRNA were increased for alpha -rENaC and beta 1-Na, K-ATPase, whereas the levels of gamma -rENaC and alpha 1-Na,K-ATPase were unchanged. On Western blots of ATII cell membranes, alpha -ENaC subunit protein slightly increased, whereas the amount of alpha 1- and beta 1-Na,K-ATPase protein were unchanged with hypoxia. Thus, the decrease in transepithelial Na transport was not explained by a parallel change in gene expression or the quantity of transport proteins. Interestingly, hypoxia-induced decrease in ALC was completely reversed by intra-alveolar administration of the beta 2 agonist, terbutaline (10-4 M). These results suggest that hypoxia-induced decrease in Na transport is not simply related to a downregulation of Na transport proteins but rather to a decrease in Na protein activity by either internalization of the proteins and/or direct alteration of the protein in the membrane. The dramatic increase of ALC with beta 2-agonist therapy indicates that the decrease of transepithelial Na and fluid transport during hypoxia is rapidly reversible, a finding of major clinical significance.

Abbreviations: alveolar liquid clearance, ALC; alveolar type II, ATII; ethylenediaminetetraacetic acid, EDTA; messenger RNA, mRNA.




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