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Articles in PresS, published online ahead of print August 9, 2002
Am J Physiol Lung Cell Mol Physiol, 10.1152/ajplung.00464.2001
Submitted on December 4, 2001
Accepted on July 4, 2002
1 Department of Physiology, Faculte de Medecine Leonard de Vinci, Universite Paris 13 and INSERM U 426, Faculte Xavier Bichat, Bobigny, 93012, France
2 Division of Allergy, Pulmonary and Critical Care Medicine, Vanderbilt University School of Medicine, Nasville, USA
3 Division of Pulmonary and Critical Care Medicine, Weill Medical College, NY, USA
4 Cardiovascular Research Institute, University of California, San Francisco, California, USA
* To whom correspondence should be addressed. E-mail: christine.clerici{at}lmr.ap-hop-paris.fr.
We investigated the regulation of VEGF expression by hypoxia in cultured and freshly isolated rat alveolar epithelial cells (AEC). In vitro, hypoxia increased VEGF mRNA and protein levels with a maximal stimulation at 0% O2 for 18 hours. A similar upregulation VEGF of expression was found in freshly ATII cells isolated from rats exposed to 8% O2 for 24 hours. In vitro, the hypoxia-induced upregulation of VEGF mRNA was due to an increase in transcription rather than to an increase in RNA stability, since the half-life of VEGF mRNA was unchanged. The upregulation of VEGF mRNA by hypoxia was mimicked by cobalt chloride and desferrioxamine in normoxic AEC, and was not prevented by inhibitors of reactive oxygen species (ROS) suggesting that hypoxic VEGF regulation involved an O2 dependent protein that required ferrous ions but is independent of ROS generation. In polarized ATII cells, VEGF protein was secreted both at the apical and the basolateral sides. Similarly, in rats, VEGF was secreted in the broncho-alveolar lavage fluid. Hypoxia induced a 2-fold increase in VEGF protein at the apical side of ATII cells in culture and in BAL fluid. These findings suggest that release of VEGF synthetized by alveolar epithelial cells may not only target the endothelial cells, but also other alveolar cells, including macrophages and epithelial cells.
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