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1 Division of Oncology, Center for Applied Medical Research (CIMA), Pamplona, Navarra, Spain; Department of Histology and Pathology, University of Navarra School of Medicine, Pamplona, Navarra, Spain
2 Pulmonary Service, Clinica Universitaria; University of Navarro School of Medicine, Pamplona, Navarra, Spain
* To whom correspondence should be addressed. E-mail: jagorreta{at}unav.es.
Adrenomedullin (ADM) is upregulated independently by hypoxia and LPS, two key factors in the pathogenesis of ALI. This study evaluates the expression of ADM in ALI using experimental models combining both stimuli: an in vivo model of rats treated with LPS and acute normobaric hypoxia (9% O2), and an in vitro model of rat lung cell lines cultured with LPS and exposed to hypoxia (1% O2). ADM expression was analyzed by in situ hybridization, Northern blot, Western blot and RIA analyses. In the rat lung, combination of hypoxia and LPS treatments overcomes ADM induction occurring after each treatment alone. Using in situ techniques, the synergistic effect of both stimuli mainly correlates with ADM expression in inflammatory cells within blood vessels, and to a lesser extent to cells in the lung parenchyma and bronchiolar epithelial cells. In the in vitro model, hypoxia and hypoxia + LPS treatments caused a similar strong induction of ADM expression and secretion in epithelial and endothelial cell lines. In alveolar macrophages, however, LPSinduced ADM expression and secretion was further increased by the concomitant exposure to hypoxia, thus paralleling the in vivo response. In conclusion, ADM expression is highly induced in a variety of key lung cell types in this rat model of ALI by combination of hypoxia and LPS, suggesting an essential role for this mediator in this syndrome.
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