Protection of lungs from hyperoxic injury: gene expression analysis of cyclosporin A therapy.
We have previously shown that cyclosporin A (CsA), an inhibitor of protein phosphatase 2B (calcineurin), attenuates hyperoxia-induced reductions in murine lung compliance. CsA protected against hyperoxia-induced changes in neutrophil infiltration, capillary congestion, edema, and hyaline membrane formation. Gene expression studies were conducted to identify the gene expression patterns underlying the protective effects of CsA during hyperoxic lung injury. After 72 h of simultaneous treatment with >95% oxygen and CsA (50 mg x kg(-1) x day(-1)), RNA was isolated from murine lungs. RNA from treated and untreated lungs was reverse transcribed to cDNA, competitively hybridized, and used to probe 8,734 complimentary DNAs on the Incyte mouse GEM 1 array. Several known genes and expressed sequence tags (ESTs) showed increased (GenBank accession numbers: AA125385, AA241295, W87197, syntaxin, and cyclin G) or decreased [AA036517, AA267567, AA217009, W82577, uteroglobin, stromal cell-derived factor 1, and surfactant protein C (SP-C)] expression after hyperoxia. Hyperoxia-stimulated reductions in SP-C gene expression were confirmed through Northern blot analysis. The increase in gene expression of one expressed sequence tag (AA125385) with hyperoxia was reversed by CsA treatment. Sequence data demonstrated that this EST has high homology to murine cyclin B1. Western blot analysis did not demonstrate any changes in distal lung cyclin B1 expression after hyperoxia. Protein expression of cyclin B1 in the distal lung was observed in the endothelial cells, bronchiolar epithelial cells, and both the type I and type II alveolar epithelial cells. Further analysis of cyclin B1 may elucidate the protective actions of CsA in hyperoxic injury.
Matthew
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Department of Molecular and Cellular Physiology, University of Cincinnati Medical Center, Cincinnati, Ohio 45267-0576, USA. mattheeh@email.uc.edu
Kutcher
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Dedman
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D000818:Animals; D019703:Calcineurin / Q000235:genetics; D019926:Cyclin B / Q000235:genetics; D056744:Cyclin B1; D016572:Cyclosporine / Q000494:pharmacology / Q000627:therapeutic use*; D015536:Down-Regulation / Q000187:drug effects / Q000235:genetics; D005260:Female; D020869:Gene Expression Profiling / Q000379:methods; D005786:Gene Expression Regulation / Q000187:drug effects*; D005796:Genes / Q000187:drug effects / Q000235:genetics; D018496:Hyperoxia / Q000188:drug therapy / Q000235:genetics* / Q000517:prevention & control*; D008171:Lung Diseases / Q000188:drug therapy / Q000235:genetics* / Q000517:prevention & control*; D051379:Mice; D008815:Mice, Inbred Strains; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods; D012313:RNA / Q000096:biosynthesis; D015854:Up-Regulation / Q000187:drug effects / Q000235:genetics
C534700:Ccnb1 protein, mouse; D019926:Cyclin B; D056744:Cyclin B1; D012313:RNA; D016572:Cyclosporine; D019703:Calcineurin