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Am J Physiol Lung Cell Mol Physiol 277: L589-L595, 1999;
1040-0605/99 $5.00
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Vol. 277, Issue 3, L589-L595, September 1999

Mitogen-activated protein kinase pathway mediates hyperoxia-induced apoptosis in cultured macrophage cells

Irina Petrache1, Mary E. Choi2,3, Leo E. Otterbein4, Beek Yoke Chin4, Lin L. Mantell5, Stuart Horowitz5, and Augustine M. K. Choi2,6

6 Section of Pulmonary and Critical Care Medicine and 3 Section of Nephrology, Department of Internal Medicine, Yale University School of Medicine, New Haven 06520; 2 Connecticut Veterans Affairs HealthCare System, West Haven, Connecticut 06516; 5 Departments of Thoracic Cardiovascular Surgery and Pediatrics, The CardioPulmonary Research Institute, Winthrop-University Hospital, State University of New York at Stony Brook School of Medicine, Mineola, New York 11501; and 1 Division of Pulmonary and Critical Care Medicine and 4 Department of Environmental Health Sciences, The Johns Hopkins Medical Institutions, Baltimore, Maryland 21205

We have previously demonstrated that the lungs of mice can exhibit increased programmed cell death or apoptosis after hyperoxic exposure in vivo. In this report, we show that hyperoxic exposure in vitro can also induce apoptosis in cultured murine macrophage cells (RAW 264.7) as assessed by DNA-laddering, terminal deoxynucleotidyltransferase dUTP nick end-labeling, and nucleosomal assays. To further delineate the signaling pathway of hyperoxia-induced apoptosis in RAW 264.7 macrophages, we first show that hyperoxia can activate the mitogen-activated protein kinase (MAPK) pathway, the extracellular signal-regulated kinases (ERKs) p42/p44, in a time-dependent manner as assessed by increased phosphorylation of ERK1/ERK2 by Western blot analyses. Neither the c-Jun NH2-terminal kinase/stress-activated protein kinase nor the p38 MAPK was activated by hyperoxia in these cells. Chemical or genetic inhibition of the ERK p42/p44 MAPK pathway by PD-98059, a selective inhibitor of MAPK kinase, and dominant negative mutants of ERK, respectively, attenuated hyperoxia-induced apoptosis as assessed by DNA laddering and nucleosomal ELISAs. Taken together, our data suggest that hyperoxia can induce apoptosis in cultured murine macrophages and that the MAPK pathway mediates hyperoxia-induced apoptosis.

programmed cell death; oxygen; signal transduction; extracellular signal-regulated kinase; cell death


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