The objective of this application is to conduct studies to understand mechanisms by which hypoxia followed by reoxygenation (a model of ischemia/reperfusion) causes tissue injury. The applicant hypothesizes that reoxygenation increases the activity of a novel cell membrane oxidase that results in the enhanced production of deleterious reactive oxygen species. The proposed studies will use various endothelial cells in culture to investigate the role of the membrane associated oxidase activity as well as xanthine oxidase and nitric oxide synthase in the generation of reactive oxygen species under conditions of hypoxia and reoxygenation.
The specific aims are: 1) to investigate the presence of an NADPH oxidase in plasma membranes of endothelial cells by Northern and Western blot analysis; 2) to investigate the role of xanthine oxidase in the generation of intracellular reactive oxygen species and the mechanisms by which allopurinol and other antioxidants exert protective effects; and 3) to investigate the role of nitric oxide synthase in the pathogenesis of tissue injury by hypoxia and reoxygenation. During performance of the project, it is anticipated that Dr. Zulueta will become expert in facets of cellular biology and oxidant chemistry and will learn contemporary procedures for studies of cellular molecular biology and PCR technology.

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Minority School Faculty Development Awards (K14)
Project #
1K14HL003368-01
Application #
2211605
Study Section
Special Emphasis Panel (ZHL1-CCT-L (F1))
Project Start
1995-09-01
Project End
1998-08-31
Budget Start
1995-09-01
Budget End
1996-08-31
Support Year
1
Fiscal Year
1995
Total Cost
Indirect Cost
Name
Tufts University
Department
Type
DUNS #
City
Boston
State
MA
Country
United States
Zip Code
02111
Hassoun, P M; Yu, F S; Cote, C G et al. (1998) Upregulation of xanthine oxidase by lipopolysaccharide, interleukin-1, and hypoxia. Role in acute lung injury. Am J Respir Crit Care Med 158:299-305
Zulueta, J J; Sawhney, R; Yu, F S et al. (1997) Intracellular generation of reactive oxygen species in endothelial cells exposed to anoxia-reoxygenation. Am J Physiol 272:L897-902
Zulueta, J J; Bloom, S M; Rozansky, M I et al. (1996) Lung cancer in patients with bullous disease. Am J Respir Crit Care Med 154:519-22