The overall goal of this proposal is to study the mechanisms by which partially reduced oxygen species mediate lung cell injury. Aortic and pulmonary artery endothelial cells and type II aveolar epithelial cells will be used as in vitro models because a data base exists for both the in vitro and in vivo responses of these cell types to oxidant stress. To accomplish this, proposed studies are designed to illuminate biochemical mechanisms of oxidant lung injury. Thus, the following specific aims will be addressed: 1. Establish an endothelial cell and type II aveolar epithelial cell culture models for the study of free radical mediated lung injury. 2. Study cytoplasmically, membrane and extracellularly generated free radicals using EPR techniques. 3. Cellular formation of free radicals will be monitored and modulated using hyperoxia and uncouplers of electron transport. 4. Cellular metabolism of free radicals will be studied using free radical scavengers and by liposome mediated augmentation of intracellular antioxidant defenses. 5. Cellular indices of free radical injury and modulation of injury will be studied using metabolic indicators and EPR measurement of structural changes in membrane microdomains. This research proposal represents a combination of lung cell biology and sophisticated spectroscopic approaches towards the identification of specific free radicals and their affects on membrane structure. Pharmacological manipulation of antioxidant defenses using liposomes containing superoxide dismutase or catalase will be potentially beneficial in many metabolic situations. Finally, completion of the above specific aims will provide insight into the etiology of oxidant lung injury.

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Research Project (R01)
Project #
1R01HL033550-01A1
Application #
3345524
Study Section
Pathology A Study Section (PTHA)
Project Start
1985-08-01
Project End
1988-07-31
Budget Start
1985-08-01
Budget End
1986-07-31
Support Year
1
Fiscal Year
1985
Total Cost
Indirect Cost
Name
Duke University
Department
Type
Schools of Medicine
DUNS #
071723621
City
Durham
State
NC
Country
United States
Zip Code
27705
Zhang, B; Centra, M; Cao, G L et al. (1996) Penicillin G-induced microbicidal activity of endothelial cells cultured on gelfoam blocks. J Infect Dis 174:1001-9
Halpern, H J; Peric, M; Yu, C et al. (1996) In vivo spin-label murine pharmacodynamics using low-frequency electron paramagnetic resonance imaging. Biophys J 71:403-9
Pou, S; Bhan, A; Bhadti, V S et al. (1995) The use of fluorophore-containing spin traps as potential probes to localize free radicals in cells with fluorescence imaging methods. FASEB J 9:1085-90
Rosen, G M; Pou, S; Ramos, C L et al. (1995) Free radicals and phagocytic cells. FASEB J 9:200-9
Ramos, C L; Pou, S; Rosen, G M (1995) Effect of anti-inflammatory drugs on myeloperoxidase-dependent hydroxyl radical generation by human neutrophils. Biochem Pharmacol 49:1079-84
Pou, S; Nguyen, S Y; Gladwell, T et al. (1995) Does peroxynitrite generate hydroxyl radical? Biochim Biophys Acta 1244:62-8
Halpern, H J; Yu, C; Barth, E et al. (1995) In situ detection, by spin trapping, of hydroxyl radical markers produced from ionizing radiation in the tumor of a living mouse. Proc Natl Acad Sci U S A 92:796-800
Pou, S; Keaton, L; Surichamorn, W et al. (1994) Can nitric oxide be spin trapped by nitrone and nitroso compounds? Biochim Biophys Acta 1201:118-24
Pou, S; Ramos, C L; Gladwell, T et al. (1994) A kinetic approach to the selection of a sensitive spin trapping system for the detection of hydroxyl radical. Anal Biochem 217:76-83
Rosen, G M; Pou, S; Britigan, B E et al. (1994) Spin trapping of hydroxyl radicals in biological systems. Methods Enzymol 233:105-11

Showing the most recent 10 out of 37 publications