This project proposes to determine factors that regulate glutathione synthesis in the lung and lung cells under normoxic conditions and conditions of oxidant exposure. We have obtained preliminary data which suggest that the level of GSH in pulmonary artery endothelial cells in culture is correlated with the rate of uptake (transport) of glutamic acid and cystine. Our first studies will include further characterization of this amino acid transport system and determination of the relationship between GSH levels and endothelial cell uptake of cystine and glutamate during different exposures to oxidant stresses, growth conditions and phases of the cell cycle. We will then investigate other lung cells including fibroblasts, smooth muscle cells, macrophages, and alveolar type II epithelial cells to see if GSH levels in these cells are also controlled by availability of amino acid precursors, or if other mechanisms of control such as direct uptake of intact GSH, transport of gamma-glutamyldipeptides or regulation of the feedback inhibition of glutathione are predominant control factors in these cells. We have succeeded in increasing GSH levels up to five-fold in cultured endothelial cells by treatment with low concentrations of diethylmaleate (DEM). We will attempt to produce similar increases in GSH in vivo by low doses of DEM over a 24 hr period. We will also use DEM and other treatments to increase cell levels of GSH and determine which types of oxidant stresses can be protected against by increases in GSH levels and during what time period GSH availability is most critical. Finally, we will attempt to determine whether blood or erythrocyte GSH levels reflect either the tissue GSH status or the general state of oxidant stress first in rats and then in patients undergoing oxygen therapy for treatments of disease conditions. The results of these studies should provide new insights into the development of strategies to protect against oxidant injury.

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Research Project (R01)
Project #
5R01HL032824-07
Application #
3344329
Study Section
Respiratory and Applied Physiology Study Section (RAP)
Project Start
1989-09-01
Project End
1993-03-31
Budget Start
1990-04-01
Budget End
1991-03-31
Support Year
7
Fiscal Year
1990
Total Cost
Indirect Cost
Name
University of Texas Health Science Center San Antonio
Department
Type
Schools of Medicine
DUNS #
800772162
City
San Antonio
State
TX
Country
United States
Zip Code
78229
Deneke, S M; Harford, P H; Lee, K Y et al. (1997) Induction of cystine transport and other stress proteins by disulfiram: effects on glutathione levels in cultured cells. Am J Respir Cell Mol Biol 17:227-34
Deneke, S M; Susanto, I; Vogel, K A et al. (1995) Mechanisms of use of extracellular glutathione by lung epithelial cells and pulmonary artery endothelial cells. Am J Respir Cell Mol Biol 12:662-8
Bukowski, D M; Deneke, S M; Lawrence, R A et al. (1995) A noninducible cystine transport system in rat alveolar type II cells. Am J Physiol 268:L21-6
Morales, C F; Anzueto, A; Andrade, F et al. (1994) Buthionine sulfoximine treatment impairs rat diaphragm function. Am J Respir Crit Care Med 149:915-9
Jenkinson, S G; Lawrence, R A; Zamora, C A et al. (1994) Induction of intracellular glutathione in alveolar type II pneumocytes following BCNU exposure. Am J Physiol 266:L125-30
Levine, S M; Anzueto, A; Peters, J I et al. (1994) Single lung transplantation in patients with systemic disease. Chest 105:837-41
Anzueto, A; Brassard, J M; Andrade, F H et al. (1994) Effects of hyperoxia on rat diaphragm function. J Appl Physiol 77:63-8
Morales, C F; Anzueto, A; Andrade, F et al. (1993) Diethylmaleate produces diaphragmatic impairment after resistive breathing. J Appl Physiol 75:2406-11
Deneke, S M; Lawrence, R A; Jenkinson, S G (1992) Endothelial cell cystine uptake and glutathione increase with N,N-bis(2-chloroethyl)-N-nitrosourea exposure. Am J Physiol 262:L301-4
Deneke, S M (1992) Induction of cystine transport in bovine pulmonary artery endothelial cells by sodium arsenite. Biochim Biophys Acta 1109:127-31

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