Oxidative stress is involved the pathophysiology of a large number of diseases. This stress originates not only from normal aerobic metabolism, but also from the metabolism of foreign compounds, and as a direct result of the release of reactive oxygen species by certain cell types. Organisms have evolved antioxidant defenses against oxidative insults, which include antioxidant enzymes and through the consumption of antioxidant compounds. A very important enzyme involved in antioxidant defense is glutamate-cysteine ligase (GCL), the rate limiting enzyme for the synthesis of the cellular antioxidant glutathione (GSH). The primary goal of this project will be to investigate the role of GCL in defense against substances and conditions which induce oxidative damage to the liver. We propose to characterize a transgenic mouse model of GCL overexpression, and to assess the effects of modulating GCL expression on susceptibility to oxidant-induced damage to the liver. We propose to use three agents known to cause oxidative liver injury, namely acetaminophen, carbon tetrachioride, and tumor necrosis factor-alpha. Transgenic and wild-type (normal) littermates will be exposed to non-lethal doses of these agents, and sacrificed from 6 to 48 hours later. Liver tissue will be excised and examined for signs of oxidative damage, cellular necrosis and apoptosis, and biochemical and cellular measures of cell viability and function will be made. Such information will be useful in determining the functional significance of GCL in defense against reactive oxygen species and xenobiotics which induce oxidative stress, and lead to a better understanding of the significance of variable GCL expression in humans.

Agency
National Institute of Health (NIH)
Institute
National Institute of Environmental Health Sciences (NIEHS)
Type
Research Project (R01)
Project #
5R01ES010849-03
Application #
6785925
Study Section
Alcohol and Toxicology Subcommittee 4 (ALTX)
Program Officer
Maull, Elizabeth A
Project Start
2002-09-15
Project End
2006-07-31
Budget Start
2004-08-01
Budget End
2006-07-31
Support Year
3
Fiscal Year
2004
Total Cost
$260,757
Indirect Cost
Name
University of Washington
Department
Public Health & Prev Medicine
Type
Schools of Public Health
DUNS #
605799469
City
Seattle
State
WA
Country
United States
Zip Code
98195
Lim, Jinhwan; Nakamura, Brooke N; Mohar, Isaac et al. (2015) Glutamate Cysteine Ligase Modifier Subunit (Gclm) Null Mice Have Increased Ovarian Oxidative Stress and Accelerated Age-Related Ovarian Failure. Endocrinology 156:3329-43
Mohar, Isaac; Stamper, Brendan D; Rademacher, Peter M et al. (2014) Acetaminophen-induced liver damage in mice is associated with gender-specific adduction of peroxiredoxin-6. Redox Biol 2:377-87
Bir, Shyamal C; Shen, Xinggui; Kavanagh, Terrance J et al. (2013) Control of angiogenesis dictated by picomolar superoxide levels. Free Radic Biol Med 63:135-42
Lim, Jinhwan; Lawson, Gregory W; Nakamura, Brooke N et al. (2013) Glutathione-deficient mice have increased sensitivity to transplacental benzo[a]pyrene-induced premature ovarian failure and ovarian tumorigenesis. Cancer Res 73:908-17
Nakamura, Brooke N; Mohar, Isaac; Lawson, Gregory W et al. (2012) Increased sensitivity to testicular toxicity of transplacental benzo[a]pyrene exposure in male glutamate cysteine ligase modifier subunit knockout (Gclm-/-) mice. Toxicol Sci 126:227-41
Callegari, Andrea; Liu, Yuhua; White, Collin C et al. (2011) Gain and loss of function for glutathione synthesis: impact on advanced atherosclerosis in apolipoprotein E-deficient mice. Arterioscler Thromb Vasc Biol 31:2473-82
Nakamura, Brooke N; Fielder, Thomas J; Hoang, Yvonne D et al. (2011) Lack of maternal glutamate cysteine ligase modifier subunit (Gclm) decreases oocyte glutathione concentrations and disrupts preimplantation development in mice. Endocrinology 152:2806-15
McGuane, Jonathan T; Debrah, Julianna E; Sautina, Laura et al. (2011) Relaxin induces rapid dilation of rodent small renal and human subcutaneous arteries via PI3 kinase and nitric oxide. Endocrinology 152:2786-96
Krejsa, Cecile M; Franklin, Christopher C; White, Collin C et al. (2010) Rapid activation of glutamate cysteine ligase following oxidative stress. J Biol Chem 285:16116-24
Haque, Jamil A; McMahan, Ryan S; Campbell, Jean S et al. (2010) Attenuated progression of diet-induced steatohepatitis in glutathione-deficient mice. Lab Invest 90:1704-17

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