Influence of glutathione-S-transferase (GST) inhibition on lung epithelial cell injury: role of oxidative stress and metabolism

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Title: Influence of glutathione-S-transferase (GST) inhibition on lung epithelial cell injury: role of oxidative stress and metabolism
Authors: Fletcher, ME
Boshier, PR
Wakabayashi, K
Keun, HC
Smolenski, RT
Kirkham, PA
Adcock, IM
Barton, PJ
Takata, M
Marczin, N
Item Type: Journal Article
Abstract: Oxidant-mediated tissue injury is key to the pathogenesis of acute lung injury. Glutathione-S-transferases (GSTs) are important detoxifying enzymes that catalyze the conjugation of glutathione with toxic oxidant compounds and are associated with acute and chronic inflammatory lung diseases. We hypothesized that attenuation of cellular GST enzymes would augment intracellular oxidative and metabolic stress and induce lung cell injury. Treatment of murine lung epithelial cells with GST inhibitors, ethacrynic acid (EA), and caffeic acid compromised lung epithelial cell viability in a concentration-dependent manner. These inhibitors also potentiated cell injury induced by hydrogen peroxide (H2O2), tert-butyl-hydroperoxide, and hypoxia and reoxygenation (HR). SiRNA-mediated attenuation of GST-π but not GST-μ expression reduced cell viability and significantly enhanced stress (H2O2/HR)-induced injury. GST inhibitors also induced intracellular oxidative stress (measured by dihydrorhodamine 123 and dichlorofluorescein fluorescence), caused alterations in overall intracellular redox status (as evidenced by NAD+/NADH ratios), and increased protein carbonyl formation. Furthermore, the antioxidant N-acetylcysteine completely prevented EA-induced oxidative stress and cytotoxicity. Whereas EA had no effect on mitochondrial energetics, it significantly altered cellular metabolic profile. To explore the physiological impact of these cellular events, we used an ex vivo mouse-isolated perfused lung model. Supplementation of perfusate with EA markedly affected lung mechanics and significantly increased lung permeability. The results of our combined genetic, pharmacological, and metabolic studies on multiple platforms suggest the importance of GST enzymes, specifically GST-π, in the cellular and whole lung response to acute oxidative and metabolic stress. These may have important clinical implications.
Issue Date: 15-Jun-2015
Date of Acceptance: 1-Apr-2015
URI: http://hdl.handle.net/10044/1/62132
DOI: https://dx.doi.org/10.1152/ajplung.00220.2014
ISSN: 1040-0605
Publisher: American Physiological Society
Start Page: L1274
End Page: L1285
Journal / Book Title: AMERICAN JOURNAL OF PHYSIOLOGY-LUNG CELLULAR AND MOLECULAR PHYSIOLOGY
Volume: 308
Issue: 12
Copyright Statement: © 2015 the American Physiological Society
Sponsor/Funder: Wellcome Trust
Funder's Grant Number: 093080/Z/10/Z
Keywords: Science & Technology
Life Sciences & Biomedicine
Physiology
Respiratory System
N-acetylcysteine
ethacrynic acid
caffeic acid
reactive oxygen species
viability
PERFORMANCE LIQUID-CHROMATOGRAPHY
ETHACRYNIC-ACID
IN-VIVO
MYOCARDIAL-INFARCTION
PULMONARY-EDEMA
TRANSPLANTATION
GENOTYPE
MOUSE
OVEREXPRESSION
POLYMORPHISMS
N-acetylcysteine
caffeic acid
ethacrynic acid
reactive oxygen species
viability
Animals
Antioxidants
Blotting, Western
Caffeic Acids
Cell Membrane Permeability
Cell Survival
Cells, Cultured
Enzyme Inhibitors
Epithelial Cells
Ethacrynic Acid
Glutathione
Glutathione Transferase
Hydrogen Peroxide
Immunoenzyme Techniques
Lung Injury
Metabolomics
Mice
Oxidative Stress
RNA, Messenger
RNA, Small Interfering
Real-Time Polymerase Chain Reaction
Reverse Transcriptase Polymerase Chain Reaction
Cells, Cultured
Epithelial Cells
Animals
Mice
Hydrogen Peroxide
Caffeic Acids
Ethacrynic Acid
Glutathione Transferase
Glutathione
RNA, Small Interfering
RNA, Messenger
Enzyme Inhibitors
Antioxidants
Blotting, Western
Immunoenzyme Techniques
Reverse Transcriptase Polymerase Chain Reaction
Cell Membrane Permeability
Cell Survival
Oxidative Stress
Lung Injury
Metabolomics
Real-Time Polymerase Chain Reaction
Science & Technology
Life Sciences & Biomedicine
Physiology
Respiratory System
N-acetylcysteine
ethacrynic acid
caffeic acid
reactive oxygen species
viability
PERFORMANCE LIQUID-CHROMATOGRAPHY
ETHACRYNIC-ACID
IN-VIVO
MYOCARDIAL-INFARCTION
PULMONARY-EDEMA
TRANSPLANTATION
GENOTYPE
MOUSE
OVEREXPRESSION
POLYMORPHISMS
0606 Physiology
1116 Medical Physiology
Respiratory System
Publication Status: Published
Online Publication Date: 2015-06-15
Appears in Collections:Division of Surgery
National Heart and Lung Institute
Airway Disease
Division of Cancer



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