skip to main content
OSTI.GOV title logo U.S. Department of Energy
Office of Scientific and Technical Information

Title: Transcription factor Nrf2 mediates an adaptive response to sulforaphane that protects fibroblasts in vitro against the cytotoxic effects of electrophiles, peroxides and redox-cycling agents

Abstract

Sulforaphane can stimulate cellular adaptation to redox stressors through transcription factor Nrf2. Using mouse embryonic fibroblasts (MEFs) as a model, we show herein that the normal homeostatic level of glutathione in Nrf2{sup -/-} MEFs was only 20% of that in their wild-type counterparts. Furthermore, the rate of glutathione synthesis following its acute depletion upon treatment with 3 {mu}mol/l sulforaphane was very substantially lower in Nrf2{sup -/-} MEFs than in wild-type cells, and the rebound leading to a {approx} 1.9-fold increase in glutathione that occurred 12-24 h after Nrf2{sup +/+} MEFs were treated with sulforaphane was not observed in Nrf2{sup -/-} fibroblasts. Wild-type MEFs that had been pre-treated for 24 h with 3 {mu}mol/l sulforaphane exhibited between 1.4- and 3.2-fold resistance against thiol-reactive electrophiles, including isothiocyanates, {alpha},{beta}-unsaturated carbonyl compounds (e.g. acrolein), aryl halides and alkene epoxides. Pre-treatment of Nrf2{sup +/+} MEFs with sulforaphane also protected against hydroperoxides (e.g. cumene hydroperoxide, CuOOH), free radical-generating compounds (e.g. menadione), and genotoxic electrophiles (e.g. chlorambucil). By contrast, Nrf2{sup -/-} MEFs were typically {approx} 50% less tolerant of these agents than wild-type fibroblasts, and sulforaphane pre-treatment did not protect the mutant cells against xenobiotics. To test whether Nrf2-mediated up-regulation of glutathione represents the major cytoprotective mechanismmore » stimulated by sulforaphane, 5 {mu}mol/l buthionine sulfoximine (BSO) was used to inhibit glutathione synthesis. In Nrf2{sup +/+} MEFs pre-treated with sulforaphane, BSO diminished intrinsic resistance and abolished inducible resistance to acrolein, CuOOH and chlorambucil, but not menadione. Thus Nrf2-dependent up-regulation of GSH is the principal mechanism by which sulforaphane pre-treatment induced resistance to acrolein, CuOOH and chlorambucil, but not menadione.« less

Authors:
;  [1];  [2];  [3];  [3]
  1. Biomedical Research Institute, Ninewells Hospital and Medical School, University of Dundee, Dundee DD1 9SY, Scotland (United Kingdom)
  2. Department of Pharmacy and Pharmacology, University of Bath, Claverton Down, Bath BA2 7AY (United Kingdom)
  3. Center for Tsukuba Advanced Research Alliance, University of Tsukuba, Tsukuba 305-8577 (Japan)
Publication Date:
OSTI Identifier:
21272577
Resource Type:
Journal Article
Journal Name:
Toxicology and Applied Pharmacology
Additional Journal Information:
Journal Volume: 237; Journal Issue: 3; Other Information: DOI: 10.1016/j.taap.2009.03.005; PII: S0041-008X(09)00115-X; Copyright (c) 2009 Elsevier Science B.V., Amsterdam, The Netherlands, All rights reserved; Country of input: International Atomic Energy Agency (IAEA); Journal ID: ISSN 0041-008X
Country of Publication:
United States
Language:
English
Subject:
60 APPLIED LIFE SCIENCES; ACROLEIN; BENZOQUINONES; CHLORAMBUCIL; CUMENE; CYSTEINE; EPOXIDES; FIBROBLASTS; GLUTATHIONE; ISOTHIOCYANATES; LIGASES; NEOPLASMS; PEROXIDES; TRANSCRIPTION FACTORS; TRANSFERASES

Citation Formats

Higgins, Larry G, Kelleher, Michael O, Eggleston, Ian M, Itoh, Ken, Yamamoto, Masayuki, Department of Medical Biochemistry, Tohoku University Graduate School of Medicine, Sendai 980-8577, ERATO Environmental Response Project, Japan Science and Technology Corporation, Tsukuba 305-8577, and Hayes, John D. Transcription factor Nrf2 mediates an adaptive response to sulforaphane that protects fibroblasts in vitro against the cytotoxic effects of electrophiles, peroxides and redox-cycling agents. United States: N. p., 2009. Web. doi:10.1016/j.taap.2009.03.005.
Higgins, Larry G, Kelleher, Michael O, Eggleston, Ian M, Itoh, Ken, Yamamoto, Masayuki, Department of Medical Biochemistry, Tohoku University Graduate School of Medicine, Sendai 980-8577, ERATO Environmental Response Project, Japan Science and Technology Corporation, Tsukuba 305-8577, & Hayes, John D. Transcription factor Nrf2 mediates an adaptive response to sulforaphane that protects fibroblasts in vitro against the cytotoxic effects of electrophiles, peroxides and redox-cycling agents. United States. https://doi.org/10.1016/j.taap.2009.03.005
Higgins, Larry G, Kelleher, Michael O, Eggleston, Ian M, Itoh, Ken, Yamamoto, Masayuki, Department of Medical Biochemistry, Tohoku University Graduate School of Medicine, Sendai 980-8577, ERATO Environmental Response Project, Japan Science and Technology Corporation, Tsukuba 305-8577, and Hayes, John D. 2009. "Transcription factor Nrf2 mediates an adaptive response to sulforaphane that protects fibroblasts in vitro against the cytotoxic effects of electrophiles, peroxides and redox-cycling agents". United States. https://doi.org/10.1016/j.taap.2009.03.005.
@article{osti_21272577,
title = {Transcription factor Nrf2 mediates an adaptive response to sulforaphane that protects fibroblasts in vitro against the cytotoxic effects of electrophiles, peroxides and redox-cycling agents},
author = {Higgins, Larry G and Kelleher, Michael O and Eggleston, Ian M and Itoh, Ken and Yamamoto, Masayuki and Department of Medical Biochemistry, Tohoku University Graduate School of Medicine, Sendai 980-8577 and ERATO Environmental Response Project, Japan Science and Technology Corporation, Tsukuba 305-8577 and Hayes, John D.},
abstractNote = {Sulforaphane can stimulate cellular adaptation to redox stressors through transcription factor Nrf2. Using mouse embryonic fibroblasts (MEFs) as a model, we show herein that the normal homeostatic level of glutathione in Nrf2{sup -/-} MEFs was only 20% of that in their wild-type counterparts. Furthermore, the rate of glutathione synthesis following its acute depletion upon treatment with 3 {mu}mol/l sulforaphane was very substantially lower in Nrf2{sup -/-} MEFs than in wild-type cells, and the rebound leading to a {approx} 1.9-fold increase in glutathione that occurred 12-24 h after Nrf2{sup +/+} MEFs were treated with sulforaphane was not observed in Nrf2{sup -/-} fibroblasts. Wild-type MEFs that had been pre-treated for 24 h with 3 {mu}mol/l sulforaphane exhibited between 1.4- and 3.2-fold resistance against thiol-reactive electrophiles, including isothiocyanates, {alpha},{beta}-unsaturated carbonyl compounds (e.g. acrolein), aryl halides and alkene epoxides. Pre-treatment of Nrf2{sup +/+} MEFs with sulforaphane also protected against hydroperoxides (e.g. cumene hydroperoxide, CuOOH), free radical-generating compounds (e.g. menadione), and genotoxic electrophiles (e.g. chlorambucil). By contrast, Nrf2{sup -/-} MEFs were typically {approx} 50% less tolerant of these agents than wild-type fibroblasts, and sulforaphane pre-treatment did not protect the mutant cells against xenobiotics. To test whether Nrf2-mediated up-regulation of glutathione represents the major cytoprotective mechanism stimulated by sulforaphane, 5 {mu}mol/l buthionine sulfoximine (BSO) was used to inhibit glutathione synthesis. In Nrf2{sup +/+} MEFs pre-treated with sulforaphane, BSO diminished intrinsic resistance and abolished inducible resistance to acrolein, CuOOH and chlorambucil, but not menadione. Thus Nrf2-dependent up-regulation of GSH is the principal mechanism by which sulforaphane pre-treatment induced resistance to acrolein, CuOOH and chlorambucil, but not menadione.},
doi = {10.1016/j.taap.2009.03.005},
url = {https://www.osti.gov/biblio/21272577}, journal = {Toxicology and Applied Pharmacology},
issn = {0041-008X},
number = 3,
volume = 237,
place = {United States},
year = {Mon Jun 15 00:00:00 EDT 2009},
month = {Mon Jun 15 00:00:00 EDT 2009}
}