Abstract
Recent studies indicate that NF-E2 related factor 2 (Nrf2) is a substrate for the ubiquitin-proteasome pathway. The present study is aimed to determine whether increased protein stability is a mechanism by which quinone compounds, like tert-butylhydroquinone (tBHQ), may enhance Nrf2-mediated transcriptional activation and subsequent antioxidant protection. H2O2-induced necrotic cell death, evidenced by transmission electronic microscope (TEM) imaging with no caspase 3 activation and PARP cleavage, was significantly attenuated by pretreatment with tBHQ or overexpression of Nrf2 through advenovirus-mediated infection in human neural stem cells (hNSCs). Microarray analysis showed that those identified antioxidant genes, responsible for antiapoptotic action in IMR-32 cells (J. Li et al., 2002, J. Biol. Chem. 277, 388-394), were also coordinately upregulated through Nrf2-dependent antioxidant responsive element (ARE) activation in hNSC. The stabilization of Nrf2 by tBHQ in IMR-32 cells was evidenced by a pulse-chase assay showing no significant increase in Nrf2 protein synthesis after tBHQ treatment, and by ubiquitin immunoprecipitation showing that tBHQ stabilized ubiquitinated Nrf2. An in vitro proteasomal activity assay showed that tBHQ did not act as a 20S/26S proteasome inhibitor. Nrf2 stabilization by tBHQ also was observed in hNSCs. Taken together, this study suggests that identified antioxidant genes, which were upregulated. through tBHQ induced Nrf2 stabilization, confer protection on target cells against H2O2-induced apoptotic cell death in neuroblastoma cells as well as the necrotic cell death in the hNSC. Nrf2 stabilization by pharmacological modulation or adenovirus-mediated Nrf2 overexpression, therefore, might be viable strategies to prevent a wide-spectrum of oxidative stress-related neuronal cell injuries.
Original language | English (US) |
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Pages (from-to) | 313-328 |
Number of pages | 16 |
Journal | Toxicological Sciences |
Volume | 83 |
Issue number | 2 |
DOIs | |
State | Published - Feb 2005 |
Funding
We thank Microarray Core Facility of the University of Wisconsin Environmental Health Science Center for conducting the Affymetrix gene array hybridizations. We thank University of Wisconsin Medical School Electron Microscope Facility for TEM sample preparation. This study was supported by grants from the National Institutes of Environmental Health Sciences (ES08089, J.J.; ES10042, J.J.; ES09090, Environmental Health Sciences Center), the Wellcome Trust (C.S.), and Michael J. Fox Foundation (C.S.).
Keywords
- Apoptotic cell death
- Human neural stem cells
- Nrf2
- Tert-butylhydroquinone
ASJC Scopus subject areas
- Toxicology