Coordinate regulation of glutathione biosynthesis and release by Nrf2-expressing glia potently protects neurons from oxidative stress.
Astrocytes have a higher antioxidant potential in comparison to neurons. Pathways associated with this selective advantage include the transcriptional regulation of antioxidant enzymes via the action of the Cap'n'Collar transcription factor Nrf2 at the antioxidant response element (ARE). Here we show that Nrf2 overexpression can reengineer neurons to express this glial pathway and enhance antioxidant gene expression. However, Nrf2-mediated protection from oxidative stress is conferred primarily by glia in mixed cultures. The antioxidant properties of Nrf2-overexpressing glia are more pronounced than those of neurons, and a relatively small number of these glia (< 1% of total cell number added) could protect fully cocultured naive neurons from oxidative glutamate toxicity associated with glutathione (GSH) depletion. Microarray and biochemical analyses indicate a coordinated upregulation of enzymes involved in GSH biosynthesis (xCT cystine antiporter, gamma-glutamylcysteine synthetase, and GSH synthase), use (glutathione S-transferase and glutathione reductase), and export (multidrug resistance protein 1) with Nrf2 overexpression, leading to an increase in both media and intracellular GSH. Selective inhibition of glial GSH synthesis and the supplementation of media GSH indicated that an Nrf2-dependent increase in glial GSH synthesis was both necessary and sufficient for the protection of neurons, respectively. Neuroprotection was not limited to overexpression of Nrf2, because activation of endogenous glial Nrf2 by the small molecule ARE inducer, tert-butylhydroquinone, also protected against oxidative glutamate toxicity.
Shih
Andy Y
AY
Kinsmen Laboratory of Neurological Research, University of British Columbia, Vancouver, British Columbia, V6T 1Z3, Canada.
Johnson
Delinda A
DA
Wong
Gloria
G
Kraft
Andrew D
AD
Jiang
Lei
L
Erb
Heidi
H
Johnson
Jeffrey A
JA
Murphy
Timothy H
TH
D000256:Adenoviridae / Q000235:genetics; D000818:Animals; D000975:Antioxidants / Q000378:metabolism / Q000494:pharmacology; D017209:Apoptosis / Q000187:drug effects; D001643:Bridged Bicyclo Compounds / Q000378:metabolism; D016923:Cell Death / Q000187:drug effects; D002478:Cells, Cultured; D018920:Coculture Techniques; D004268:DNA-Binding Proteins / Q000096:biosynthesis* / Q000235:genetics; D004791:Enzyme Inhibitors / Q000494:pharmacology; D018698:Glutamic Acid / Q000633:toxicity; D005978:Glutathione / Q000031:analogs & derivatives* / Q000096:biosynthesis* / Q000378:metabolism / Q000494:pharmacology; D006801:Humans; D006873:Hydroquinones / Q000494:pharmacology; D051267:NF-E2-Related Factor 2; D009457:Neuroglia / Q000166:cytology / Q000378:metabolism*; D009474:Neurons / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D020411:Oligonucleotide Array Sequence Analysis; D018384:Oxidative Stress / Q000187:drug effects* / Q000502:physiology; D051381:Rats; D017208:Rats, Wistar; D020218:Response Elements; D019311:Staurosporine / Q000494:pharmacology; D015534:Trans-Activators / Q000096:biosynthesis* / Q000235:genetics
D000975:Antioxidants; D001643:Bridged Bicyclo Compounds; D004268:DNA-Binding Proteins; D004791:Enzyme Inhibitors; D006873:Hydroquinones; D051267:NF-E2-Related Factor 2; C495635:NFE2L2 protein, human; C495637:Nfe2l2 protein, rat; D015534:Trans-Activators; C080120:glutathione-bimane; D018698:Glutamic Acid; C018855:2-tert-butylhydroquinone; D005978:Glutathione; D019311:Staurosporine