title: GSK-3β downregulates Nrf2 in cultured cortical neurons and in a rat model of cerebral ischemia-reperfusion.
abstract: The NF-E2-related factor 2 (Nrf2)/antioxidant response element (ARE) pathway plays a critical role in protecting against oxidative stress in brain ischemia and reperfusion injury. Glycogen synthase kinase 3β (GSK-3β) may play a critical role in regulating Nrf2 in a Kelch-like ECH-associated protein 1 (Keap1)-independent manner. However, the relationship between GSK-3β and Nrf2 in brain ischemia and reperfusion injury is not clear. In this study, we explored the mechanisms through which GSK-3β regulates Nrf2 and Nrf-2/ARE pathways in vitro and in vivo. We used oxygen and glucose deprivation/reoxygenation (OGD/R) in primary cultured cortical neurons and a middle cerebral artery occlusion-reperfusion (MCAO/R) rat model to mimic ischemic insult. In this study, GSK-3β siRNA and inhibitors (SB216763 and LiCl) were used to inhibit GSK-3β in vitro and in vivo. After inhibiting GSK-3β, expression of total and nuclear Nrf2, Nrf2-ARE binding activity, and expression of Nrf2/ARE pathway-driven genes HO-1 and NQO-1 increased. Overexpression of GSK-3β yielded opposite results. These results suggest that GSK-3β downregulates Nrf2 and the Nrf2/ARE pathway in brain ischemia and reperfusion injury. GSK-3β may be an endogenous antioxidant relevant protein, and may represent a new therapeutic target in treatment of ischemia and reperfusion injury.
authors.affiliation: Department of Pathology, Chongqing Medical University, 400016 Chongqing, People's Republic of China.|Institute of Neuroscience, Chongqing Medical University, 400016 Chongqing, People's Republic of China.|Key Laboratory of Neurobiology, Chongqing Medical University, 400016 Chongqing, People's Republic of China.
authors.forename: Xi
authors.initials: X
authors.lastname: Chen
authors.affiliation: Department of Pathology, Chongqing Medical University, 400016 Chongqing, People's Republic of China.|Institute of Neuroscience, Chongqing Medical University, 400016 Chongqing, People's Republic of China.|Key Laboratory of Neurobiology, Chongqing Medical University, 400016 Chongqing, People's Republic of China.
authors.forename: Yuanling
authors.initials: Y
authors.lastname: Liu
authors.affiliation: Department of Pathology, Chongqing Medical University, 400016 Chongqing, People's Republic of China.|Institute of Neuroscience, Chongqing Medical University, 400016 Chongqing, People's Republic of China.|Key Laboratory of Neurobiology, Chongqing Medical University, 400016 Chongqing, People's Republic of China.
authors.forename: Jin
authors.initials: J
authors.lastname: Zhu
authors.affiliation: Department of Respiratory Medicine, Jiangjin Center Hospital, 402260, Chongqing, People's Republic of China.
authors.forename: Shipeng
authors.initials: S
authors.lastname: Lei
authors.affiliation: Department of Forensic Medicine, Chongqing Medical University, 400016 Chongqing, People's Republic of China.
authors.forename: Yuan
authors.initials: Y
authors.lastname: Dong
authors.affiliation: Department of Pathology, Chongqing Medical University, 400016 Chongqing, People's Republic of China.|Institute of Neuroscience, Chongqing Medical University, 400016 Chongqing, People's Republic of China.|Key Laboratory of Neurobiology, Chongqing Medical University, 400016 Chongqing, People's Republic of China.
authors.forename: Lingyu
authors.initials: L
authors.lastname: Li
authors.affiliation: Department of Pathology, Chongqing Medical University, 400016 Chongqing, People's Republic of China.|Institute of Neuroscience, Chongqing Medical University, 400016 Chongqing, People's Republic of China.|Key Laboratory of Neurobiology, Chongqing Medical University, 400016 Chongqing, People's Republic of China.
authors.forename: Beibei
authors.initials: B
authors.lastname: Jiang
authors.affiliation: Department of Pathology, Chongqing Medical University, 400016 Chongqing, People's Republic of China.|Institute of Neuroscience, Chongqing Medical University, 400016 Chongqing, People's Republic of China.|Key Laboratory of Neurobiology, Chongqing Medical University, 400016 Chongqing, People's Republic of China.
authors.forename: Li
authors.initials: L
authors.lastname: Tan
authors.affiliation: Department of Pathology, Chongqing Medical University, 400016 Chongqing, People's Republic of China.|Institute of Neuroscience, Chongqing Medical University, 400016 Chongqing, People's Republic of China.|Key Laboratory of Neurobiology, Chongqing Medical University, 400016 Chongqing, People's Republic of China.
authors.forename: Jingxian
authors.initials: J
authors.lastname: Wu
authors.affiliation: Department of Pathology, Chongqing Medical University, 400016 Chongqing, People's Republic of China.|Institute of Neuroscience, Chongqing Medical University, 400016 Chongqing, People's Republic of China.|Key Laboratory of Neurobiology, Chongqing Medical University, 400016 Chongqing, People's Republic of China.
authors.forename: Shanshan
authors.initials: S
authors.lastname: Yu
authors.affiliation: Department of Pathology, Chongqing Medical University, 400016 Chongqing, People's Republic of China.|Institute of Neuroscience, Chongqing Medical University, 400016 Chongqing, People's Republic of China.|Key Laboratory of Neurobiology, Chongqing Medical University, 400016 Chongqing, People's Republic of China.
authors.forename: Yong
authors.initials: Y
authors.lastname: Zhao
mesh_terms: D000818:Animals; D002545:Brain Ischemia / Q000235:genetics / Q000378:metabolism*; D002478:Cells, Cultured; D004195:Disease Models, Animal; D015536:Down-Regulation; D005786:Gene Expression Regulation / Q000187:drug effects; D020868:Gene Silencing; D000071679:Glycogen Synthase Kinase 3 beta / Q000235:genetics / Q000378:metabolism*; D007211:Indoles / Q000494:pharmacology; D018021:Lithium Chloride / Q000494:pharmacology; D008297:Male; D008301:Maleimides / Q000494:pharmacology; D051267:NF-E2-Related Factor 2 / Q000378:metabolism*; D009474:Neurons / Q000166:cytology* / Q000378:metabolism; D051381:Rats; D015398:Signal Transduction / Q000187:drug effects
chemical_list: D007211:Indoles; D008301:Maleimides; D051267:NF-E2-Related Factor 2; C495637:Nfe2l2 protein, rat; C417521:SB 216763; D000071679:Glycogen Synthase Kinase 3 beta; C000605508:Gsk3b protein, rat; D018021:Lithium Chloride
references.citation: Free Radic Biol Med. 2004 May 15;36(10):1199-207
references.pmid: 15110384
references.citation: Am J Physiol Heart Circ Physiol. 2008 Mar;294(3):H1497-500
references.pmid: 18223186
references.citation: J Pharmacol Exp Ther. 2006 Jul;318(1):124-31
references.pmid: 16611852
references.citation: Brain Res. 2010 Jan 13;1308:167-75
references.pmid: 19857470
references.citation: Glia. 2009 Apr 15;57(6):645-56
references.pmid: 18942756
references.citation: Neurosci Bull. 2012 Oct;28(5):509-16
references.pmid: 23054633
references.citation: J Neurochem. 2008 Apr;105(1):192-202
references.pmid: 18005231
references.citation: J Neurochem. 2006 Aug;98(4):1316-23
references.pmid: 16771826
references.citation: FASEB J. 2011 Mar;25(3):1076-87
references.pmid: 21097520
references.citation: Neuropharmacology. 2008 Sep;55(3):310-8
references.pmid: 18308346
references.citation: PLoS One. 2012;7(6):e40165
references.pmid: 22768247
references.citation: Genes Dev. 1999 Jan 1;13(1):76-86
references.pmid: 9887101
references.citation: Biomaterials. 2014 Mar;35(9):2720-9
references.pmid: 24406215
references.citation: Circ Res. 2009 Jun 5;104(11):1240-52
references.pmid: 19498210
references.citation: Toxicol Appl Pharmacol. 2010 Jan 1;242(1):18-28
references.pmid: 19781563
references.citation: Mol Cell Biol. 2003 Nov;23(22):8137-51
references.pmid: 14585973
references.citation: Free Radic Biol Med. 2012 Jan 15;52(2):473-87
references.pmid: 22142471
references.citation: J Biol Chem. 2011 Aug 19;286(33):28821-32
references.pmid: 21690096
references.citation: J Biol Chem. 2006 Apr 28;281(17):12132-42
references.pmid: 16513647
references.citation: Free Radic Biol Med. 2014 Feb;67:387-95
references.pmid: 24355211
references.citation: Molecules. 2011;16(11):9234-44
references.pmid: 22051934
references.citation: J Biol Chem. 2006 May 26;281(21):14841-51
references.pmid: 16551619
references.citation: Am J Physiol Cell Physiol. 2003 Oct;285(4):C899-911
references.pmid: 12814913
references.citation: Mol Cell Biol. 2011 Mar;31(6):1121-33
references.pmid: 21245377
references.citation: Oncogene. 2013 Aug 8;32(32):3765-81
references.pmid: 22964642
references.citation: Can J Physiol Pharmacol. 2010 Sep;88(9):907-17
references.pmid: 20921977
references.citation: J Mol Cell Cardiol. 2006 May;40(5):708-16
references.pmid: 16516918
references.citation: J Biol Chem. 2007 Jun 1;282(22):16502-10
references.pmid: 17403689
references.citation: Antioxid Redox Signal. 2009 Mar;11(3):497-508
references.pmid: 18717629
references.citation: Oncogene. 2001 Jun 28;20(29):3906-17
references.pmid: 11439354
references.citation: Proc Natl Acad Sci U S A. 2002 Sep 3;99(18):11908-13
references.pmid: 12193649
references.citation: Int J Neuropsychopharmacol. 2014 Apr;17(4):635-44
references.pmid: 24345484
references.citation: Front Mol Neurosci. 2012 Jan 30;5:1
references.pmid: 22319467
references.citation: Int J Alzheimers Dis. 2011;2011:985085
references.pmid: 21629716
references.citation: Circ Res. 2004 Apr 16;94(7):960-6
references.pmid: 14976126
references.citation: Adv Enzyme Regul. 2006;46:113-40
references.pmid: 16887173