title: Sulforaphane preconditioning of the Nrf2/HO-1 defense pathway protects the cerebral vasculature against blood-brain barrier disruption and neurological deficits in stroke.
abstract: Disruption of the blood-brain barrier (BBB) and cerebral edema are the major pathogenic mechanisms leading to neurological dysfunction and death after ischemic stroke. The brain protects itself against infarction via activation of endogenous antioxidant defense mechanisms, and we here report the first evidence that sulforaphane-mediated preactivation of nuclear factor erythroid 2-related factor 2 (Nrf2) and its downstream target heme oxygenase-1 (HO-1) in the cerebral vasculature protects the brain against stroke. To induce ischemic stroke, Sprague-Dawley rats were subjected to 70 min middle cerebral artery occlusion (MCAo) followed by 4, 24, or 72 h reperfusion. Nrf2 and HO-1 protein expression was upregulated in cerebral microvessels of peri-infarct regions after 4-72 h, with HO-1 preferentially associated with perivascular astrocytes rather than the cerebrovascular endothelium. In naïve rats, treatment with sulforaphane increased Nrf2 expression in cerebral microvessels after 24h. Upregulation of Nrf2 by sulforaphane treatment prior to transient MCAo (1h) was associated with increased HO-1 expression in perivascular astrocytes in peri-infarct regions and cerebral endothelium in the infarct core. BBB disruption, lesion progression, as analyzed by MRI, and neurological deficits were reduced by sulforaphane pretreatment. As sulforaphane pretreatment led to a moderate increase in peroxynitrite generation, we suggest that hormetic preconditioning underlies sulforaphane-mediated protection against stroke. In conclusion, we propose that pharmacological or dietary interventions aimed to precondition the brain via activation of the Nrf2 defense pathway in the cerebral microvasculature provide a novel therapeutic approach for preventing BBB breakdown and neurological dysfunction in stroke.
authors.affiliation: Cardiovascular Division, British Heart Foundation Centre of Research Excellence, School of Medicine, King's College London, London SE1 9NH, UK.
authors.forename: Alessio
authors.initials: A
authors.lastname: Alfieri
authors.affiliation: Cardiovascular Division, British Heart Foundation Centre of Research Excellence, School of Medicine, King's College London, London SE1 9NH, UK.
authors.forename: Salil
authors.initials: S
authors.lastname: Srivastava
authors.affiliation: Cardiovascular Division, British Heart Foundation Centre of Research Excellence, School of Medicine, King's College London, London SE1 9NH, UK.
authors.forename: Richard C M
authors.initials: RCM
authors.lastname: Siow
authors.affiliation: Centre for Neuroimaging Sciences, Institute of Psychiatry, King's College London, London SE5 8AF, UK.
authors.forename: Diana
authors.initials: D
authors.lastname: Cash
authors.affiliation: Centre for the Cellular Basis of Behaviour, Institute of Psychiatry, King's College London, London SE5 9NU, UK.
authors.forename: Michel
authors.initials: M
authors.lastname: Modo
authors.affiliation: Department of Cell and Developmental Biology, University College London, London WC1E 6BT, UK.
authors.forename: Michael R
authors.initials: MR
authors.lastname: Duchen
authors.affiliation: Cardiovascular Division, British Heart Foundation Centre of Research Excellence, School of Medicine, King's College London, London SE1 9NH, UK.
authors.forename: Paul A
authors.initials: PA
authors.lastname: Fraser
authors.affiliation: Centre for Neuroimaging Sciences, Institute of Psychiatry, King's College London, London SE5 8AF, UK.
authors.forename: Steven C R
authors.initials: SCR
authors.lastname: Williams
authors.affiliation: Cardiovascular Division, British Heart Foundation Centre of Research Excellence, School of Medicine, King's College London, London SE1 9NH, UK. Electronic address: giovanni.mann@kcl.ac.uk.
authors.forename: Giovanni E
authors.initials: GE
authors.lastname: Mann
mesh_terms: D000818:Animals; D001812:Blood-Brain Barrier / Q000187:drug effects / Q000378:metabolism* / Q000473:pathology; D002536:Cerebral Arteries / Q000187:drug effects / Q000201:enzymology; D015870:Gene Expression; D051547:Heme Oxygenase-1 / Q000235:genetics / Q000378:metabolism*; D020244:Infarction, Middle Cerebral Artery / Q000188:drug therapy / Q000378:metabolism*; D017879:Isothiocyanates / Q000494:pharmacology* / Q000627:therapeutic use; D008297:Male; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism*; D055806:Microvessels / Q000201:enzymology; D051267:NF-E2-Related Factor 2 / Q000235:genetics / Q000378:metabolism*; D018696:Neuroprotective Agents / Q000494:pharmacology* / Q000627:therapeutic use; D018384:Oxidative Stress; D011596:Psychomotor Disorders / Q000378:metabolism / Q000517:prevention & control*; D017207:Rats, Sprague-Dawley; D015427:Reperfusion Injury / Q000378:metabolism / Q000517:prevention & control; D015398:Signal Transduction; D013454:Sulfoxides; D015854:Up-Regulation
chemical_list: D017879:Isothiocyanates; D008565:Membrane Proteins; D051267:NF-E2-Related Factor 2; D018696:Neuroprotective Agents; C495636:Nfe2l2 protein, mouse; D013454:Sulfoxides; D051547:Heme Oxygenase-1; C496542:Hmox1 protein, mouse; C016766:sulforaphane
keywords: 3,3′-diaminobenzidine; 4,6-diamidino-2-phenylindole; ARE; BBB; Blood–brain barrier; DAB; DAPI; GFAP; Gliovascular complex; HO; Heme oxygenase; Iba1; Keap1; Kelch-like ECH associated protein 1; MCAo; Nrf2; Preconditioning; Prx1; RECA-1; RNS; ROS; Stroke; Sulforaphane; antioxidant response element; blood–brain barrier; glial fibrillary acidic protein; heme oxygenase; ionized calcium binding adaptor molecule 1; middle cerebral artery occlusion; nuclear factor erythroid 2-related factor 2; peroxiredoxin 1; rat endothelial cell antigen-1; reactive nitrogen species; reactive oxygen species