title: Reduction of cerebral infarct volume by apocynin requires pretreatment and is absent in Nox2-deficient mice.
abstract: OBJECTIVE
  Reactive oxygen species (ROS) derived from Nox2-containing reduced form of nicotinamide adenine dinucleotide phosphate (NADPH) oxidase activity is reportedly detrimental in cerebrovascular disease. However, ROS generation by other Nox isoforms may have a physiological role. No Nox2-selective inhibitors have yet been identified, and thus it is unclear whether isoform non-selective Nox inhibitors would necessarily improve outcome after stroke. We assessed the effect of apocynin on cerebrovascular ROS production and also on outcome following cerebral ischaemia when administered either before ischaemia or after cerebral reperfusion. The involvement of Nox2-containing NADPH oxidase in the effects of apocynin was assessed using Nox2(-/-) mice.
  
  
  METHODS
  Transient cerebral ischaemia was induced by 0.5 h middle cerebral artery occlusion followed by 23.5 h reperfusion. Mice received apocynin (2.5 mg.kg(-1), i.p.) either 0.5 h before ischaemia or 1 h after reperfusion. In situ superoxide production after cerebral ischaemia-reperfusion was measured in brain sections of wild-type mice at 24 h using dihydroethidium fluorescence.
  
  
  RESULTS
  Treatment with apocynin 0.5 h before ischaemia reduced total infarct volume, neurological impairment and mortality in wild-type but not Nox2(-/-) mice. Conversely, treatment with apocynin 1 h after initiation of reperfusion had no protective effect. Cerebral ischaemia and reperfusion increased superoxide production in the brain at 24 h, and pretreatment but not posttreatment with apocynin reduced superoxide levels.
  
  
  CONCLUSIONS
  Apocynin improves outcome following stroke when administered before ischaemia in wild-type but not Nox2(-/-) mice.
authors.affiliation: Department of Pharmacology, Monash University, Clayton, VIC, Australia.
authors.forename: K A
authors.initials: KA
authors.lastname: Jackman
authors.forename: A A
authors.initials: AA
authors.lastname: Miller
authors.forename: T M
authors.initials: TM
authors.lastname: De Silva
authors.forename: P J
authors.initials: PJ
authors.lastname: Crack
authors.forename: G R
authors.initials: GR
authors.lastname: Drummond
authors.forename: C G
authors.initials: CG
authors.lastname: Sobey
mesh_terms: D000098:Acetophenones / Q000008:administration & dosage / Q000627:therapeutic use*; D000818:Animals; D001921:Brain / Q000187:drug effects / Q000378:metabolism; D004334:Drug Administration Schedule; D020244:Infarction, Middle Cerebral Artery / Q000150:complications / Q000378:metabolism / Q000517:prevention & control*; D002546:Ischemic Attack, Transient / Q000150:complications / Q000378:metabolism / Q000517:prevention & control*; D008297:Male; D008562:Membrane Glycoproteins / Q000172:deficiency* / Q000235:genetics; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D000074662:NADPH Oxidase 2; D019255:NADPH Oxidases / Q000172:deficiency* / Q000235:genetics; D017382:Reactive Oxygen Species / Q000378:metabolism
chemical_list: D000098:Acetophenones; D008562:Membrane Glycoproteins; D017382:Reactive Oxygen Species; C056165:acetovanillone; C488842:Cybb protein, mouse; D000074662:NADPH Oxidase 2; D019255:NADPH Oxidases