title: Neuroprotective effect of an angiotensin receptor type 2 agonist following cerebral ischemia in vitro and in vivo.
abstract: BACKGROUND
  Intracerebral administration of the angiotensin II type 2 receptor (AT2R) agonist, CGP42112, is neuroprotective in a rat model of ischemic stroke. To explore further its possible cellular target(s) and therapeutic utility, we firstly examined whether CGP42112 may exert direct protective effects on primary neurons following glucose deprivation in vitro. Secondly, we tested whether CGP42112 is effective when administered systemically in a mouse model of cerebral ischemia.
  
  
  METHODS
  Primary cortical neurons were cultured from E17 C57Bl6 mouse embryos for 9 d, exposed to glucose deprivation for 24 h alone or with drug treatments, and percent cell survival assessed using trypan blue exclusion. Ischemic stroke was induced in adult male C57Bl6 mice by middle cerebral artery occlusion for 30 min, followed by reperfusion for 23.5 h. Neurological assessment was performed and then mice were euthanized and infarct and edema volume were analysed.
  
  
  RESULTS
  During glucose deprivation, CGP42112 (1x10-8 M and 1x10-7 M) reduced cell death by ~30%, an effect that was prevented by the AT2R antagonist, PD123319 (1x10-6 M). Neuroprotection by CGP42112 was lost at a higher concentration (1x10-6 M) but was unmasked by co-application with the AT1R antagonist, candesartan (1x10-7 M). By contrast, Compound 21 (1x10-8 M to 1x10-6 M), a second AT2R agonist, had no effect on neuronal survival. Mice treated with CGP42112 (1 mg/kg i.p.) after cerebral ischemia had improved functional outcomes over vehicle-treated mice as well as reduced total and cortical infarct volumes.
  
  
  CONCLUSIONS
  These results indicate that CGP42112 can directly protect neurons from ischemia-like injury in vitro via activation of AT2Rs, an effect opposed by AT1R activation at high concentrations. Furthermore, systemic administration of CGP42112 can reduce functional deficits and infarct volume following cerebral ischemia in vivo.
authors.affiliation: Department of Pharmacology, Monash University, Clayton, VIC, 3800, Australia.
authors.forename: Seyoung
authors.initials: S
authors.lastname: Lee
authors.affiliation: Department of Pharmacology, Monash University, Clayton, VIC, 3800, Australia.
authors.forename: Vanessa H
authors.initials: VH
authors.lastname: Brait
authors.affiliation: School of Biomedical Sciences, The University of Queensland, Brisbane, QLD, 4072, Australia.
authors.forename: Thiruma V
authors.initials: TV
authors.lastname: Arumugam
authors.affiliation: Department of Pharmacology, Monash University, Clayton, VIC, 3800, Australia.
authors.forename: Megan A
authors.initials: MA
authors.lastname: Evans
authors.affiliation: Department of Pharmacology, Monash University, Clayton, VIC, 3800, Australia.
authors.forename: Hyun Ah
authors.initials: HA
authors.lastname: Kim
authors.affiliation: Department of Pharmacology, Monash University, Clayton, VIC, 3800, Australia.
authors.forename: Robert E
authors.initials: RE
authors.lastname: Widdop
authors.affiliation: Department of Pharmacology, Monash University, Clayton, VIC, 3800, Australia.
authors.forename: Grant R
authors.initials: GR
authors.lastname: Drummond
authors.affiliation: Department of Pharmacology, Monash University, Clayton, VIC, 3800, Australia.
authors.forename: Christopher G
authors.initials: CG
authors.lastname: Sobey
authors.affiliation: Department of Pharmacology, Monash University, Clayton, VIC, 3800, Australia.
authors.forename: Emma S
authors.initials: ES
authors.lastname: Jones