title: Post-ischaemic mild hypothermia inhibits apoptosis in the penumbral region by reducing neuronal nitric oxide synthase activity and thereby preventing endothelin-1-induced hydroxyl radical formation.
abstract: Previously, we showed that treatment with resuscitative, post-ischaemic mild hypothermia (34 degrees C for 2 h) reduced apoptosis in the penumbra (cortex), but not in the core (striatum) of an endothelin-1 (Et-1)-induced focal cerebral infarct in the anaesthetized rat. Therefore, the purpose of this study was to investigate by which pathways resuscitative mild hypothermia exerts its neuroprotective effect in this model. The amino acids glutamate, serine, glutamine, alanine, taurine, arginine and the NO-related compound citrulline were sampled from the striatum and cortex of the ischaemic hemisphere using in vivo microdialysis. The in vivo salicylate trapping method was applied for monitoring hydroxyl radical formation via 2,3 dihydroxybenzoic acid (2,3 DHBA) detection. Caspase-3, neuronal nitric oxide synthase (nNOS) immunoreactivity and the volume of ischaemic damage were determined 24 h after the insult. In both the striatum and the cortex, Et-1-induced increases in glutamate, taurine and alanine were refractory to mild hypothermia. However, mild hypothermia significantly attenuated the ischaemia-induced 2,3 DHBA levels and the nNOS immunoreactivity in the cortex, but not in the striatum. These observations were associated with a decreased caspase-3 immunoreactivity. These results suggest that mild hypothermia exerts its neuroprotective effect in the penumbra partially by reducing nNOS activity and thereby preventing oxidative stress. Furthermore, we confirm our previous findings that the neuroprotective effect of resuscitative hypothermia is not mediated by changes in ischaemia-induced amino acid release as they could not be associated with the ischaemia-induced damage in the Et-1 rat model.
authors.affiliation: Department of Pharmaceutical Chemistry and Drug Analysis, Research Group Experimental Pharmacology, Vrije Universiteit Brussel, Laarbeeklaan 103, 1090 Brussels, Belgium.
authors.forename: An
authors.initials: A
authors.lastname: Van Hemelrijck
authors.forename: Said
authors.initials: S
authors.lastname: Hachimi-Idrissi
authors.forename: Sophie
authors.initials: S
authors.lastname: Sarre
authors.forename: Guy
authors.initials: G
authors.lastname: Ebinger
authors.forename: Yvette
authors.initials: Y
authors.lastname: Michotte
mesh_terms: D000596:Amino Acids / Q000378:metabolism; D000818:Animals; D017209:Apoptosis / Q000502:physiology*; D001831:Body Temperature / Q000502:physiology; D002545:Brain Ischemia / Q000150:complications / Q000201:enzymology / Q000473:pathology*; D053148:Caspase 3; D020169:Caspases / Q000378:metabolism; D002540:Cerebral Cortex / Q000201:enzymology / Q000473:pathology*; D002544:Cerebral Infarction / Q000473:pathology; D019332:Endothelin-1 / Q000494:pharmacology*; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005110:Extracellular Space / Q000378:metabolism; D017665:Hydroxyl Radical / Q000378:metabolism*; D007035:Hypothermia / Q000201:enzymology / Q000209:etiology / Q000473:pathology*; D007150:Immunohistochemistry; D008297:Male; D017551:Microdialysis; D019331:NG-Nitroarginine Methyl Ester / Q000494:pharmacology; D017072:Neostriatum / Q000201:enzymology / Q000378:metabolism / Q000473:pathology; D016877:Oxidants; D051381:Rats; D017208:Rats, Wistar
chemical_list: D000596:Amino Acids; D019332:Endothelin-1; D004791:Enzyme Inhibitors; D016877:Oxidants; D017665:Hydroxyl Radical; C505410:Casp3 protein, rat; D053148:Caspase 3; D020169:Caspases; D019331:NG-Nitroarginine Methyl Ester