title: Protective effect of 20-HETE inhibition in a model of oxygen-glucose deprivation in hippocampal slice cultures.
abstract: Recent studies have indicated that inhibitors of the synthesis of 20-hydroxyeicosatetraenoic acid (20-HETE) may have direct neuroprotective actions since they reduce infarct volume after ischemia reperfusion in the brain without altering blood flow. To explore this possibility, the present study used organotypic hippocampal slice cultures subjected to oxygen-glucose deprivation (OGD) and reoxygenation to examine whether 20-HETE is released by organotypic hippocampal slices after OGD and whether it contributes to neuronal death through the generation of ROS and activation of caspase-3. The production of 20-HETE increased twofold after OGD and reoxygenation. Blockade of the synthesis of 20-HETE with N-hydroxy-N'-(4-butyl-2-methylphenol)formamidine (HET0016) or its actions with a 20-HETE antagonist, 20-hydroxyeicosa-6(Z),15(Z)-dienoic acid, reduced cell death, as measured by the release of lactate dehydrogenase and propidium iodide uptake. Administration of a 20-HETE mimetic, 20-hydroxyeicosa-5(Z),14(Z)-dienoic acid (5,14-20-HEDE), had the opposite effect and increased injury after OGD. The death of neurons after OGD was associated with an increase in the production of ROS and activation of caspase-3. These effects were attenuated by HET0016 and potentiated after the administration of 5,14-20-HEDE. These findings indicate that the production of 20-HETE by hippocampal slices is increased after OGD and that inhibitors of the synthesis or actions of 20-HETE protect neurons from ischemic cell death. The protective effect of 20-HETE inhibitors is associated with a decrease in superoxide production and activation of caspase-3.
authors.affiliation: Department of Physiology, Medical College of Wisconsin, Milwaukee, USA.
authors.forename: Marija
authors.initials: M
authors.lastname: Renic
authors.forename: Suresh N
authors.initials: SN
authors.lastname: Kumar
authors.forename: Debebe
authors.initials: D
authors.lastname: Gebremedhin
authors.forename: Matthew A
authors.initials: MA
authors.lastname: Florence
authors.forename: Nashaat Z
authors.initials: NZ
authors.lastname: Gerges
authors.forename: John R
authors.initials: JR
authors.lastname: Falck
authors.forename: David R
authors.initials: DR
authors.lastname: Harder
authors.forename: Richard J
authors.initials: RJ
authors.lastname: Roman
mesh_terms: D000578:Amidines / Q000494:pharmacology*; D000818:Animals; D000831:Animals, Newborn; D053148:Caspase 3 / Q000378:metabolism; D016923:Cell Death / Q000187:drug effects; D015687:Cell Hypoxia; D019610:Cytoprotection; D005947:Glucose / Q000172:deficiency*; D006624:Hippocampus / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D006893:Hydroxyeicosatetraenoic Acids / Q000037:antagonists & inhibitors* / Q000378:metabolism / Q000494:pharmacology*; D018696:Neuroprotective Agents / Q000494:pharmacology*; D051381:Rats; D017207:Rats, Sprague-Dawley; D013481:Superoxides / Q000378:metabolism; D013997:Time Factors; D046509:Tissue Culture Techniques
chemical_list: C494665:20-hydroxyeicosa-5(Z),14(Z)-dienoic acid; C494667:20-hydroxyeicosa-6(Z),15(Z)-dienoic acid; D000578:Amidines; C000708209:HET0016; D006893:Hydroxyeicosatetraenoic Acids; D018696:Neuroprotective Agents; D013481:Superoxides; C055987:20-hydroxy-5,8,11,14-eicosatetraenoic acid; C505410:Casp3 protein, rat; D053148:Caspase 3; D005947:Glucose