title: Influence of post-hypoxia reoxygenation conditions on energy metabolism and superoxide production in cultured neurons from the rat forebrain.
abstract: Brain reperfusion and/or reoxygenation may be of particular importance in the etiology of neuronal damage after hypoxic-ischemic insult in neonates, especially with reference to the generation of free radicals. To investigate this issue, the influence of either standard reoxygenation or transient hyperoxia was studied on the consequences of severe hypoxia in a model of cultured neurons isolated from the fetal rat brain. Culture dishes were exposed for 6 h to hypoxia (95% N2/5% CO2). They were then placed under normoxia (95% air/5% CO2) or hyperoxia (95% O2/5% CO2) for 3 h, and finally returned to normoxia. Control cultures were kept under normoxic conditions. Cell morphology, protein concentrations, lactate dehydrogenase leakage, energy metabolism, as reflected by specific transport and incorporation of 2-D-[3H]deoxyglucose, as well as superoxide radical formation were analyzed as a function of time. Po2 values in the cell incubating medium were decreased by 78% by hypoxia and increased by 221% by hyperoxia. No morphologic alteration could be noticed before 72 h posthypoxia, when cell degeneration became apparent, with a concomitant reduction in protein contents. Hypoxia-reoxygenation induced a transient cellular hypermetabolism, as shown by a 36% increase in 2-D-[3H]deoxyglucose uptake 24 h after hypoxia, and then a 23% decrease below control values at 72 h. It also led to a sharp increase in the formation of superoxide radicals (+108%). Transient hyperoxia during reoxygenation did not exacerbate these events, and thus would not enhance their deterimental effects on cell integrity.
authors.affiliation: INSERM U.272, 54013 Nancy, France.
authors.forename: J
authors.initials: J
authors.lastname: Oillet
authors.forename: V
authors.initials: V
authors.lastname: Koziel
authors.forename: P
authors.initials: P
authors.lastname: Vert
authors.forename: J L
authors.initials: JL
authors.lastname: Daval
mesh_terms: D000818:Animals; D015687:Cell Hypoxia; D002478:Cells, Cultured; D003847:Deoxyglucose / Q000493:pharmacokinetics; D004734:Energy Metabolism; D005110:Extracellular Space / Q000378:metabolism; D005260:Female; D006801:Humans; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D008297:Male; D009474:Neurons / Q000166:cytology / Q000378:metabolism*; D010100:Oxygen / Q000378:metabolism*; D011247:Pregnancy; D016548:Prosencephalon / Q000166:cytology / Q000196:embryology; D051381:Rats; D017207:Rats, Sprague-Dawley; D015424:Reperfusion; D013481:Superoxides / Q000378:metabolism*
chemical_list: D013481:Superoxides; D003847:Deoxyglucose; D007770:L-Lactate Dehydrogenase; D010100:Oxygen