Sevoflurane protects rat mixed cerebrocortical neuronal-glial cell cultures against transient oxygen-glucose deprivation : Involvement of glutamate uptake and reactive oxygen species

The purpose of this study was to clarify the role of glutamate and reactive oxygen species in sevoflurane-mediated neuroprotection on an in vitro model of ischemia-reoxygenation. Mature mixed cerebrocortical neuronal-glial cell cultures, treated or not with increasing concentrations of sevoflurane,...

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Bibliographic Details
Published in:Anesthesiology (Philadelphia) 2006-11, Vol.105 (5), p.990-998
Main Authors: CANAS, Paula T, VELLY, Lionel J, LABRANDE, Christelle N, GUILLET, Benjamin A, SAUTOU-MIRANDA, Valérie, MASMEJEAN, Frédérique M, NIEOULLON, André L, GOUIN, Francois M, BRUDER, Nicolas J, PISANO, Pascale S
Format: Article
Language:English
Subjects:
Anesthesia
Anesthesia. Intensive care medicine. Transfusions. Cell therapy and gene therapy
Animals
Biological and medical sciences
Biological Transport, Active - drug effects
Cell Hypoxia - drug effects
Cells, Cultured
Cellular Biology
Cerebral Cortex - cytology
Cerebral Cortex - drug effects
Cerebral Cortex - metabolism
Coculture Techniques
Glucose - metabolism
Glutamic Acid - metabolism
Life Sciences
Medical sciences
Methyl Ethers - pharmacology
Neuroglia - drug effects
Neuroglia - metabolism
Neurons - drug effects
Neurons - metabolism
Neuroprotective Agents - pharmacology
Rats
Reactive Oxygen Species - metabolism
Sevoflurane
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Summary:The purpose of this study was to clarify the role of glutamate and reactive oxygen species in sevoflurane-mediated neuroprotection on an in vitro model of ischemia-reoxygenation. Mature mixed cerebrocortical neuronal-glial cell cultures, treated or not with increasing concentrations of sevoflurane, were exposed to 90 min combined oxygen-glucose deprivation (OGD) in an anaerobic chamber followed by reoxygenation. Cell death was quantified by lactate dehydrogenase release into the media and cell viability by reduction of 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium by mitochondrial succinate dehydrogenase. Extracellular concentrations of glutamate and glutamate uptake were assessed at the end of the ischemic injury by high-performance liquid chromatography and incorporation of L-[H]glutamate into cells, respectively. Free radical generation in cells was assessed 6 h after OGD during the reoxygenation period using 2',7'-dichlorofluorescin diacetate, which reacts with intracellular radicals to be converted to its fluorescent product, 2',7'-dichlorofluorescin, in cell cytosol. Twenty-four hours after OGD, sevoflurane, in a concentration-dependent manner, significantly reduced lactate dehydrogenase release and increased cell viability. At the end of OGD, sevoflurane was able to reduce the OGD-induced decrease in glutamate uptake. This effect was impaired in the presence of threo-3-methyl glutamate, a specific inhibitor of the glial transporter GLT1. Sevoflurane counteracted the increase in extracellular level of glutamate during OGD and the generation of reactive oxygen species during reoxygenation. Sevoflurane had a neuroprotective effect in this in vitro model of ischemia-reoxygenation. This beneficial effect may be explained, at least in part, by sevoflurane-induced antiexcitotoxic properties during OGD, probably depending on GLT1, and by sevoflurane-induced decrease of reactive oxygen species generation during reoxygenation.
ISSN:0003-3022
1528-1175
DOI:10.1097/00000542-200611000-00021