Early Anesthetic Preconditioning in Mixed Cortical Neuronal-Glial Cell Cultures Subjected to Oxygen-Glucose Deprivation: The Role of Adenosine Triphosphate Dependent Potassium Channels and Reactive Oxygen Species in Sevoflurane-Induced Neuroprotection

Considering the wide, positive reporting of the role of reactive oxygen species in ischemic brain injury, searching for antioxidant drugs within herbal remedies is logical. In this study, the protective effects ofScutellaria litwinowiiBornm. & Sint. on cell viability and reactive oxygen species production in cultured PC12 cells were investigated under serum/glucose-deprivation-induced cell death. After cells were seeded overnight, they were then deprived of serum/glucose for 24 h. Cells were treated with different concentrations ofS. litwinowiiextract (7.75–250 μg/mL). Cell viability was quantitated by MTT assay, and intracellular reactive oxygen species production was measured by flow cytometry. Serum/glucose-deprivation induced significant cell death after 24 h (P< 0.001). Treatment withS. litwinowii(7.75–250 μg/mL) reduced serum/glucose deprivation-induced cytotoxicity in PC12 cells after 24 h. A significant increase in intracellular reactive oxygen species production was seen following serum/glucose deprivation (P< 0.001).S. litwinowii(62 and 125 μg/mL,P< 0.01) treatment reversed the increased reactive oxygen species production following ischemic insult. This demonstrates thatS. litwinowiiextract protects PC12 cells against serum/glucose-deprivation-induced cell death by antioxidant mechanisms, which indicates the potential therapeutic application ofS. litwinowiiin managing cerebral ischemic and neurodegenerative disorders.

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Sevoflurane Protects Rat Mixed Cerebrocortical Neuronal–Glial Cell Cultures against Transient Oxygen–Glucose Deprivation

Anesthesiology ◽

10.1097/00000542-200611000-00021 ◽

2006 ◽

Vol 105 (5) ◽

pp. 990-998 ◽

Cited By ~ 23

Author(s):

Paula T. Canas ◽

Lionel J. Velly ◽

Christelle N. Labrande ◽

Benjamin A. Guillet ◽

Valérie Sautou-Miranda ◽

...

Keyword(s):

Reactive Oxygen Species ◽

In Vitro Model ◽

Glutamate Uptake ◽

Reactive Oxygen ◽

Glucose Deprivation ◽

Oxygen Species ◽

Lactate Dehydrogenase Release ◽

Vitro Model ◽

Glial Cell Cultures

Background 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. Methods 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. Results 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. Conclusion 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.

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