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NEUROSURGICAL ANESTHESIOLOGY AND NEUROSCIENCE

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

Lionel J. Velly, MD^*, Paula T. Canas, MD^*, Benjamin A. Guillet, PhD^;, Christelle N. Labrande, PhD, Frédérique M. Masmejean, BS, André L. Nieoullon, PhD, François M. Gouin, MD^*, Nicolas J. Bruder, MD^*, and Pascale S. Pisano, PhD

From the *Department of Anesthesia, Centre Hospitalier Universitaire Timone Adultes, Marseille, France; Laboratory of Pharmacodynamic, UMR INSERM 608, Marseille, France; and Laboratory IC2N, UMR6186 CNRS, Université de la Méditerranée, Marseille, France.

Address correspondence and reprint requests to Dr. Lionel Velly, Service d’AnesthésieRéanimation, Centre Hospitalier Universitaire Timone, Marseille, France. Address e-mail to lionel.velly{at}ap-hm.fr.

Abstract

BACKGROUND: The purpose of the present study, on mixed cortical neuronal-glial cell cultures subjected to transient oxygen-glucose deprivation (OGD) was: i) to compare the neuroprotection afforded by sevoflurane added either before (preconditioning) or during (direct neuroprotection) the OGD and ii) to explore the possible involvement of adenosine triphosphate-sensitive potassium (KATP) channels and intracellular reactive oxygen species (ROS) levels in the mechanism of the early preconditioning effect of sevoflurane.

METHODS: Mature mixed cortical neuronal-glial cell cultures were exposed to 90-min OGD in an anaerobic chamber followed by reoxygenation. Sevoflurane (0.03–3.4 mM) was randomly administered for 90 min and discontinued 60 min before OGD (early preconditioning) or during the 90-min OGD (direct neuroprotection). Cell death was quantified 24 h after the OGD by lactate dehydrogenase release into the bathing medium. Intracellular ROS generation was assessed at the end of sevoflurane preconditioning using 2',7'-dichlorofluorescin diacetate.

RESULTS: Sevoflurane preconditioning elicited a potent threshold-dependent neuroprotective effect at concentrations higher than 0.07 mM and sevoflurane added during OGD elicited a dose dependent neuroprotective effect. Blockers of KATP channels (glibenclamide 0.3 µM and 5 hydroxydecanoic acid 50 µM), or ROS-scavengers (N-2-mercaptopropionyl glycine 100 µM and N-acetylcysteine 50 µM), although they did not affect cell viability, counteracted the neuroprotection produced by early sevoflurane preconditioning. Sevoflurane exposure during preconditioning induced a significant increase in ROS levels which was prevented by both ROS scavengers and blockers of KATP channels.

CONCLUSION: Early sevoflurane preconditioning induced a threshold-dependent protection of mixed cortical neuronal-glial cell cultures against OGD by mechanisms that seem to involve opening KATP channels, thereby leading to generation of ROS.