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
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
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DOI：
10.1213/ane.0b013e318193fee7
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发表时间：
2009-03-01
影响因子：
5.7
通讯作者：
Pisano, Pascale S.
中科院分区：
文献类型：
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作者：
Velly, Lionel J.;Canas, Paula T.;Pisano, Pascale S.
背景：本研究的目的是对皮层神经元-子代混合细胞进行短暂氧-葡萄糖剥夺（OGD）培养：i)比较七氟醚在预处理前和直接神经保护期间的神经保护作用；ii)探讨三磷酸腺苷敏感钾（KATP）通道和细胞内活性氧（ROS）水平可能参与七氟醚早期预处理作用的机制。方法：将成熟的混合皮层神经元-孝细胞培养物在厌氧室中暴露90分钟OGD，然后再氧化。七氟醚（0.03-3.4 mM）随机给药90分钟，在OGD前60分钟（早期预处理）或在OGD 90分钟（直接神经保护）期间停用。通过乳酸脱氢酶释放到沐浴培养基中，定量OGD后24 h细胞死亡情况。在七氟醚预处理结束时，用2',7'-双乙酸二氯荧光素评估细胞内RCS的产生。结果：浓度高于0.07 mM时，七氟醚预处理可引起阈值依赖性的神经保护作用，在OGD过程中，七氟醚的加入可引起剂量依赖性的神经保护作用。KATP通道阻滞剂（格列本脲0.3 μ M和5羟基癸酸50 μ M）或ros清除剂（n -2-巯基丙酰甘氨酸700 μ M和n -乙酰半胱氨酸50 μ M）虽然不影响细胞活力，但它们抵消了早期七氟醚预处理产生的神经保护作用。在预处理期间暴露于七氟醚诱导ROS水平显著增加，这被ROS清除剂和KATP通道阻滞剂阻止。结论：早期七氟醚预处理诱导混合皮质神经元-细胞培养物对OGD具有阈值依赖性保护作用，其机制似乎涉及打开KATP通道，从而导致ROS的产生。
BACKGROUND: The purpose of the present study, on mixed cortical neuronal-filial 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-filial 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 RCS 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 mu M and 5 hydroxydecanoic acid 50 AM), or ROS-scavengers (N-2-mercaptopropionyl glycine 700 AM and N-acetylcysteine 50 mu M), 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-filial cell cultures against OGD by mechanisms that seem to involve opening KATP channels, thereby leading to generation of ROS.