Objective To investigate the ability of once and twice isoflurane preconditioning against oxygen and glucose deprivation（OGD） injury in rats brain in vitro.Methods Rat hippocampal slices were exposed to 1vol%,2vol% and 3vol% isoflurane for 30 min under normoxic condition（95% O2/5% CO2） once and twice（n=12 for each group） respectively before OGD.At the end of each exposure,the slices were set with a 15-min washout period interspersed,then slices were exposed to 13-min OGD period（95% N2/5%CO2,glucose-free） followed by 30min reoxygenation.The amplitude CA1 population spikes（PS,neuronal function） was measured and used to quantify the degree of recovery of neuronal function at post OGD period.To assess the role of the mitogen-activated protein kinases（MAPKs） in preconditioning,U0126,an inhibitor of extracellular signal-regulated protein kinase（MEK-ERK1/2）,and SB203580,an inhibitor of p38 MAPK,were used during isoflurane exposure.Results Isoflurane-preconditioning with 1vol%,2vol% and 3vol% once increased the degree of recovery from 4.8%±1.4%（control） to 41.9%±9.2%,55.1%±11.0% and 63.2%±10.8%,respectively,and twice to 53.8%±12.0%,63.5%±11.1% and 76.3%±12.3%, respectively.The effect of twice exposure to 3vol% isoflurane was blocked by U0126（6.1%±1.5%）.Conclusion It is concluded that twice isoflurane preconditioning could achieve better neuroprotection than once via activation of extracellular signal-regulated protein kinase（MEK-ERK1/2）.
利用离体海马脑片缺氧无糖(oxygen-glucose deprivation,OGD)损伤模型,探讨七氟醚预处理对神经细胞的保护作用及陔作用与线粒体内膜ATP敏感钾通道(mitochondrial ATP-sensitive potassium channels,mitoK_(ATP)channels)的关系,随机将脑片用2%、4%、6%七氟醚,以及6%七氟醚复合mitoK_(ATP)通道阻滞剂5-羟基奎酸盐(5-hydroxydecanoic acid,5-HD)预处理30min,观察OGD损伤14min复氧1h期间顺向群峰电位(orthodromic population spike,OPS)的变化,并应用透射电镜观察细胞超微结构的改变。结果表明,与单纯OGD组相比,七氟醚预处理可使海马脑片OPS消失时间明显延长(P＜0．01),使OPS明显恢复,其中4%、6%七氟醚组的恢复率均为71．4%(P＜0．05 vs OGD),相应恢复程度为(61．0±42．3)%和(78．7±21．1)%(P＜0．01),而且6%七氟醚的保护作用可被5-HD取消。OGD组的海马CA1区锥体细胞明显水肿,核膜皱缩、破裂,染色质聚集,线粒体肿胀畸形,嵴断裂或消失,而4%和6%七氟醚组仅见海马CA1区锥体细胞轻度水肿,核膜皱缩不明显,染色质均匀,线粒体轻度肿胀。结果提示,七氟醚预处理对大鼠海马脑片OGD损伤有一定的保护作用,且七氟醚对神经细胞的保护作用与激活mitoK_(ATP)通道有关。
