七氟醚预处理对谷氨酸诱发新生大鼠海马神经元钙离子释放的影响  

Effects of sevoflurane preconditioning on glutamate-induced intracellular calcium release in cultured newborn rat hippocampal neurons

作　　者：王秋筠[1] 李克忠[2] 姚尚龙[1] 李志华[3] 

机构地区：[1]华中科技大学同济医学院附属协和医院麻醉科,武汉市430022 [2]山东大学第二医院麻醉科 [3]河北医科大学第三医院麻醉科

出　　处：《中华麻醉学杂志》2008年第1期72-74,共3页Chinese Journal of Anesthesiology

摘　　要：目的通过评价七氟醚预处理对谷氨酸诱发新生大鼠海马神经元钙离子释放的影响，探讨七氟醚脑保护作用的机制。方法取出生24hWistar大鼠的原代培养海马神经元，培养12d后，随机分为4组（n=80）：对照组（C组）、谷氨酸组（G组）、谷氨酸＋1MAC七氟醚组（GM1组）和谷氨酸＋2MAC七氟醚组（GM2组），每组又分为细胞外液有钙和无钙2个亚组（n=40）。G组荧光染色后用1mmol／L谷氨酸刺激5min；GM1组和GM2组分别以1MAC、2MAC七氟醚处理1h后，再行荧光染色和谷氨酸刺激。用显微荧光技术测定海马神经元内游离钙离子浓度（[Ca^2＋]i）。结果G组、GM1组和GM2组[Ca^2＋]i最大值高于基础值，且高于C组（P〈0．01）；与G组比较，GM2组的2个亚组[Ca^2＋]i最大值均降低，在细胞外液无钙条件下GM1组[Ca^2＋]i最大值降低（P〈0．01）；与GM1组比较，GM2组的2个亚组[Ca^2＋]i最大值均降低（P〈0．01）。结论七氟醚预处理可能通过抑制谷氨酸诱发新生大鼠海马神经元[Ca^2＋]i升高，抑制钙超载。Objective To evaluate the effects sevoflurane preconditioning on glutamate-induced intracellular calcium release in cultured newborn rat hippocampal neurons. Methods Hippocampal neurons were enzymatically isolated from newborn rats at 24 h after birth and cultured for 12 days. The primarily cultured neurons were randomly divided into 4 groups： group Ⅰ control （C）; group Ⅱglutamate （G）; group Ⅲ 1 MAC sevoflurane ＋ glutamate （GM1） and group Ⅳ 2 MAC sevoflurane ＋ glutamate （GM2）. Each group was further divided into 2 subgroups： in one subgroup the extracellular HEPES-buffered salt solution contained CaCI： （subgroupCa^2＋ ） while in another subgroup CaCl2 was replaced by MgCl2 （ subgroup Mg^2＋ ）. Group C received no treatment of any kind; glutamate group was exposed to glutamate 1 mmol/L for 5 min after fluorescent staining; group GM1 and GM2 were exposed to 1 and 2 MAC sevoflurane respectively for 1 h before fluorescent staining and glutamate treatment. Microfluorescent technique was used to detect the intracellular free （iomized） Ca^2＋ concentration （ [Ca^2＋] i） in hippocampal neurons. Results The maximum [Ca^2＋]i was significantly increased in group G, GM1 and GM： as compared to the baseline values or group C （ P 〈 0.01 ）. The maximum [ Ca^2＋] i was significantly lower in group GM1 and GM： than in group G with CaCI： in extracellular solution. The maximum [ Ca^2＋ ]i was significantly lower in group GM2 than in group GM1 with and without CaCl2 in the extracellular solution. Conclusion Sevoflurane preconditioning can inhibit glutamate-induced calcium overload by decreasing [Ca^2＋ ]i in cultured newborn rat hippocampal neurons.

关 键 词：钙 海马 神经元 谷氨酸 七氟醚 

分 类 号：R686[医药卫生—骨科学]