氯胺酮对大鼠神经元缺氧时线粒体功能的影响

Effect of ketamine on mitochondriai function of rat neurons subjected to anoxia

目的探讨氯胺酮对大鼠神经元缺氧时线粒体功能的影响。方法原代大鼠海马神经元以5×105～1×106/ml的密度接种于35 mm培养皿，采用随机数字表法分为3组（n＝11）：对照组、缺氧组和氯胺酮组。缺氧组于培养液中通入90%N2＋10%CO2 50 ml/min缺氧5 min；氯胺酮组缺氧前1 h，于培养液中加入氯胺酮，终浓度为20 μmol/L，随后处理同缺氧组。各组处理结束后，采用台盼蓝染色，确定神经元死亡率；采用ATP生物发光法测ATP含量；采用罗丹明123染色法测定线粒体膜电位（△Ψm）。结果与对照组比较，缺氧组和氯胺酮组海马神经元死亡率升高，ATP含量和△Ψm降低（P〈0.05）；与缺氧组比较，氯胺酮组海马神经元死亡率降低，ATP含量和△Ψm升高（P〈0.05）。结论氯胺酮减轻大鼠神经元缺氧性损伤的机制与改善线粒体功能有关。

ObjectiveTo investigate the effect of ketamine on the mitochondrial function of rat neurons subjected to anoxia.MethodsPrimarily cultured rat hippocampal neurons were seeded in culture dishes（35 mm in diameter）at the density of 5×105-1×106 cells/ml, and divided into 3 groups（n=11 each）using a random number table： control group, anoxia group and ketamine group.The neurons were exposed to 90% N2 plus 10% CO2 50 ml/min for 5 min in anoxia group.In ketamine group, ketamine was added to the culture medium with the final concentration of 20 μmol/L at 1 h before anoxia, and then the neurons were exposed to 90% N2 plus 10% CO2 50 ml/min for 5 min.After the end of treatment in each group, the dead neurons were detected using trypan blue staining, the ATP content was determined by ATP bioluminescence assay, and mitochondrial membrane potential was measured by rhodamine 123 staining.ResultsCompared with control group, the mortality rate of hippocampal neurons was significantly increased, and the ATP content and mitochondrial membrane potential were significantly decreased in anoxia group and ketamine group（P〈0.05）. Compared with anoxia group, the mortality rate of hippocampal neurons was significantly decreased, and the ATP content and mitochondrial membrane potential were significantly increased in ketamine group（P〈0.05）.ConclusionThe mechanism by which ketamine ameliorates anoxia-induced damage to rat neurons is related to improved mitochondrial function.