七氟烷对大鼠原代培养海马神经元细胞毒性作用

Effect of sevoflurane on cytotoxicity of primary cultured hippocampal neurons of rats

目的探讨吸入麻醉药物七氟烷对原代培养的SD乳鼠大脑海马神经元的细胞毒性作用。方法新生1d的SD乳鼠，雌雄不限，当天取脑行大脑海马细胞原代培养，种植于96孔板（15孔／板）后置于温箱中培养，于培养第7天，建立体外培养大脑海马神经元暴露七氟烷模型，将同一批体外培养的海马神经元随机分为3组：正常温箱空白对照组（C1）、实验对照组（C2）、吸入麻醉药物七氟烷组（S组），C1组置于温箱中不行任何处理；C2组和S组置于相同条件的密闭容器中（5％二氧化碳，21％氧气，37℃），s组同时给予3％七氟烷4h，4h后立即采用噻唑蓝（MTT）法分别测定3组神经元的细胞活力。实验共重复3次。结果与C1组和C2组比较，S组MTT值下降，与前两组比较差异有统计学意义（P〈0．05）；C1组和C2组之间差异无统计学意义（P〉0．05）。结论本实验成功建立了体外培养海马神经元暴露七氟烷模型，模型中七氟烷直接作用于纯度较高的海马神经元，且模型中除七氟烷外的其他环境条件不会对培养的大脑海马神经元细胞活力产生影响；体外给予3％吸入麻醉药物七氟烷4h会降低大脑海马神经元的细胞活力，产生神经毒性作用。

Objective To investigate the effect of sevofluranc on the cytotoxicity of primary cultured hippocampal neurons. Methods Postnatal day 1 Spraguc-Dawley rats were used for primary hippocampal culture. Cells were seeded in 96-well plates and then placed in an incubator. After 7 days, the cultured hippocampal neurons were randomly divided into 3 groups （n = 10 each） ： group I normal control in the incubator （ group C1 ） ; group II another control group which seated in the sealed plastic box with 21% 02, 5% CO2 delivered from an anesthesia machine for 4 h （group C2） ; group III sevoflurane group which seated in the sealed plastic box with 21% O2 , 5% CO2 , and 3% sevoflurane delivered from an anesthesia machine for 4 h （ group S）. Then cellular effects of sevoflurane and other conditionings were evaluated by 3-（4, 5-dimethyithiazol-2-yl）-2, 5-diphenyl-tetra-zolium bromide （MTT） reduction assays. Results There was no significant difference between group C1 and C2 （ P 〉 0. 05）. In group S, conditioning with sevoflurane for a total of 4 h resulted in a decrease in MTT as compared with group C1 and C2 （P 〈 0. 05 ）. Conclusion In this experiment, We successfully made a model for cultured ceils to inhale sevoflurane in which experimental conditions inside tight gas container itself had no effect on cell viability determined by MTT reduction assays. Our resuhs suggest 3% sevoflurane for 4 h caused significant cytotoxicity, with reduction of MTT.