CAJ | 학술논문

目的探讨吸人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.
목적 탐토흡인마취약물칠불완대원대배양적SD유서대뇌해마신경원적세포독성작용.방법 신생1d적SD유서,자웅불한,당천취뇌행대뇌해마세포원대배양,충식우96공판(15공/판)후치우온상중배양,우배양제7천,건입체외배양대뇌해마신경원폭로칠불완모형,장동일비체외배양적해마신경원수궤분위3조:정상온상공백대조조(C1)、실험대조조(C2)、흡입마취약물칠불완조(S조),C1조치우온상중불행임하처리；C2조화S조치우상동조건적밀폐용기중(5％이양화탄,21％양기,37℃),S조동시급여3％칠불완4h,4h후립즉채용새서람(MTT)법분별측정3조신경원적세포활력.실험공중복3차.결과 여C1조화C2조비교,S조MTT치하강,여전량조비교차이유통계학의의(P＜0.05)；C1조화C2조지간차이무통계학의의(P＞0.05).결론 본실험성공건립료체외배양해마신경원폭로칠불완모형,모형중칠불완직접작용우순도교고적해마신경원,차모형중제칠불완외적기타배경조건불회대배양적대뇌해마신경원세포활력산생영향；체외급여3％흡입마취약물칠불완4h회강저대뇌해마신경원적세포활력,산생신경독성작용.
Objective To investigate the effect of sevoflurane on the cytotoxicity of primary cultured hippocampal neurons.Methods Postnatal day 1 Sprague-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％ O2,5％ 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 cells to inhale sevoflurane in which experimental conditions inside tight gas container itself had no effect on cell viability determined by MTT reduction assays.Our results suggest 3％ sevoflurane for 4 h caused significant cytotoxicity,with reduction of MTT.