国际麻醉学与复苏杂志   2014, Issue (4): 2-2
    
不同浓度异丙酚对胎鼠海马神经元存活及凋亡的影响
徐晓东, 吴国华, 张良成1()
1.福建医科大学附属协和医院
Effect of different concentration of propofol on cell viability and apoptosis in hippocampal neurons of fetal rats
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摘要：

目的 探讨不同浓度异丙酚对胎鼠离体海马神经元存活及凋亡的影响。　方法　SD大鼠妊娠17 d~18 d后取胎鼠海马神经元体外培养7 d，应用免疫细胞化学法鉴定神经元并作为研究对象。① 采用随机数字表法将其随机分为12组（每组设5个平行副孔，实验重复5次）：对照组（C组）正常培养，异丙酚溶剂对照组（D组）用含0.1%二甲基亚砜（dimethyl sulfoxide， DMSO）的培养基处理，不同浓度异丙酚组（P1~10组）分别用含异丙酚终浓度为0.01、0.1、0.5、1、5、10、50、100、150、200 μmol/L的培养基换液处理，继续培养24 h后用3-（4，5-二甲基噻唑-2）-2，5-二苯基四氮唑溴盐［3-（4，5-Dimethylthiazol-2-yl）-2，5-diphenyltetrazolium bromide， MTT］法检测细胞生存率；② 采用随机数字表法将其随机分为7组（实验重复5次）：对照组（C组）正常培养，异丙酚溶剂对照组（D组）用含0.1%DMSO的培养基处理，不同浓度异丙酚组（PI、II、III、IV、V组）分别用含异丙酚终浓度为0.1、1、10、100、200 μmol/L的培养基换液处理，继续培养24 h，免疫荧光细胞化学染色观察细胞核形态学结果，Annexin-V-FLUOS双染流式细胞术检测海马神经元凋亡率。　结果　① 与C组比较，D组海马神经元生存率差异无统计学意义（P>0.05）；与D组比较，P1、2、5组生存率差异无统计学意义（P>0.05），P3组（102.2±2.7）%、P4组（102.9±2.7）%生存率升高（P<0.05），而P6~10组生存率分别为（97.0±3.5）%、（90.1±1.7）%、（79.8±0.9）%、（67.9±1.1）%、（42.3±2.2）%，呈浓度依赖性降低（P<0.05或P<0.01）；② 与C组比较，D组海马神经元凋亡率差异无统计学意义（P>0.05）；与D组比较，PI组凋亡率差异无统计学意义（P>0.05），PII组（4.7±0.7）%凋亡率明显降低（P<0.05），而PIII、IV、V组凋亡率分别为（8.2±0.8）%、（18.1±1.1）%、（29.1±2.1）%，呈浓度依赖性升高（P<0.05或P<0.01）。　结论　异丙酚对胎鼠离体海马神经元生存及凋亡的影响呈双向作用，1 μmol/L浓度异丙酚能够对抗海马神经元凋亡，提高细胞存活率，而10、100、200 μmol/L浓度的异丙酚则呈剂量依赖性地促进海马神经元凋亡，降低细胞生存率。
关键词： 二异丙酚；海马神经元；细胞生存率；凋亡
Abstract:

Objective To investigate the effect of different concentration of propofol on cell viability in hippocampal neurons of fetal rats in vitro.　Methods　Primary cultured hippocampal neurons were prepared from the fetuses of Sprague-Dawley rats after 17 d-18 d of gestation， and cultured for 7 d， immunocytochemical method was used to identify the cultured neurons. ① The neurons were randomly divided into 12 groups（n=5）： control group （group C） received no treatment，in group D 0.1% dimethyl sulfoxide（DMSO） was added，in group P1-10 propofol was added at the final concentration of 0.01， 0.1， 0.5， 1， 5， 10， 50， 100， 150 μmol/L and 200 μmol/L respectively． The cells were selected after 24 h incubation for detection of the cell survival rate by cell viability［3-（4，5-Dimethylthiazol-2-yl）-2，5-diphenyltetrazolium bromide （MTT） assay］. ② The neurons were randomly divided into 7 groups（n=5）：control group（group C） received no treatment，in group D 0.1% DMSO was added，in group PI，II，III，IV，V propofol was added at the final concentration of 0.1， 1， 10， 100 μmol/L and 200 μmol/L respectively. The cells were selected after 24 h incubation for detection of the neuronal apoptosis by immunofluorescence staining and flow cytometry.　Results　① There was no significantly difference in the cell survival rate between group C and D（P>0.05）. Compared with group D， the cell survival rate was no significantly difference in group P1，2，5（P>0.05），the cell survival rate was significantly increased in group P3（102.2±2.7）%，P4（102.9±2.7）%（P<0.05） and that was significantly lower in group P6-10 in a concentration-dependent manner by（97.0±3.5）%，（90.1±1.7）%，（79.8±0.9）%，（67.9±1.1）%，（42.3±2.2）% respectively（P<0.05 or P<0.01）. ② There was no significantly difference in the neuronal apoptosis rate between group C and D（P>0.05）. Compared with group D， the neuronal apoptosis rate was no significantly difference in group PI（P>0.05），the neuronal apoptosis rate was significantly decreased in group PII（P<0.05） and that was significantly higher in group PIII，IV，V in a concentration-dependent manner by （8.2±0.8）%， （18.1±1.1）%， （29.1±2.1）% respectively（P<0.05 or P<0.01）.　Conclusions　The effect of propofol on cell viability and apoptosis in hippocampal neurons was bidirectional，propofol at the concentration of 1 μmol/L can inhibit the apoptosis and improve the survival rate of neurons，while decrease the cell viability through promoting the cell apoptosis in a concentration-dependent manner at the concentration of 10，100 μmol/L and 200 μmol/L．
Key words: Propofol；Hippocampal neuron；Cell viability；Apoptosis