不同浓度异丙酚对新生大鼠离体海马神经元凋亡的影响

Effects of different concentrations of propofol on apoptosis in hippocampal neurons in neonatal rats in vitro

目的评价不同浓度异丙酚对新生大鼠离体海马神经元凋亡的影响。方法原代SD新生大鼠海马神经元体外培养7d，采用随机数字表法，将其随机分为7组（n=8）：对照组（C组）正常培养；不同浓度异丙酚组（P1-3组）分别用含异丙酚4、8、12μg／ml的培养基换液处理，继续培养；不同浓度脂肪乳组（F。组）分别用含与P。组药物等浓度、等容量的脂肪乳培养基换液处理，继续培养。孵育24h时倒置相差显微镜下观察细胞形态学结果，免疫组化法检测半胱氨酸蛋白酶-3（caspase-3）蛋白的表达，双染法检测神经元的凋亡。结果与C组比较，P1-3组神经元凋亡率及caspase-3蛋白表达水平升高（P〈0．05或0．01）；P1-3组神经元凋亡率及caspase-3蛋白表达水平依次升高（P〈0．05）；与P1-3组比较，F1-3组神经元凋亡率及caspase-3蛋白表达水平降低（P〈0．05）；F1-3组神经元凋亡率及caspase-3蛋白表达差异无统计学意义（P〉0．05）。P1-3组神经元发生损伤，以B组最明显。结论异丙酚呵促进新生大鼠海马神经元的凋亡，且呈浓度依赖性。

Objective To investigate the effects of different concentrations of propofol on the apoptosis in hippocampal neurons in neonatal rats in vitro. Methods Primary hippocampal neurons were prepared from new- born Sprague-Dawley rats （aged 7 days） and cultured for 7 days. The neurons were randomly divided into 7 groups （ n = 8 each） ： control group （group C） , propofol 4, 8 and 12 μg/ml groups （groups P1-3 ） , and fat emulsion 4, 8 and 12 μg/ml groups （groups F1-3 ） . The cells were cultured for 24 h in the culture medium containing propofol 4, 8 and 12 μg/ml in groups P1-3 , respectively. The cells were cultured for 24 h in the culture medium containing fat emulsion 4, 8 and 12 μg/ml in groups F1-3 , respectively. The cell morphology was examined by microscopy after 24 h culture. The expression of caspase-3 （by immuno-histochemistry） and neuronal apoptosis were detected. The neuronal apoptosis rate was calculated. Results Compared with group C, the neuronal apoptosis rate and caspase- 3 expression were significantly increased in groups Pi-3 （ P 〈 0.05 or 0.01 ） . The neuronal apoptosis rat and caspase-3 expression were increased in a concentration-dependent manner in groups P1-3 （ P 〈 0.05） . The neuronal apoptosis rate and caspase-3 expression were significantly lower in groups FI_3 than in groups P1-3 （ P 〈 0.05 ） . There was no significant difference in the neuronal apoptosis rate and easpase-3 expression between groups F1-3 （ P 〉 0.05 ）. The damage to neurons was induced in groups P1-3 and most severe in group P3 .Conclusion Propofol can promote the apoptosis in hippoeampal neurons in neonatal rats in vitro in a concentration-dependent manner.