不同浓度异氟烷对大鼠原代神经元细胞毒性作用

Effect of different concentrations of isoflurane on the apoptosis of developing neurons

目的探讨异氟烷对体外原代培养新生鼠大脑神经元细胞的影响。方法出生48 h内清洁级雌雄不限SD大鼠,当天取大脑行皮质神经细胞原代培养,种植后置于温箱中培养。于培养第7天,建立体外培养大脑神经元暴露异氟烷模型,将同一批体外培养的神经元随机分为6组:正常温箱空白对照组(C组)、吸入0.5、1.0、1.5、2.0和2.5 MAC浓度的异氟烷实验组(P1、P2、P3、P4、P5组)。C组置于温箱中不行任何处理;通入21%O2,5%CO24 h后立即采用噻唑蓝(MTT)法分别测定6组神经元的细胞活力,细胞凋亡数。实验共重复6次。结果 P1、P2和P3组皮质神经元存活率和凋亡率与C组比较差异无统计学意义(P>0.05);P4和P5组皮质神经元存活率降低,细胞凋亡数增加(P<0.05)。结论较高浓度异氟烷(2.0和2.5 MAC)会降低原代神经元生存率,增加细胞凋亡,产生神经毒性作用。

Objective To investigate the effect of isoflurane on the cytotoxicity of primary cultured cortical neurons.Methods Postnatal day 2 Sprague-Dawley rats were used for primary cortical culture. Cells were seeded in 96-well plates and then placed in an incubator. After 7 days, the cultured cortical neurons were randomly divided into 6 groups ：normal control in the incubator（group C）; Isoflurane group which seated in the sealed plastic box with 21%O2, 5%CO2,and four different concentrations of isoflurane delivered from an anesthesia machine for 4 h（group P1 ：0.5MAC isoflurane; group P2 ：1 MAC isoflurane; group P3 ：1.5MAC isoflurane; group P4 ：2 MAC isoflurane; group P5 ： 2.5MAC isoflurane）. Cytotoxicity was evaluated by MTT reduction assays. Results There were no significant difference in the rate of survival and apoptosis in neurons between group P1,P2,P3 and group C. The rate of survival and apoptosis in neurons in both group P4 and P5 were significant higher than that in group C（P〈0.01）. Conclusion Higher concentrations of isoflurane（2 and 2.5 MAC） for 4 h caused significant cytotoxicity of primary cultured cortical neurons.