异氟烷对新生小鼠神经元细胞外信号调节激酶和cAMP反应元件结合蛋白磷酸化水平的影响

Effect of Isoflurane exposure on expressions of ERKs and CREB in neuron of newborn mice

目的观察异氟烷对小鼠神经元细胞外信号调节激酶和环磷酸腺苷(c AMP)反应元件结合蛋白磷酸化水平的影响,探究异氟烷诱导神经元损伤和学习记忆障碍的可能机制。方法原代培养小鼠神经元,分为对照组、0.96 mmol/L异氟烷处理12 h及24 h组。采用MTS方法检测各组细胞活力;Western blot方法检测p-ERK1/2和p-CREB表达情况;逆转录定量PCR(RT-q PCR)方法检测ERK和CREB m RNA表达。结果与对照组比较,0.96 mmol/L异氟烷处理12 h及24 h后神经元活力明显下降(P<0.05);Western blot实验结果显示:与对照组比较,异氟烷处理组小鼠神经元中p-ERK1/2和p-CREB表达水平均明显降低(P<0.05);随着异氟烷处理时间的增加,p-ERK1/2和p-CREB表达逐渐降低,处理12 h与24 h比较差异有统计学意义(P<0.05)。RT-q PCR实验显示:与对照组比较,异氟烷处理组小鼠神经元中ERK和CREB m RNA表达水平比较,差异无统计学意义(P>0.05)。结论一定浓度的异氟烷可以降低原代培养的小鼠神经元活力和ERK1/2、CREB的磷酸化水平。

Objective To investigate Isoflurane-induced neuronal damage and possible mechanisms involved in learning and memory impairments. Methods Cerebral neurons separated from fetal mice were cultured and divided into three groups, one control group, two experimental groups were treated with 0.96 mmol/L Isoflurane for 12 h and 24 h, respectively. MTS method was used to determine the cell viability; Western blot method was used to detect p-ERK1/2 and p-CREB expressions; the reverse transcription qPCR （RT-qPCR） method was used to detect mRNA expressions of ERK and CREB. Results Compared with the control group, the viability of neuronal cells decreased significantly （P 〈 0.05）; the phosphorylation of ERK1/2 and CREB also significantly decreased （P 〈 0.05）, and with the prolonging of Isoflurane treatment duration, the expression of p-ERK1/2 and p-CREB decreased gradually, and the difference between 12 and 24 h after treatment ＇was statistically significant （P 〈 0.05）; there were no statistically significant difference in mRNA expressions of the ERK and CREB. （P 〉 0.05）. Conclusion A certain concentration of Isoflurane may directly inhibit neuronal cell viability and also inhibits the activities of ERK1/2 and CREB.