异氟烷对小鼠神经干细胞BDNF/Caspase3及Notch信号相关基因表达的影响

Effect of Isoflurane on Expression of BDNF/Caspase3 and Notch in Cultured Neural Stem Cells

目的:探讨异氟烷对小鼠神经干细胞的BDNF、Caspase3及Notch信号相关基因表达的影响。方法:给予体外培养的新生小鼠海马神经干细胞不同浓度异氟烷处理,实验分为对照组和异氟烷处理组(ISO1.0,ISO1.5),其中异氟烷组细胞分别给予1.0MAC和1.5 MAC两个浓度的异氟烷处理2小时,对照组给予O_2处理2小时,随后置于培养箱正常培养24小时后收集细胞,提取细胞RNA检测BDNF,Caspase3及Notch相关基因(Notch2、Notch 3和Hes5)的m RNA水平变化。结果:与对照组相比,(1)异氟烷组小鼠神经干细胞的功能基因BDNF m RNA水平下调,凋亡相关基因Caspase3的m RNA水平上调;(2)异氟烷组神经干细胞的Notch2和Notch3受体m RNA表达下调,Notch信号通路靶基因Hes5的m RNA水平也明显下调;(3)异氟烷对神经干细胞的作用具有剂量依赖性,浓度越高对神经干细胞BDNF、Caspase3及Notch信号相关基因表达的影响越大。结论:异氟烷可能通过抑制小鼠神经干细胞的Notch信号通路,下调BDNF的m RNA表达,上调Caspase3的m RNA水平,影响神经干细胞的正常功能。

Objective： To explore the effect of isoflurane on the expression of brain derived neurotrophic factor（BDNF）, Caspase3 and Notch signaling in neural stem cells. Methods： Cultured hippocampal neural stem cells（NSCs） from newborn mice were divided into control and isoflurane groups： control group was treated with O_2 for 2 hours and isoflurane treatment groups（ISO1.0 or ISO1.5） were treated with 1.0 MAC and 1.5 MAC of isoflurane for 2 hours respectively. Then cells were cultured in the incubator with 5% CO_2and37℃ for 24 hours. And the total RNA was extracted from cells and the m RNA levels of BDNF, Caspase3, Notch2, Notch3, and Hes5 were measured by Real-time PCR. Results：（1） Compared with the control, isoflurane significantly down-regulated the BDNF m RNA level and up-regulated the Caspase3 m RNA level in the neural stem cells;（2） Isoflurane significantly decreased the m RNA levels of Notch2, Notch3 and Hes5 in the neural stem cells;（3） The effect of isoflurane revealed a dose effect---compared with ISO1.0 group, cells treated with 1.5 MAC of isoflurane suffered more influence. Conclusion： Isoflurane exposure suppressed the BDNF expression and increased apoptosis of cultured neural stem cells, and this effect possibly mediated by Notch signal pathway.