七氟烷诱导的miR-211-5p对新生小鼠发育期神经毒性的影响及可能机制
Effects and possible mechanisms of sevoflurane-induced miR-211-5p on developmental neurotoxicity in neonatal mice
高婷婷 1张晓敏 1杜薇 1黄泽清1
作者信息
- 1. 中国医科大学肿瘤医院辽宁省肿瘤医院麻醉科,辽宁沈阳 110042
- 折叠
摘要
目的 探究七氟烷诱导的miR-211-5p对新生小鼠发育期神经毒性的影响及可能机制.方法 将出生后第6天的新生小鼠随机分为对照组、七氟烷组、七氟烷+antagomiR-NC组和七氟烷+antagomiR-211-5p组,每组6只.RT-qPCR检测新生小鼠海马组织miR-211-5p表达水平;HE染色检测新生小鼠海马组织病理损伤;TUNEL实验检测新生小鼠海马组织神经细胞凋亡;RT-qPCR检测新生小鼠海马组织IL-6和TNF-α mRNA表达;生物信息学预测miR-211-5p与NQO1 mRNA的潜在结合位点,双荧光素酶报告基因实验验证;Western blot检测新生小鼠海马组织NQO1和PI3K/AKT/mTOR信号通路相关蛋白表达.结果 七氟烷上调新生小鼠海马组织miR-211-5p表达,促进海马组织病理损伤、神经细胞凋亡和炎症因子表达水平;antagomiR-21 1-5p降低七氟烷诱导的新生小鼠海马组织miR-211-5p表达,抑制海马组织病理损伤、神经细胞凋亡和炎症因子表达水平.miR-211-5p与NQO1 mR-NA靶向结合.七氟烷降低新生小鼠海马组织NQO1蛋白表达,抑制PI3K/AKT/mTOR信号通路激活;antagomiR-211-5p增加七氟烷处理的新生小鼠海马组织NQO1蛋白表达,激活PI3K/AKT/mTOR信号通路.结论 七氟烷通过诱导miR-211-5p下调NQO1抑制PI3K/AKT/mTOR信号通路导致发育期神经毒性.
Abstract
Objective To investigate the effect of sevoflurane-induced miR-211-5p on developmental neurotoxicity in neonatal mice and its possible mechanism.Methods Neonatal mice on the 6th day after birth were randomly divided into control group,sevoflurane group,sevoflurane+antagomiR-NC group and sevoflurane+antagomiR-211-5p group,with 6 mice in each group.RT-qPCR was used to detect the expression level of miR-211-5p in hippocampus of neonatal mice;HE staining was used to detect the pathological damage of hippocampus of neonatal mice;TUNEL assay was used to detect the apoptosis of nerve cells in hippocampus of neonatal mice;RT-qPCR was used to detect the mRNA expression of IL-6 and TNF-α in hippocampus of neonatal mice;bioinformatics was used to predict the potential binding sites of miR-211-5p and NQO1 mRNA,and double luciferase reporter gene assay was used to verify the prediction;Western blot was used to detect the expressions of NQO1 and PI3K/AKT/mTOR signaling pathway related proteins in hippocampus of neonatal mice.Results Sevoflurane upregulated the expression of miR-211-5p in hippocampus of neonatal mice,promoted the pathological damage,nerve cell apoptosis and inflammatory factor expression levels in hippocampus of neonatal mice;antagomiR-211-5p decreased the expression of miR-211-5p in hippocampus of neonatal mice induced by sevoflurane,inhibited the pathological damage,nerve cell apoptosis and inflammatory factor expression levels in hippocampus of neonatal mice.miR-211-5p targeted NQO1 mRNA.Sevoflurane decreased the expression of NQO1 protein in hippocampus of neonatal mice,inhibited the activation of PI3K/AKT/mTOR signaling pathway;antagomiR-211-5p increased the expression of NQO1 protein in hippocampus of neonatal mice treated with sevoflurane,activated PI3K/AKT/mTOR.Conclusion Sevoflurane induces developmental neurotoxicity by down-regulating NQO1 by miR-211-5p and inhibiting PI3K/AKT/mTOR signaling pathway.
关键词
七氟烷/miR-21/1-5p/NQO1/发育期神经毒性/PI3K/AKT/mTOR通路Key words
sevoflurane/miR-211-5p/NQO1/developmental neurotoxicity/PI3K/AKT/mTOR signaling pathway引用本文复制引用
基金项目
辽宁省自然科学基金(20180550218)
出版年
2024
NETL
NSTL
万方数据