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目的研究氯化锂(LiCl)对中枢神经系统发育期七氟醚神经毒性的影响,以及β-arrestin2/AKT信号通路在其中所起的作用。方法对离体培养7 d的海马神经细胞及出生后7 d的Sprague-Dawley幼鼠进行七氟醚处理(3%,6 h),制备七氟醚神经毒性模型。离体实验中给予Li Cl或Li Cl+β-arrestin2 si RNA转染处理后,应用流式细胞仪、MTT及Hoechst染色检测细胞凋亡;应用免疫印迹检测total AKT、Phospho-AKT、total GSK3β、Phospho-GSK3β、Bax及Bcl-2的蛋白表达水平;应用条件恐惧实验及Morris水迷宫检测实验动物的情绪及空间学习记忆功能变化。结果七氟醚处理(3%,6 h)使海马神经细胞凋亡增加,应用Li Cl则可明显缓解七氟醚神经毒性。七氟醚处理可降低Phospho-AKT及Bcl-2的蛋白表达,增加Phospho-GSK3β及Bax的蛋白表达;LiCl可增加Phospho-AKT及Bcl-2表达,降低Phospho-GSK3β及Bax表达水平。Li Cl的神经保护作用可被转染β-arrestin2 si RNA、SH-5或LY294002所逆转。Li Cl明显缓解了中枢神经系统发育期七氟醚处理引起的情绪及空间学习记忆功能异常。结论 Li Cl可缓解中枢神经系统七氟醚神经毒性,其机制可能与β-arrestin2依赖的AKT/GSK3β信号通路有关。
Objective To investigate the effects of LiCl on the neurotoxicity of sevoflurane in CNS development and the role of β-arrestin2 / AKT signaling pathway in CNS development. Methods Sevoflurane neurotoxicity model was established by sevoflurane treatment (3%, 6 h) in cultured hippocampal neurons cultured in vitro and Sprague-Dawley rats 7 days after birth. After transfection with LiCl or LiCl + β-arrestin2 si RNA in vitro, apoptosis was detected by flow cytometry, MTT and Hoechst staining. Total AKT, Phospho-AKT, total GSK3β, Phospho-GSK3β, Bax and Bcl-2 protein expression levels; conditions of fear experiments and Morris water maze test animals emotional and spatial learning and memory function changes. Results Sevoflurane treatment (3%, 6 h) increased the apoptosis of hippocampal neurons, and the application of Li Cl significantly attenuated the neurotoxicity of sevoflurane. Sevoflurane could decrease the protein expression of Phospho-AKT and Bcl-2 and increase the protein expression of Phospho-GSK3β and Bax. LiCl could increase the expression of Phospho-AKT and Bcl-2 and decrease the expression of Phospho-GSK3β and Bax. The neuroprotective effect of LiCl can be reversed by transfection of beta-arrestin2 si RNA, SH-5 or LY294002. LiCl significantly relieved sevoflurane-induced sentiment and spatial learning and memory dysfunction in central nervous system development. Conclusion Li Cl can relieve the neurotoxicity of sevoflurane in the central nervous system, which may be related to the β-arrestin2-dependent AKT / GSK3β signaling pathway.