基底外侧杏仁核谷氨酸能神经元参与异氟醚麻醉觉醒的调控  

Glutamatergic neurons in basolateral amygdala involved in the regulation of arousal from isoflurane anesthesia

作　　者：王飒 李慧明 王丹 WANG Sa;LI Huiming;WANG Dan(Department of Anesthesiology and Perioperative Medicine,Xijing Hospital,Air Force Medical University,Xi'an 710032,China)

机构地区：[1]空军军医大学西京医院麻醉与围术期医学科,陕西西安710032

出　　处：《空军军医大学学报》2024年第4期369-374,共6页Journal of Air Force Medical University

基　　金：国家自然科学基金青年科学基金(82101343)。

摘　　要：目的探讨基底外侧杏仁核(BLA)谷氨酸能神经元在异氟醚麻醉中的调控作用。方法采用健康雄性8~12周龄Vglut2-Cre转基因小鼠。15只小鼠随机分为光遗传激活组(ChR2组)、抑制组(GtACR组)和对照组(mCherry组),每组5只,分别在BLA核团立体定位注射兴奋性光遗传病毒rAAV2/9-EF1α-DIO-hChR2-mCherry-WPRE-hGH pA、抑制性光遗传病毒rAAV2/9-EF1α-DIO-GtACR-mCherry-WPRE-hGH pA或对照病毒rAAV2/9-EF1α-mCherry-WPRE-hGH pA,并埋置陶瓷插芯。待病毒表达3周后,对待试小鼠进行14 mL/L异氟醚麻醉,并全程记录小鼠皮层脑电(EEG),爆发抑制比(BSR)稳定时光遗传调控BLA谷氨酸神经元,比较光刺激前2 min与刺激2 min时的BSR。21只Vglut2-Gre小鼠随机分为化学遗传激活组(hM3Dq组)、抑制组(hM4Di组)和对照组(mCherry组),每组7只,分别在双侧BLA核团注射化学遗传病毒,3周后进行化学遗传调控实验,比较稳定深度麻醉状态中同一时间点各组EEG频谱及各频段百分比总功率以及翻正反射消失及恢复(LORR/RORR)时间。结果与光刺激前相比,光刺激期间ChR2组BSR降低(P<0.05),GtACR组BSR增加(P<0.05),而mCherry组BSR无统计学差异(P>0.05)。化学遗传实验中,与mCherry组相比,深度麻醉状态下,hM4Di组皮层EEG的α、β、γ频段百分比总功率明显增加(P<0.05),hM3Dq组EEG的α、β、γ频段百分比总功率无统计学差异(P>0.05);hM3Dq组和hM4Di组LORR的时间无统计学差异(P>0.05),而hM3Dq组RORR的时间缩短,hM4Di组RORR的时间延长(P<0.01)。结论BLA谷氨酸神经元参与调控小鼠异氟醚麻醉的觉醒过程,激活BLA谷氨酸神经元,可以显著促进小鼠从异氟醚麻醉状态中觉醒。Objective To investigate the role of glutamatergic neurons in the basolateral amygdala(BLA)in regulating isoflurane anesthesia.Methods Healthy male Vglut2-Cre transgenic mice aged 812 weeks were used in the study.Fifteen mice were randomly divided into optogenetic activation group(ChR2 group),optogenetic inhibition group(GtACR group)and optogenetic control group(mCherry group),with 5 mice in each group.Excitatory optogenetic virus rAAV2/9-EF1α-DIO-hChR2-mCherry-WPRE-hGH pA,inhibitory optogenetic virus rAAV2/9-EF1α-DIO-GtACR-mCherry-WPRE-hGH pA,and controlled virus rAAV2/9-EF1α-mCherry-WPRE-hGH pA were stereotaxically injected into BLA with embedded ceramic ferrules.After virus expression for 3 weeks,the test mice were anesthetized with 14 mL/L isoflurane,the cortical electroencephalography(EEG)of the mice was recorded,BLA glutamatergic neurons were optogenetically regulated during the stable time of burst-suppression ratio(BSR),and BSR was compared 2 min before and 2 min after light stimulation.Twenty-one Vglut2-Cre mice were randomly divided into chemogenetic activation group(hM3Dq group),chemogenetic inhibition group(hM4Di group)and chemogenetic control group(mCherry group),with 7 mice in each group.Chemogenetic regulation experiment was carried out 3 weeks after the chemogenetic virus was injected into bilateral BLA nuclei.The EEG spectrum and the percentage of total power of each frequency band were compared in each group at the same time point in stable and deep anesthesia,as well as the time to loss of righting reflex and return of righting reflex(LORR/RORR).Results Compared with that before light stimulation,BSR in ChR2 group decreased(P<0.05)and BSR in GtACR group increased(P<0.05)during light stimulation,while there was no statistical difference in mCherry group(P>0.05).In the chemogenetic experiment,compared with mCherry group,the percentage of total power ofα,βandγband of cortical EEG in hM4Di group was significantly increased under deep anesthesia(P<0.05),while that in hM3Dq group was not signif

关 键 词：基底外侧杏仁核 谷氨酸能神经元 异氟醚 觉醒 遗传学技术 

分 类 号：R614[医药卫生—麻醉学]