终纹床核GABA能神经元在小鼠异氟烷全麻-觉醒中的作用  

Role of GABAergic neuron in bed nucleus of stria terminalis in isoflurane-induced general anesthesia-emergence in mice

作　　者：郭晓雨 吴慧敏 王丹 董海龙 Guo Xiaoyu;Wu Huimin;Wang Dan;Dong Hailong(Jiangsu Province Key Laboratory of Anesthesiology Xuzhou Medical University,Jiangsu Province Key Laboratory of Anesthesia and Analgesia Application Technology NMPA Key Laboratory for Research and Evaluation of Narcotic and Psychotropic Drugs,Xuzhou 221004,China;Department of Anesthesiology,The First Hospital of Yulin,Yulin 719000,China;Central Laboratory,Second Affiliated Hospital of Shaanxi University of Chinese Medicine,Xianyang 712046,China;Department of Anesthesiology and Perioperative Medicine,Xijing Hospital,The Fourth Military Medical University,Xi'an 710032,China)

机构地区：[1]徐州医科大学江苏省麻醉学重点实验室、徐州医科大学江苏省麻醉与镇痛应用技术重点实验室、国家药品监督管理局麻醉精神药物研究与评价重点实验室,徐州221004 [2]榆林市第一医院麻醉科,榆林719000 [3]陕西中医药大学第二附属医院中心实验室,咸阳712046 [4]空军军医大学西京医院麻醉与围术期医学科,西安710032

出　　处：《中华麻醉学杂志》2024年第5期587-592,共6页Chinese Journal of Anesthesiology

基　　金：国家自然科学基金青年项目(82101343)。

摘　　要：目的探讨终纹床核(BNST)GABA能神经元在小鼠异氟烷全麻-觉醒中的作用。方法健康雄性Vgat-Cre转基因小鼠23只,8~10周龄,体质量约22 g。免疫荧光染色实验:取8只小鼠,采用随机数字表法分为2组(n=4):氧气组和异氟烷组。氧气组吸入1.0 L/min氧气2 h,异氟烷组吸入1.4%异氟烷+1.0 L/min氧气2 h,随后处死小鼠,取脑组织,用免疫荧光染色法测定c-Fos阳性表达及其与GABA能神经元的共标率。光遗传学实验:取15只小鼠,采用随机数字表法分为3组(n=5):对照组(CON组)、光遗传兴奋组(CHR2组)和光遗传抑制组(eNpHR组),分别于BNST脑区注射rAAV-Ef1a-DIO-mCherry、rAAV-Ef1a-DIO-CHR2-mCherry和rAAV-Ef1a-DIO-eNpHR3.0-mCherry病毒。待病毒表达3周后,小鼠吸入1.0%异氟烷+1.0 L/min氧气,同时检测皮层脑电,当小鼠进入稳定麻醉状态时,采用光遗传方法调控BNST脑区GABA能神经元活力,记录光刺激前2 min与刺激2 min时皮层脑电爆发性抑制率(BSR)。结果与氧气组相比,异氟烷组BNST脑区c-Fos与GABA神经元共标率降低(P<0.05),c-Fos阳性神经元减少。与CON组或光刺激前相比,CHR2组光刺激时BSR降低(P<0.001),eNpHR组BSR差异无统计学意义(P>0.05)。结论BNST脑区GABA能神经元活力降低可能参与异氟烷麻醉小鼠意识消失过程,BNST脑区GABA能神经元活力升高促进麻醉向觉醒转换。Objective To investigate the role of GABAergic neurons in the bed nucleus of the stria terminalis(BNST)in isoflurane-induced general anesthesia-emergence in mice.Methods Twenty-three healthy male Vgat-Cre transgenic mice,aged 8-10 weeks,weighing 22 g,were used in the study.In the immunofluorescence staining experiment,8 mice were selected and divided into 2 groups(n=4 each)using a random number table method:oxygen group and isoflurane group.Oxygen group inhaled oxygen at a rate of 1.0 L/min for 2 h,while isoflurane group inhaled 1.4%isoflurane+1.0 L/min oxygen for 2 h.The animals were then sacrificed,and brain tissues were removed and subjected to immunofluorescence staining for determination of the expression of c-Fos and the rate of co-labeling with GABA neurons.For the optogenetic experiment,15 mice were divided into 3 groups(n=5 each)using a random number table method:control group(CON group),optogenetic excitation group(CHR2 group)and optogenetic inhibition group(eNpHR group).The rAAV-Ef1a-DIO-mCherry,rAAV-Ef1a-DIO-CHR2-mCherry,and rAAV-Ef1a-DIO-eNpHR3.0-mCherry viruses were injected to the BNST brain region.After 3 weeks of virus expression,the mice were exposed to 1.0% isoflurane+1.0 L/min oxygen,and their cortical EEG was simultaneously monitored.When the mice reached a stable anesthetic state,optogenetic methods were utilized to modulate the viability of GABAergic neurons in the BNST brain region,and the burst suppression ratio(BSR)of the cortical EEG was recorded at 2 min before light stimulation and 2 min of light stimulation.Results Compared with oxygen group,the rate of c-Fos co-labeling with GABA neurons in the BNST brain region was significantly reduced(P<0.05),and the c-Fos-positive neurons were reduced in isoflurane group.Compared with CON group or with the prestimulation level,BSR was significantly decreased in CHR2 group(P<0.001),and no significant change was found in BSR during light stimulation in eNpHR group(P>0.05).Conclusions Decreased viability of GABAergic neurons in the BNST brain regio

关 键 词：异氟醚 麻醉 全身 GABA能神经元 终纹床核 

分 类 号：R614[医药卫生—麻醉学]