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 anesthe-sia-emergence in mice
郭晓雨 1吴慧敏 2王丹 3董海龙4
作者信息
- 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;榆林市第一医院麻醉科,榆林 719000
- 2. 陕西中医药大学第二附属医院中心实验室,咸阳 712046
- 3. 空军军医大学西京医院麻醉与围术期医学科,西安 710032
- 4. 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;空军军医大学西京医院麻醉与围术期医学科,西安 710032
- 折叠
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.
Abstract
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 de-termination of the expression of c-Fos and the rate of co-labeling with GABA neurons.For the optogenetic ex-periment,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-Efla-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 oxy-gen group,the rate of c-Fos co-labeling with GABA neurons in the BNST brain region was significantly re-duced(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 region may be involved in the process of loss of consciousness in isoflurane-anesthetized mice,while increased viability of GABAergic neurons in the BNST brain region promotes the transition from anesthesia to emergence.
关键词
异氟醚/麻醉,全身/GABA能神经元/终纹床核Key words
Isoflurane/Anesthesia,general/GABAergic neurons/Bed nucleus of the stria terminalis引用本文复制引用
基金项目
国家自然科学基金青年基金(82101343)
出版年
2024
NETL
NSTL
万方数据