激活下丘脑背内侧核区星形胶质细胞可加速七氟醚麻醉小鼠觉醒

doi:10.12122/j.issn.1673-4254.2025.04.10

南方医科大学学报 ›› 2025, Vol. 45 ›› Issue (4): 751-759.doi: 10.12122/j.issn.1673-4254.2025.04.10

• • 上一篇

激活下丘脑背内侧核区星形胶质细胞可加速七氟醚麻醉小鼠觉醒

郭舒婷¹(), 曹福羊¹^,²(), 郭永馨¹, 李言响¹^,³, 郝新宇¹, 张倬宁¹, 周志康¹, 仝黎¹(), 曹江北¹()

^1.解放军总医院第一医学中心麻醉科，北京 100853
^2.解放军总医院第六医学中心麻醉科，北京 100048
^3.解放军陆军第七十一集团军医院麻醉科，江苏徐州 221004

收稿日期:2024-12-27 出版日期:2025-04-20 发布日期:2025-04-28
通讯作者: 仝黎,曹江北 E-mail:gstanes@163.com;caofuyang840723@163.com;tongli301@aliyun.com;caojiangbei@301hospital.com.cn
作者简介:郭舒婷，在读硕士研究生，E-mail: gstanes@163.com
曹福羊，在读博士研究生，主治医师，E-mail: caofuyang840723@163.com
第一联系人：郭舒婷、曹福羊共同为第一作者
基金资助:
国家自然科学基金(82271213)

Activation of astrocytes in the dorsomedial hypothalamus accelerates sevoflurane anesthesia emergence in mice

Shuting GUO¹(), Fuyang CAO¹^,²(), Yongxin GUO¹, Yanxiang LI¹^,³, Xinyu HAO¹, Zhuoning ZHANG¹, Zhikang ZHOU¹, Li TONG¹(), Jiangbei CAO¹()

^1.Department of Anesthesia, First Medical Center of Chinese PLA General Hospital, Beijing 100853, China
^2.Department of Anesthesia, Sixth Medical Center of Chinese PLA General Hospital, Beijing 100048, China
^3.Department of Anesthesia, 71st Group Army Hospital of CPLA Army, Xuzhou 221004, China

Received:2024-12-27 Online:2025-04-20 Published:2025-04-28
Contact: Li TONG, Jiangbei CAO E-mail:gstanes@163.com;caofuyang840723@163.com;tongli301@aliyun.com;caojiangbei@301hospital.com.cn
Supported by:
National Natural Science Foundation of China(82271213)

摘要/Abstract

摘要：

目的探究下丘脑背内侧核（DMH）区星形胶质细胞在七氟烷麻醉觉醒中的调节作用。方法选用42只雄性C57小鼠，随机分为6组（n=7），研究星形胶质细胞在七氟烷麻醉中的活性变化时分为EGFP组和GCaMP6组，通过钙成像技术记录星形胶质细胞的活性；研究光遗传激活DMH区星形胶质细胞对麻醉觉醒的影响时，行为学实验分为EGFP组和ChR2组；光遗传脑电记录实验分为EGFP组和ChR2组。麻醉诱导与觉醒的评判标准以翻正反射的消失与恢复为准，在2.0%七氟烷浓度下记录脑电并分析爆发抑制率（BSR），在1.5%七氟烷浓度下分析脑电功率频谱。此外，采用免疫荧光染色观察GFAP阳性细胞（星形胶质细胞）与病毒蛋白信号的共定位。结果随着七氟烷浓度的增加，DMH区星形胶质细胞的活性逐渐降低。在2.0%七氟烷麻醉中，ChR2组的觉醒时间缩短（P<0.05），光遗传激活DMH区星形胶质细胞后BSR降低（P<0.001）。在1.5%七氟烷麻醉中，光激活后ChR2组小鼠的脑电γ波增加（P<0.001），δ波减少（P<0.01）。结论光遗传激活DMH区星形胶质细胞能够促进七氟烷麻醉后的觉醒，但对麻醉诱导过程无显著调节作用。这一发现为麻醉觉醒机制的研究提供了新的视角，可能为术后快速苏醒及麻醉并发症的干预提供潜在靶点。

关键词: 星形胶质细胞, 下丘脑背内侧核区, 七氟烷, 麻醉觉醒

Abstract:

Objective To investigate the regulatory role of astrocytes in the dorsomedial hypothalamus (DMH) during sevoflurane anesthesia emergence. Methods Forty-two male C57BL/6 mice were randomized into 6 groups (n=7) for assessing astrocyte activation in the dorsomedial hypothalamus (DMH) under sevoflurane anesthesia. Two groups of mice received microinjection of agfaABC1D promoter-driven AAV2 vector into the DMH for GCaMP6 overexpression, and the changes in astrocyte activity during sevoflurane or air inhalation were recorded using calcium imaging. For assessing optogenetic activation of astrocytes, another two groups of mice received microinjection of an optogenetic virus or a control vector into the DMH with optic fiber implantation, and sevoflurane anesthesia emergence was compared using behavioral experiments. In the remaining two groups, electroencephalogram (EEG) recording during sevoflurane anesthesia emergence was conducted after injection of the hChR2-expressing and control vectors. Anesthesia induction and recovery were assessed by observing the righting reflex. EEG data were recorded under 2.0% sevoflurane to calculate the burst suppression ratio (BSR) and under 1.5% sevoflurane for power spectrum analysis. Immunofluorescence staining was performed to visualize the colocalization of GFAP-positive astrocytes with viral protein signals. Results Astrocyte activity in the DMH decreased progressively as sevoflurane concentration increased. During 2.0% sevoflurane anesthesia, the mice injected with the ChR2-expressing virus exhibited a significantly shortened wake-up time (P<0.05), and optogenetic activation of the DMH astrocytes led to a marked reduction in BSR (P<0.001). Under 1.5% sevoflurane anesthesia, optogenetic activation resulted in a significant increase in EEG gamma power and a significant decrease in delta power in ChR2 group (P<0.01). Conclusion Optogenetic activation of DMH astrocytes facilitates sevoflurane anesthesia emergence but does not significantly influence anesthesia induction. These findings offer new insights into the mechanisms underlying anesthesia emergence and may provide a potential target for accelerating postoperative recovery and managing anesthesia-related complications.

Key words: astrocytes, dmh region, sevoflurane, anesthesia emergence

郭舒婷, 曹福羊, 郭永馨, 李言响, 郝新宇, 张倬宁, 周志康, 仝黎, 曹江北. 激活下丘脑背内侧核区星形胶质细胞可加速七氟醚麻醉小鼠觉醒[J]. 南方医科大学学报, 2025, 45(4): 751-759.

Shuting GUO, Fuyang CAO, Yongxin GUO, Yanxiang LI, Xinyu HAO, Zhuoning ZHANG, Zhikang ZHOU, Li TONG, Jiangbei CAO. Activation of astrocytes in the dorsomedial hypothalamus accelerates sevoflurane anesthesia emergence in mice[J]. Journal of Southern Medical University, 2025, 45(4): 751-759.

图/表 8

图1 病毒注射脑图谱及行为学实验示意图

Fig.1 Diagram of virus injection into the brain and schematic diagram of behavioral experiment. A: Schematic diagram of calcium imaging virus injection into the DMH of a mouse. B: Schematic diagram of the calcium signal recording system C: Detailed flow chart of calcium imaging experiment, including the time of sevoflurane on and off, LORR and RORR.

图2 光遗传病毒注射及行为学示意图

Fig.2 Schematic diagram of optogenetic virus injection and behavioral experiments. A: Illustration of a coronal brain slice showing the locations of virus injection and optic fiber implantation. B: Schematic diagram of the equipment used for behavioral observation, optic fiber stimulation, and EEG recording during anesthesia. C: Flowchart of optogenetic modulation of behavioral testing under general anesthesia.

图3 钙成像病毒在DMH区荧光的表达与验证

Fig.3 Fluorescence expression and validation of calcium imaging virus in the DMH region.

图4 DMH星形胶质细胞在七氟烷诱导觉醒过程中的钙活性变化

Fig.4 Changes of calcium activity in DMH astrocytes of rats during sevoflurane anesthesia emergence. A: Dynamic monitoring of changes in astrocyte activity during sevoflurane anesthesia. B: Changes in EEG activity during sevoflurane anesthesia corresponding to Fig.A. C: Changes of ΔF/F value of astrocytes in the DMH region during anesthesia induction and awakening. ***P<0.001 vs EGFP group.

图5 免疫荧光染色显示光遗传病毒在小鼠脑组织中的表达情况

Fig.5 Immunofluorescence staining showing the expression of optogenetic virus in mouse brain tissue.

图6 光遗传激活DMH星形胶质细胞的LORR时间和RORR时间

Fig.6 Statistical analysis of LORR time (A) and RORR time (B) of optogenetically activated DMH astrocytes. *P<0.05 vs EGFP group.

图7 在2.0%七氟醚麻醉下光遗传学激活DMH星形胶质细胞的脑电变化

Fig.7 EEG of rats with optogenetically activated DMH astrocytes under 2.0% sevoflurane anesthesia. A: Comparison of representative EEG spectra between EGFP group and ChR2 group under 2.0% sevoflurane anesthesia. B: Comparison of EEG burst suppression rate between EGFP group and ChR2 group after 2 min of light stimulation at 25th min of sevoflurane anesthesia. C: Changes in the brain spontaneous rhythm (BSR%) curve per minute before and after light stimulation. *P<0.05, ***P<0.001 vs EGFP group.

图8 在1.5%七氟醚麻醉下光遗传学激活DMH星形胶质细胞的脑电

Fig.8 EEG of rats with optogenetically activated DMH astrocytes under 1.5% sevoflurane anesthesia. A: Comparison of representative EEG spectra between EGFP group and ChR2 group under 1.5% sevoflurane anesthesia. B: Quantification of EEG power percentage during optogenetic activation in EGFP and ChR2 mice. *P<0.05, **P<0.01, ***P<0.001.

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激活下丘脑背内侧核区星形胶质细胞可加速七氟醚麻醉小鼠觉醒

Activation of astrocytes in the dorsomedial hypothalamus accelerates sevoflurane anesthesia emergence in mice

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