GRIK2过表达对七氟烷暴露幼鼠空间学习和记忆能力的改善作用及其机制

doi:10.13481/j.1671-587X.20260105

吉林大学学报(医学版) ›› 2026, Vol. 52 ›› Issue (1): 35-43.doi: 10.13481/j.1671-587X.20260105

GRIK2过表达对七氟烷暴露幼鼠空间学习和记忆能力的改善作用及其机制

田雨禾^1,^2,^3,⁴,张婧彬^1,^3,⁴,李群涛^1,^3,⁴,马莹芳^2,^3,⁴,高娃^2,^3,⁴,马克涛^1,^2,^3,⁴,司军强^1,^2,^3,⁴(),殷姜文^1,^2,^3,⁴()

^1.石河子大学第一附属医院麻醉科，新疆石河子 832000
^2.石河子大学医学院生理学教研室，新疆石河子 832000
^3.中亚高发病防治中心重点实验室，新疆石河子 832000
^4.新疆地方病和民族病重点实验室，新疆石河子 832000

收稿日期:2025-03-25 接受日期:2025-05-14 出版日期:2026-01-28 发布日期:2026-02-24
通讯作者: 司军强,殷姜文 E-mail:sijunqiang@shzu.edu.cn;yjw6654328@163.com
作者简介:田雨禾（1997-），女，广东省深圳市人，在读硕士研究生，主要从事全麻药物发育毒性方面的研究。
基金资助:
国家自然科学基金项目(82160235);石河子大学第一附属医院博士基金项目(BS202101)

Improvement effect of GRIK2 overexpression on spatial learning and memory ability of sevoflurane-exposed young rats and its mechanism

Yuhe TIAN^1,^2,^3,⁴,Jingbing ZHANG^1,^3,⁴,Quntao LI^1,^3,⁴,Yingfang MA^2,^3,⁴,Wa GAO^2,^3,⁴,Ketao MA^1,^2,^3,⁴,Junqiang SI^1,^2,^3,⁴(),Jiangwen YIN^1,^2,^3,⁴()

^1.Department of Anesthesiology，First Affiliated Hospital，Shihezi University，Shihezi 832000，China
^2.Department of Physiology，School of Medicine Shihezi University，Shihezi 832000，China
^3.National Health Commission Key Laboratory of Prevention and Treatment of Central Asia High Incidence Diseases，Shihezi 832000，China
^4.Key Laboratory of Xinjiang Endemic and Ethnic Diseases，Shihezi 832000，China

Received:2025-03-25 Accepted:2025-05-14 Online:2026-01-28 Published:2026-02-24
Contact: Junqiang SI,Jiangwen YIN E-mail:sijunqiang@shzu.edu.cn;yjw6654328@163.com

摘要/Abstract

摘要：

目的探讨过表达红藻氨酸受体亚基（GluK）2对于七氟烷（Sevo）暴露幼鼠空间学习和记忆能力的改善作用，并阐明其可能的分子机制。方法将44只C57BL/6J新生仔鼠随机分为对照组、Sevo组、Sevo+OE-NC组（病毒空载组）和Sevo+OE-GRIK2组（GRIK2过表达组）。分子实验每组3只，行为学实验每组8只。Morris水迷宫实验检测各组幼鼠的逃避潜伏期、在目标象限停留时间和穿越隐藏平台次数。仔鼠于出生后第6天（P6）构建Sevo麻醉模型，免疫荧光法观察各组幼鼠海马组织中GluK2蛋白表达情况及病毒转染情况。Western blotting法检测各组幼鼠海马组织中钠钾氯转运体1（NKCC1）、钾氯共转运体2（KCC2）和GluK2蛋白表达情况。结果 Morris水迷宫实验训练第3、4和5天，与对照组比较，Sevo组幼鼠逃避潜伏期明显延长（P<0.05或P<0.01）。Morris水迷宫实验训练第4和5天，与Sevo组比较，Sevo+OE-GRIK2组幼鼠逃避潜伏期明显缩短（P<0.05或P<0.01）；与Sevo+OE-NC组比较，Sevo+OE-GRIK2组幼鼠逃避潜伏期明显缩短（P<0.05或P<0.01）。与对照组比较，Sevo组幼鼠在目标象限停留时间减少（P<0.05）；与Sevo组比较，Sevo+OE-GRIK2组幼鼠在目标象限停留时间增加（P<0.01）；与Sevo+OE-NC组比较，Sevo+OE-GRIK2组幼鼠在目标象限停留时间增加（P<0.01）。与对照组比较，Sevo组幼鼠穿越隐藏平台次数减少（P<0.001）；与Sevo组比较，Sevo+OE-GRIK2组幼鼠穿越隐藏平台次数增加（P<0.001）；与Sevo+OE-NC组比较，Sevo+OE-GRIK2组幼鼠穿越隐藏平台次数增加（P<0.001）。免疫荧光法，与对照组比较，Sevo组幼鼠海马组织中GluK2蛋白荧光强度降低（P<0.05）；与Sevo组比较，Sevo+OE-GRIK2组幼鼠海马组织中GluK2蛋白荧光强度升高（P<0.01）；与Sevo+OE-NC组比较，Sevo+OE-GRIK2组幼鼠海马组织中GluK2蛋白荧光强度升高（P<0.01）。Western blotting法，与对照组比较，Sevo组幼鼠海马组织中KCC2和GluK2蛋白表达水平明显降低（P<0.05或P<0.001），NKCC1/KCC2比值明显升高（P<0.05）；与Sevo组比较，Sevo+OE-GRIK2组幼鼠海马组织中KCC2和GluK2蛋白表达水平升高（P<0.001），NKCC1/KCC2比值降低（P<0.05）；与Sevo+OE-NC组比较，Sevo+OE-GRIK2组幼鼠海马组织中KCC2和GluK2蛋白表达水平升高（P<0.001），NKCC1/KCC2比值降低（P<0.05）。结论 GRIK2过表达使Sevo暴露的仔鼠海马组织中GluK2和KCC2蛋白表达上调，改善幼鼠空间学习和记忆能力，其机制可能与降低海马组织中NKCC1/KCC2比值有关。

关键词: 七氟烷, 全麻药物, 红藻氨酸受体亚基2, 钠钾氯转运体1, 钾氯共转运体2

Abstract:

Objective To discuss the improvement effect of overexpressing kainate receptor subunit （GluK） 2 on the spatial learning and memory abilities of juvenile mice exposed to sevoflurane （Sevo）， and to clarify its possible molecular mechanism. Methods A total of 44 C57BL/6J neonatal mice were randomly divided into control group， Sevo group， Sevo+OE-NC group （virus empty vector group）， and Sevo+OE-GRIK2 group （GRIK2 overexpression group）. There were 3 mice in each group for molecular experiments and 8 mice in each group for behavioral experiments. Morris water maze test was used to detect the escape latency， the residence time in the target quadrant， and the number of platform crossings of the juvenile mice in various groups. The Sevo anesthesia model was established in the mice on postnatal day 6 （P6）； immunofluorescence method was used to observe the expression of GluK2 protein and the virus transfection situation in hippocampus tissue of the juvenile mice in various groups； Western blotting method was used to detect the expression levels of sodium-potassium-chloride cotransporter 1 （NKCC1）， potassium-chloride cotransporter 2 （KCC2）， and GluK2 proteins in hippocampus tissue of the juvenile mice in various groups. Results The Morris water maze test results showed that on days 3， 4， and 5 of training， compared with control group， the escape latency of the young mice in Sevo group was significantly prolonged （P<0.05 or P<0.01）. The Morris water maze test results showed that on days 4 and 5 of training， compared with Sevo group， the escape latency of the young mice in Sevo+OE-GRIK2 group was significantly shortened （P<0.05 or P<0.01）； compared with Sevo+OE-NC group， the escape latency of the young mice in Sevo+OE-GRIK2 group was significantly shortened （P<0.05 or P<0.01）. The Morris water maze test results showed that compared with control group， the residence time in the target quadrant of the young mice in Sevo group was decreased （P<0.05）； compared with Sevo group， the residence time in the target quadrant of the young mice in Sevo+OE-GRIK2 group was increased （P<0.01）； compared with Sevo+OE-NC group， the residence time in the target quadrant of the young mice in Sevo+OE-GRIK2 group was increased （P<0.01）. The Morris water maze test results showed that compared with control group， the number of platform crossings of the young mice in Sevo group was decreased （P<0.001）； compared with Sevo group， the number of platform crossings of the young mice in Sevo+OE-GRIK2 group was increased （P<0.001）； compared with Sevo+OE-NC group， the number of platform crossings of the young mice in Sevo+OE-GRIK2 group was increased （P<0.001）. The immunofluorescence results showed that compared with control group， the expression level of GluK2 protein in hippocampus tissue of the young mice in Sevo group was decreased （P<0.05）； compared with Sevo group， the expression level of GluK2 protein in hippocampus tissue of the young mice in Sevo+OE-GRIK2 group was increased （P<0.01）； compared with Sevo+OE-NC group， the expression level of GluK2 protein in hippocampus tissue of the young mice in Sevo+OE-GRIK2 group was increased （P<0.01）. The Western blotting results showed that compared with control group， the expression levels of KCC2 and GluK2 proteins in hippocampus tissue of the young mice in Sevo group were decreased （P<0.05 or P<0.01）， and the NKCC1/KCC2 ratio was increased （P<0.05）； compared with Sevo group， the expression levels of KCC2 and GluK2 proteins in hippocampus tissue of the young mice in Sevo+OE-GRIK2 group were increased （P<0.001）， and the NKCC1/KCC2 ratio was decreased （P<0.05）； compared with Sevo+OE-NC group， the expression levels of KCC2 and GluK2 proteins in hippocampus tissue of the young mice in Sevo+OE-GRIK2 group were increased （P<0.001）， and the NKCC1/KCC2 ratio was decreased （P<0.05）. Conclusion Overexpression of GRIK2 upregulates the expression levels of GluK2 and KCC2 proteins in hippocampus tissue of Sevo-exposed juvenile mice and improves the spatial learning and memory abilities of juvenile mice， and its mechanism may be related to reducing the NKCC1/KCC2 ratio in hippocampus tissue.

Key words: Sevoflurane, General anesthetics, Kainate receptor subunit 2, Sodium-potassium-chloride cotransporter 1, Potassium-chloride cotransporter 2

中图分类号:

R965

田雨禾,张婧彬,李群涛,马莹芳,高娃,马克涛,司军强,殷姜文. GRIK2过表达对七氟烷暴露幼鼠空间学习和记忆能力的改善作用及其机制[J]. 吉林大学学报(医学版), 2026, 52(1): 35-43.

Yuhe TIAN,Jingbing ZHANG,Quntao LI,Yingfang MA,Wa GAO,Ketao MA,Junqiang SI,Jiangwen YIN. Improvement effect of GRIK2 overexpression on spatial learning and memory ability of sevoflurane-exposed young rats and its mechanism[J]. Journal of Jilin University(Medicine Edition), 2026, 52(1): 35-43.

图/表 7

图1

表1

图2

表2

图3

图4

图5

参考文献 26

[1]	ZHANG J H， DENG H X， HUANG X L， et al. Pre-school children single inhalation anesthetic exposure and neuro-psychological development： a prospective study and Mendelian randomization analysis［J］. Front Neurol， 2024， 15： 1389203.
[2]	POOR ZAMANY NEJAT KERMANY M， ROODNESHIN F， AHMADI DIZGAH N， et al. Early childhood exposure to short periods of sevoflurane is not associated with later， lasting cognitive deficits［J］. Paediatr Anaesth， 2016， 26（10）： 1018-1025.
[3]	WARNER D O， ZACCARIELLO M J， KATUSIC S K， et al. Neuropsychological and behavioral outcomes after exposure of young children to procedures requiring general anesthesia： the mayo anesthesia safety in kids （MASK） study［J］. Anesthesiology， 2018， 129（1）： 89-105.
[4]	王俊宽，许柯炎，秦瑞鸿，等. 七氟烷后处理对脑缺血再灌注成年大鼠远期神经功能的影响［J］. 郑州大学学报（医学版）， 2024， 59（2）： 178-181.
[5]	刘宝林，曹维福. SIRT1介导的铁死亡在老年小鼠术后认知功能障碍中的作用及其机制［J］. 解放军医学杂志，2025， 50（3）： 332-340.
[6]	KOK M， SINGH I， AIZENMAN E， et al. Inefficient maturation of disease-linked mutant forms of the KCC2 potassium-chloride cotransporter correlates with predicted pathogenicity［J］. J Biol Chem， 2025， 301（4）： 108399.
[7]	PRESSEY J C， MAHADEVAN V， KHADEMULLAH C S， et al. A kainate receptor subunit promotes the recycling of the neuron-specific K⁺-Cl^- co-transporter KCC2 in hippocampal neurons［J］. J Biol Chem， 2017， 292（15）： 6190-6201.
[8]	BRANCHEREAU P， CATTAERT D. Chloride homeostasis in developing motoneurons［J］. Adv Neurobiol， 2022， 28： 45-61.
[9]	BLAESSE P， AIRAKSINEN M S， RIVERA C， et al. Cation-chloride cotransporters and neuronal function［J］. Neuron， 2009， 61（6）： 820-838.
[10]	GALANOPOULOU A S. Developmental patterns in the regulation of chloride homeostasis and GABA（A） receptor signaling by seizures［J］. Epilepsia， 2007， 48（）： 14-18.
[11]	LIANG P， LI F， LIU J， et al. Sevoflurane activates hippocampal CA3 kainate receptors （Gluk2） to induce hyperactivity during induction and recovery in a mouse model［J］. Br J Anaesth， 2017， 119（5）： 1047-1054.
[12]	周斌，胡镐申，陈玲敏，等.腺相关病毒介导的小鼠发育期大脑中的特定细胞和特定脑区的基因编辑及潜在应用探索［J］. 临床和实验医学杂志， 2019， 18（3）： 225-229.
[13]	SUN M Y， XIE Z C， ZHANG J Q， et al. Mechanistic insight into sevoflurane-associated developmental neurotoxicity［J］. Cell Biol Toxicol， 2022， 38（6）： 927-943.
[14]	JEVTOVIC-TODOROVIC V， ABSALOM A R， BLOMGREN K， et al. Anaesthetic neurotoxicity and neuroplasticity： an expert group report and statement based on the BJA Salzburg Seminar［J］. Br J Anaesth， 2013， 111（2）： 143-151.
[15]	HOU Q， YUAN J H， LI S， et al. Autophagic degradation of DHCR7 activates AKT3 and promotes sevoflurane-induced hippocampal neuroinflammation in neonatal mice［J］. Free Radic Biol Med， 2024， 222： 304-316.
[16]	WANG S， LI Z J， LIU X， et al. Repeated postnatal sevoflurane exposure impairs social recognition in mice by disrupting GABAergic neuronal activity and development in hippocampus［J］. Br J Anaesth， 2024， 133（4）： 810-822.
[17]	ZHANG W H， CHEN Y X， QIN J W， et al. Prolonged sevoflurane exposure causes abnormal synapse development and dysregulates beta-neurexin and neuroligins in the hippocampus in neonatal rats［J］. J Affect Disord， 2022， 312： 22-29.
[18]	ZHOU B， CHEN L M， LIAO P， et al. Astroglial dysfunctions drive aberrant synaptogenesis and social behavioral deficits in mice with neonatal exposure to lengthy general anesthesia［J］. PLoS Biol， 2019， 17（8）： e3000086.
[19]	CABRERA O H， TESIC V， TAT Q L， et al. Sevoflurane-induced dysregulation of cation-chloride cotransporters NKCC1 and KCC2 in neonatal mouse brain［J］. Mol Neurobiol， 2020， 57（1）： 1-10.
[20]	ZHU Y W， ARMSTRONG J N， CONTRACTOR A. Kainate receptors regulate the functional properties of young adult-born dentate granule cells［J］. Cell Rep， 2021， 36（12）： 109751.
[21]	DUAN G F， TANG X H， JIA M， et al. Kainate receptors GluK1 and GluK2 differentially regulate synapse morphology［J］. Synapse， 2023， 77（1）： e22255.
[22]	TAKEISHI Y， YAGUCHI H， KUDO A， et al. The importance of early immunotherapy in anti-GluK2 antibody-positive autoimmune cerebellar ataxia： a case report［J］. J Neurol Sci， 2024， 467： 123306.
[23]	STOLZ J R， FOOTE K M， VEENSTRA-KNOL H E， et al. Clustered mutations in the GRIK2 kainate receptor subunit gene underlie diverse neurodevelopmental disorders［J］. Am J Hum Genet， 2021， 108（9）： 1692-1709.
[24]	WATANABE M， FUKUDA A. Development and regulation of chloride homeostasis in the central nervous system［J］. Front Cell Neurosci， 2015， 9： 371.
[25]	RIVERA C， VOIPIO J， PAYNE J A， et al. The K⁺/Cl^- co-transporter KCC2 renders GABA hyperpolarizing during neuronal maturation［J］. Nature， 1999， 397（6716）： 251-255.
[26]	MAHADEVAN V， PRESSEY J C， ACTON B A， et al. Kainate receptors coexist in a functional complex with KCC2 and regulate chloride homeostasis in hippocampal neurons［J］. Cell Rep， 2014， 7（6）： 1762-1770.

Group	Escape latency
Group	Day 1	Day 2	Day 3	Day 4	Day 5
Control	52.14±3.10	43.38±5.62	33.27±4.48	27.53±5.10	13.26±5.20
Sevo	51.01±4.55	44.23±4.10	40.73±2.93^**	36.12±5.88^**	21.69±5.05^*
Sevo+OE-NC	51.41±4.64	44.31±4.64	42.60±3.93	35.46±3.49	24.52±4.84
Sevo+OE-GRIK2	50.70±4.30	41.05±4.10	37.51±5.33	28.75±3.96^△△##	16.15±5.70^△#

Group	Residence time intarget quadrant (t/s)	Number of platform crosssing
Control	23.20±3.50	5.63±1.06
Sevo	18.16±4.42^*	3.15±1.13^**
Sevo+OE-NC	19.31±3.76	2.23±0.83
Sevo+OE-GRIK2	27.56±5.85^△#	5.50±1.31^△△##

GRIK2过表达对七氟烷暴露幼鼠空间学习和记忆能力的改善作用及其机制

Improvement effect of GRIK2 overexpression on spatial learning and memory ability of sevoflurane-exposed young rats and its mechanism

RichHTML

PDF (PC)

赞

可视化

摘要/Abstract

引用本文

使用本文

图/表 7

参考文献 26

相关文章 2

Metrics

本文评价

推荐阅读 0

[1]	陈芳, 马星钢, 孟保英, 赵雷. 七氟烷预处理在婴幼儿体外循环中对肺功能的保护作用[J]. 吉林大学学报(医学版), 2016, 42(04): 793-797.
[2]	申雪花, 韩伟, 王金, 麻海春. 七氟烷吸入麻醉对腹部手术患者血清肌钙蛋白I的影响[J]. J4, 2009, 35(3): 539-542.