Protective effect and mechanism of heat acclimation on hippocampus neuron injury in mice after exposure to electromagnetic field

WANG Zeze; YANG Xuesen; WANG Ying; TAN Yulong; LUO Zhen; LI Ping; HE Genlin; LIU Xiaoqian; SHEN Tingting; LIU Yishan; LUO Xue

doi:10.16016/j.2097-0927.202408087

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Protective effect and mechanism of heat acclimation on hippocampus neuron injury in mice after exposure to electromagnetic field

Neuroscience | 更新时间：2025-04-14

- Protective effect and mechanism of heat acclimation on hippocampus neuron injury in mice after exposure to electromagnetic field
- Journal of Army Medical University Vol. 47, Issue 7, Pages: 629-638(2025)
- 作者机构：
  
  1.陆军军医大学（第三军医大学）：军事预防医学系热带医学教研室，（培育），重庆
  2.陆军军医大学（第三军医大学）：极端环境医学教育部重点实验室，（培育），重庆
- 作者简介：
  
  LUO Xue， E-mail：luoxuecq@hotmail.com
- 基金信息：
- DOI：10.16016/j.2097-0927.202408087
  CLC： R339.52;R363.22;R594.801
- Received：20 August 2024，
  
  Revised：20 February 2025，
  
  Published：15 April 2025
- 稿件说明：
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王泽泽,杨学森,王英,等.热习服对电磁辐射所致小鼠海马神经损伤的保护作用及机制研究[J].陆军军医大学学报,2025,47(07):629-638.

WANG Zeze,YANG Xuesen,WANG Ying,et al.Protective effect and mechanism of heat acclimation on hippocampus neuron injury in mice after exposure to electromagnetic field[J].J Army Med Univ,2025,47(07):629-638.
王泽泽,杨学森,王英,等.热习服对电磁辐射所致小鼠海马神经损伤的保护作用及机制研究[J].陆军军医大学学报,2025,47(07):629-638. DOI：10.16016/j.2097-0927.202408087.

WANG Zeze,YANG Xuesen,WANG Ying,et al.Protective effect and mechanism of heat acclimation on hippocampus neuron injury in mice after exposure to electromagnetic field[J].J Army Med Univ,2025,47(07):629-638. DOI：10.16016/j.2097-0927.202408087.

摘要

目的

探究热习服对电磁辐射所致海马神经损伤的保护作用及机制。

方法

40只雄性BALB/c小鼠（体质量18~22 g，7周龄）按随机数字表法分为4组（

=10）：对照（Con）组、热习服（HA）组（34 ℃，30 d）、电磁辐射（EMF）组（2 450 MHz，20 min/d，4周）、热习服+电磁辐射（HA+EMF）组。采用糖水偏好实验检测各组小鼠蔗糖偏好程度，悬尾实验和强迫游泳实验观察各组不动时间，Morris水迷宫检测各组学习记忆能力。采用HE染色观察各组小鼠海马组织病理变化；分别使用小胶质细胞标志物Iba1、促炎表型标志物CD68及抗炎表型标志物CD206进行免疫荧光标记，检测各组海马内小胶质细胞数量及小胶质细胞表型变化；采用ELISA检测各组海马内TNF-α、IL-1β、TGF-β和IL-10水平；采用Western blot检测各组海马内HSP70的蛋白表达水平。

结果

与Con组相比，EMF暴露小鼠对蔗糖的偏好降低（

0.05），悬尾不动时间延长（

0.01），强迫游泳不动时间延长（

0.01），第7天的逃避潜伏期延长（

0.01），穿越平台次数减少（

0.05）；海马区细胞排列较紊乱、结构较松散、存在细胞形态不规则、肿胀，细胞核固缩、着色深，小胶质细胞水肿；海马区小胶

质细胞数量增加（

0.01），CD68相对荧光强度升高（

0.01）、CD206相对荧光强度差异无统计学意义（

=0.885），提示此时活化小胶质细胞主要呈促炎型M1表型；海马中TNF-α、IL-1β含量显著升高，IL-10和TGF-β含量显著下降（

0.01）。与EMF组相比，HA+EMF组小鼠对蔗糖的偏好增加（

0.01），悬尾不动时间减少（

0.05），强迫游泳不动时间减少（

0.01），第7天的逃避潜伏期缩短（

0.01）；小鼠海马区细胞损伤程度改善，细胞固缩减轻。与Con组相比，HA+EMF组海马区小胶质细胞数量增加（

0.01），抗炎型M2表型标志物CD206相对荧光强度升高（

0.01），此时CD68相对荧光强度相较EMF组显著下降（

0.01）。与EMF组相比，HA+EMF组小鼠海马中TNF-α、IL-1β水平下降（

0.01），TGF-β和IL-10水平升高（

0.01）。HA+EMF组小鼠海马内HSP70蛋白表达量相较于EMF组显著升高（

0.01）且显著高于HA组（

0.01）。

结论

热习服可能通过改变小胶质细胞表型调节炎症反应，改善电磁辐射诱导的神经损伤。

Abstract

Objective‍　‍To investigate the protective effect and mechanism of heat acclimation (HA) on electromagnetic field (EMF) induced hippocampus neuron injury in mice. Methods‍　‍Forty healthy BALB/c male mice (18~22 g

7 weeks old) were randomly divided into 4 groups (

=10): Control group (Con)

HA group (34 ℃

30 d)

EMF group (2 450 MHz

20 min/d

4 weeks) and HA+EMF group (HA preconditioning+EMF). Sucrose preference test was performed to evaluate sucrose preference levels of mice in each group. Tail suspension test and forced swimming test were utilized to observe the immobility time. Morris water maze test was conducted to determine the learning and memory capabilities. Pathological changes in the hippocampus were observed with HE staining. Immunohistochemical assay for Iba1 (marker of microglia)

CD68 (marker of pro-inflammatory phenotype) and CD206 (marker of anti-inflammatory phenotype) were used to detect the number and activation phenotype of microglia in the hippocampus. ELISA was applied to measure the levels of TNF-α

IL-1β

TGF-β and IL-10 in the hippocampus of each group. Western blott

ing was performed to determine the protein levels of HSP70 in the hippocampus. Results‍　‍As compared with the Con group

the EMF group showed a decreased preference for sucrose (

0.05)

prolonged immobile time in the tail suspension test (

0.01) as well as in the forced swimming test (

0.01)

extented escape latency on the 7th day (

0.01)

and a decreased time of crossing the platform (

0.05). EMF exposure resulted in that the hippocampal neurons were in disordered arrangement

loose structure and irregular morphology

with swollen cytoplasm and condensed nuclei

swollen and more microglial cells in the hippocampus (

0.01)

and enhanced relative fluorescence intensity of CD68 (

0.01)

but not in CD206 fluorescence intensity (

=0.885). All these findings suggested that activated microglia predominantly exhibited a pro-inflammatory M1 phenotype during this phase. In the hippocampus

the levels of TNF-α and IL-1β were significantly increased

while the levels of IL-10 and TGF-β were significantly decreased (

0.01).HA treatment reversed the conditions induced by EMF exposure

including better preference for sucrose (

0.01)

shorten immobile time in tail suspension test (

0.05) and forced swimming test (

0.01)

less escape latency on the 7th day (

0.01)

and improved hippocampal cell injuries. Compared with the Con group

there were more microglial cells in the hippocampus in the HA+EMF group

with increased relative fluorescence intensity of M2 phenotype marker CD206 (

0.01) and decreased CD68 fluorescence intensity (

0.01). HA treatment also significantly decreased the expression of TNF-α and IL-1β levels (

0.01)

increased the expression of IL-10 and TGF-β (

0.01)

and

elevated the protein level of HSP70 (

0.01) when compared with the EMF group. Conclusion‍　‍HA may ameliorate EMF-induced hippocampus neurons injury in mice by altering the phenotype of activated microglia and inhibiting inflammatory responses.

关键词

Keywords

references

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