Article(id=1322192315049951925, tenantId=1146029695717560320, journalId=1319373162502258839, issueId=1322192284670603597, articleNumber=null, orderNo=null, doi=10.16016/j.2097-0927.202504048, pmid=null, cstr=null, oa=null, hot=null, price=null, onlineType=0, articleFormat=0, articleType=null, articleTypeStr=null, receivedDate=1744387200000, receivedDateStr=2025-04-12, revisedDate=1750521600000, revisedDateStr=2025-06-22, acceptedDate=null, acceptedDateStr=null, onlineDate=1793164813053, onlineDateStr=2026-10-28, pubDate=1755187200000, pubDateStr=2025-08-15, doiRegisterDate=null, doiRegisterDateStr=null, onlineIssueDate=1793164813053, onlineIssueDateStr=2026-10-28, onlineJustAcceptDate=null, onlineJustAcceptDateStr=null, onlineFirstDate=null, onlineFirstDateStr=null, sourceXml=null, magXml=null, createTime=1793164813053, creator=13701087609, updateTime=1793164813053, updator=13701087609, issue=Issue{id=1322192284670603597, tenantId=1146029695717560320, journalId=1319373162502258839, year='2025', volume='47', issue='15', pageStart='1685', pageEnd='1824', issueExtLink='null', onlineDate='null', pubDate='1755187200000', pubDateStr='2025-08-15', beforeIssueId=null, nextIssueId=null, price=null, status=1, issueComplete=1, articleOrder=1, issueType=-1, specialIssue=0, createTime=1793164805810, creator='13701087609', updateTime=1793164826583, updator='13701087609', preIssue=null, nextIssue=null, articleTotal=null, ext={EN=IssueExt(id=1322192371861799855, tenantId=1146029695717560320, journalId=1319373162502258839, issueId=1322192284670603597, language=EN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=), CN=IssueExt(id=1322192371861799856, tenantId=1146029695717560320, journalId=1319373162502258839, issueId=1322192284670603597, language=CN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=)}, issueFiles=null, downloadFileDto=null}, startPage=1815, endPage=1824, ext={EN=ArticleExt(id=1322192315750400694, articleId=1322192315049951925, tenantId=1146029695717560320, journalId=1319373162502258839, language=EN, title=Effects of long-term 2.65 GHz radiofrequency radiation on inflammatory response and intestinal microbiota in mice, columnId=1320435151990014650, journalTitle=Journal of Army Medical University, columnName=Military Medicine, runingTitle=null, highlight=null, articleAbstract=
Objective‍

To investigate the effects of long-term radiofrequency (RF) radiation at 2.65 GHz on behavior, inflammatory response, and intestinal microecology in mice in order to provide a basis for the safety assessment of long-term RF exposure.

Methods‍

One hundred and eight male C57BL/6N mice (17~21 g, 6~8 weeks old) were randomly assigned to a control group (Con) and a RF exposure group. The mice of the RF exposure group were subjected to whole-body uniform exposure to 2.65 GHz RF radiation in an electromagnetic reverberation chamber for 3 h/day for 28 consecutive days. RF field distribution and changes in core body temperature were monitored using an electromagnetic radiation analyzer and a fiber-optic temperature probe, respectively. Cognitive function was assessed using the Y-maze and novel object recognition (NOR) test. Anxiety-like behaviors were evaluated through open field test (OFT) and elevated plus maze (EPM), while depressive-like behaviors were examined with sucrose preference test (SPT) and tail suspension test (TST). HE staining was used to observe the histopathological changes in mouse tissues. Radioimmunoassay (RIA) was employed to detect the expression of pro-inflammatory cytokines, TNF-α and IL-1β, as well as anti-inflammatory cytokines, IL-4 and IL-10 in the serum, brain, jejunum, and spleen samples. Additionally, metagenomic sequencing was performed to assess alterations in the gut microbiota composition.

Results‍

Long-term RF radiation led to a maximal increase of 0.59 ℃ in the core body temperature, but had no significant effects on cognitive function, anxiety-like behaviors, or depressive-like behaviors, or apparent damage of the hippocampal or jejunal tissues in the exposed mice. However, RF exposure significantly up-regulated the expression of the pro-inflammatory cytokine TNF-α in serum (P<0.05), and did not significantly alter the concentrations of other cytokines (IL-1β,IL-4,IL-10), caused significant decrease in α-diversity of the intestinal microbiota (P<0.01), with reduced relative abundances of Ligilactobacillus murinus and Acetatifactor muris (P<0.05), while elevated abundances of Lachnospiraceae bacterium (P<0.01).

Conclusion‍

Long-term exposure to 2.65 GHz RF radiation induces systemic inflammatory responses and disrupts gut microbiota homeostasis in mice.

, authors=Keqin LI1, 2, Yanhui HAO2, Ying LIU2, Jun WANG2, Hongyan ZUO2, Hong YANG1, Yang LI2, Hua DENG1, authorsList=Keqin LI, Yanhui HAO, Ying LIU, Jun WANG, Hongyan ZUO, Hong YANG, Yang LI, Hua DENG, authorCompany=null, correspAuthors=Hua DENG, authorNote=null, correspAuthorsNote=null, copyrightStatement=© 2025 Journal of Army Medical University, copyrightOwner=Editorial Office of Journal of Army Medical University, extLink=null, articleAbsUrl=null, sourceXml=null, magXml=null, pdfUrl=null, pdf=null, pdfFileSize=null, pdfExtLink=null, richHtmlUrl=null, mobilePdfUrl=null, reviewReport=null, pdfFirstPage=null, abstractGraph=null, abstractGraphContent=null, abstractVideo=null, citation=null, cebUrl=null, magXmlContent=null, mapNumber=null, fund=null), CN=ArticleExt(id=1322192319097455303, articleId=1322192315049951925, tenantId=1146029695717560320, journalId=1319373162502258839, language=CN, title=2.65 GHz射频长期辐射对小鼠机体炎症反应和肠道微生态的影响, columnId=1322185581656920707, journalTitle=陆军军医大学学报, columnName=军事医学, runingTitle=null, highlight=null, articleAbstract=
目的

探讨2.65 GHz长期射频辐射(radiofrequency radiation,RFR)对小鼠行为学、机体炎症反应和肠道微生态的影响,为射频辐射长期暴露的安全性评估提供依据。

方法

108只C57BL/6N雄性小鼠(体质量17~21 g,6~8周龄)按照随机数字表法分为对照(Con)组和RFR组;通过电磁混响室模拟2.65 GHz射频辐射场对小鼠进行全身均匀暴露,时间为3 h/d,持续28 d;采用电磁辐射分析仪和光纤测温仪检测射频辐射场强分布和小鼠核心体温的变化;Y-迷宫和新物体识别实验检测射频辐射对小鼠认知功能的影响;旷场和高架十字迷宫实验检测射频辐射对小鼠焦虑情绪的影响;糖水偏好和悬尾实验检测射频辐射对小鼠抑郁情绪的影响;HE染色观察小鼠组织结构;放射免疫法检测小鼠血清、脑、空肠和脾脏中促炎因子肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)、白介素-1β(interleukin-1β,IL-1β)和抑炎因子白介素-4(interleukin-4,IL-4)、白介素-10(interleukin-10,IL-10)的表达;采用宏基因组测序技术检测小鼠肠道微生态的变化。

结果

本实验条件下长期射频辐射引起小鼠核心温度最大升幅达0.59 ℃;对小鼠认知、焦虑和抑郁情绪未见明显影响;对小鼠海马和空肠组织结构未见明显损伤;引起小鼠血清中促炎因子TNF-α表达显著上调(P<0.05),并未引起其他细胞因子(IL-1β、IL-4、IL-10)的含量改变;引起小鼠肠道菌群α多样性显著下降(P<0.01),在种水平上鼠乳酸杆菌(Ligilactobacillus murinus)和乙酰因子属菌(Acetatifactor muris,Acetatifactor aquisgranensis)相对丰度显著减少(P<0.05),毛螺科菌(Lachnospiraceae bacterium)相对丰度显著增多(P<0.01)。

结论

2.65 GHz长期射频辐射引起小鼠机体发生炎症反应以及肠道微生态紊乱。

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邓桦,E-mail:
DENG Hua,E-mail:
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Gut,2021,70(4):666-676., articleTitle=Angiogenin maintains gut microbe homeostasis by balancing α-Proteobacteria and Lachnospiraceae, refAbstract=null)], funds=null, companyList=[AuthorCompany(id=1322192319307170504, tenantId=1146029695717560320, journalId=1319373162502258839, articleId=1322192315049951925, xref=1, ext=[AuthorCompanyExt(id=1322192319315559113, tenantId=1146029695717560320, journalId=1319373162502258839, articleId=1322192315049951925, companyId=1322192319307170504, language=EN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=1School of Animal Science and Technology,Foshan University,Foshan,Guangdong), AuthorCompanyExt(id=1322192319323947722, tenantId=1146029695717560320, journalId=1319373162502258839, articleId=1322192315049951925, companyId=1322192319307170504, language=CN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=1佛山大学动物科技学院,广东 佛山)]), AuthorCompany(id=1322192319399445195, tenantId=1146029695717560320, journalId=1319373162502258839, articleId=1322192315049951925, xref=2, ext=[AuthorCompanyExt(id=1322192319407833804, tenantId=1146029695717560320, journalId=1319373162502258839, articleId=1322192315049951925, companyId=1322192319399445195, language=EN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=2Academy of Military Medical Sciences,Academy of Military Science,Beijing,China), AuthorCompanyExt(id=1322192319420416717, tenantId=1146029695717560320, journalId=1319373162502258839, articleId=1322192315049951925, companyId=1322192319399445195, language=CN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=2军事科学院军事医学研究院,北京)])], figs=[ArticleFig(id=1322192324029956865, tenantId=1146029695717560320, journalId=1319373162502258839, articleId=1322192315049951925, language=EN, label=null, caption=null, figureFileSmall=T3NSCfxL/Ht1iTAyGRDLJA==, figureFileBig=nUYu1T4buZ+A1Jaf4RhWbQ==, tableContent=null), ArticleFig(id=1322192324134814466, tenantId=1146029695717560320, journalId=1319373162502258839, articleId=1322192315049951925, language=CN, label=图1, caption=射频辐射有效工作区电场强度检测和小鼠体温变化(n=3,x¯±se)

A:ERC有效工作区不同点位电场强度测量结果;B:RFR前、中、后小鼠体温变化

, figureFileSmall=T3NSCfxL/Ht1iTAyGRDLJA==, figureFileBig=nUYu1T4buZ+A1Jaf4RhWbQ==, tableContent=null), ArticleFig(id=1322192324298392324, tenantId=1146029695717560320, journalId=1319373162502258839, articleId=1322192315049951925, language=EN, label=null, caption=null, figureFileSmall=vYuk4lcFGF1o3VyIYlqTDA==, figureFileBig=kylq4DHC/tZ1kwjTmH4Vxw==, tableContent=null), ArticleFig(id=1322192324369695493, tenantId=1146029695717560320, journalId=1319373162502258839, articleId=1322192315049951925, language=CN, label=图2, caption=长期射频辐射对小鼠空间和物体识别记忆的影响(n=10,x¯±se)

A~C:Y-迷宫自由交替实验的代表性运动轨迹热图、小鼠平均速度以及自由交替比;D~F:新物体识别实验的代表性运动轨迹图、新物体时间占比以及新物体辨别指数 红框:示新物体

, figureFileSmall=vYuk4lcFGF1o3VyIYlqTDA==, figureFileBig=kylq4DHC/tZ1kwjTmH4Vxw==, tableContent=null), ArticleFig(id=1322192324587799302, tenantId=1146029695717560320, journalId=1319373162502258839, articleId=1322192315049951925, language=EN, label=null, caption=null, figureFileSmall=CjajWCWO3xpY3v8fajYREg==, figureFileBig=oxpSakErbrHfUenxBYwzTQ==, tableContent=null), ArticleFig(id=1322192324646519559, tenantId=1146029695717560320, journalId=1319373162502258839, articleId=1322192315049951925, language=CN, label=图3, caption=长期射频辐射对小鼠焦虑行为的影响(n=10,x¯±se)

A~C:旷场实验的代表性运动轨迹图、中心区域时间以及中心区域次数;D~F:高架十字迷宫实验的代表性运动轨迹热图、开放臂时间占比以及开放臂次数占比

, figureFileSmall=CjajWCWO3xpY3v8fajYREg==, figureFileBig=oxpSakErbrHfUenxBYwzTQ==, tableContent=null), ArticleFig(id=1322192324969480969, tenantId=1146029695717560320, journalId=1319373162502258839, articleId=1322192315049951925, language=EN, label=null, caption=null, figureFileSmall=FKJEj3f8Hpx5lOD9XSvsiQ==, figureFileBig=VTrKrHezNKWe1h1DdKIAHA==, tableContent=null), ArticleFig(id=1322192325040784138, tenantId=1146029695717560320, journalId=1319373162502258839, articleId=1322192315049951925, language=CN, label=图4, caption=长期射频辐射对小鼠抑郁行为的影响(n=10,x¯±se)

A、B:糖水偏好实验的糖水偏好时间占比、糖水偏好指数;C、D:悬尾实验的不动时间占比、挣扎时间占比

, figureFileSmall=FKJEj3f8Hpx5lOD9XSvsiQ==, figureFileBig=VTrKrHezNKWe1h1DdKIAHA==, tableContent=null), ArticleFig(id=1322192325099504395, tenantId=1146029695717560320, journalId=1319373162502258839, articleId=1322192315049951925, language=EN, label=null, caption=null, figureFileSmall=R/M4EHZSRUviz7weS1mG5Q==, figureFileBig=fZ7/5ltX1ehw+BC2GATrpg==, tableContent=null), ArticleFig(id=1322192325162418956, tenantId=1146029695717560320, journalId=1319373162502258839, articleId=1322192315049951925, language=CN, label=图5, caption=长期射频辐射对小鼠海马(A)和空肠组织(B)结构的影响, figureFileSmall=R/M4EHZSRUviz7weS1mG5Q==, figureFileBig=fZ7/5ltX1ehw+BC2GATrpg==, tableContent=null), ArticleFig(id=1322192326919832333, tenantId=1146029695717560320, journalId=1319373162502258839, articleId=1322192315049951925, language=EN, label=null, caption=null, figureFileSmall=2cgEmPBuKIPhVST4kVVPKw==, figureFileBig=UWVdh7XFxewmyDhsYzumKA==, tableContent=null), ArticleFig(id=1322192327054050062, tenantId=1146029695717560320, journalId=1319373162502258839, articleId=1322192315049951925, language=CN, label=图6, caption=长期射频辐射对小鼠促炎因子(A、B)和抑炎因子(C、D)表达的影响(n=6,x¯±se)

a:P<0.05,与Con组比较

, figureFileSmall=2cgEmPBuKIPhVST4kVVPKw==, figureFileBig=UWVdh7XFxewmyDhsYzumKA==, tableContent=null), ArticleFig(id=1322192327125353231, tenantId=1146029695717560320, journalId=1319373162502258839, articleId=1322192315049951925, language=EN, label=null, caption=null, figureFileSmall=m1XGsb2IPTXk4dbH2zp3sA==, figureFileBig=6NC/MygirtaHGBufobwnng==, tableContent=null), ArticleFig(id=1322192327184073488, tenantId=1146029695717560320, journalId=1319373162502258839, articleId=1322192315049951925, language=CN, label=图7, caption=长期射频辐射对小鼠肠道菌群α多样性的影响(n=12,x¯±se)

a:P<0.001,b:P<0.01,与Con组比较

, figureFileSmall=m1XGsb2IPTXk4dbH2zp3sA==, figureFileBig=6NC/MygirtaHGBufobwnng==, tableContent=null), ArticleFig(id=1322192327238599441, tenantId=1146029695717560320, journalId=1319373162502258839, articleId=1322192315049951925, language=EN, label=null, caption=null, figureFileSmall=JeXfhbCjVhJTuGDqy5HTxw==, figureFileBig=DXQrUUBCVZwXc23kn7gG8g==, tableContent=null), ArticleFig(id=1322192327309902610, tenantId=1146029695717560320, journalId=1319373162502258839, articleId=1322192315049951925, language=CN, label=图8, caption=长期射频辐射对小鼠肠道微生物种水平前20相对丰度的影响

a:P<0.05,b:P<0.01,c:P<0.001,与Con组比较

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2.65 GHz射频长期辐射对小鼠机体炎症反应和肠道微生态的影响
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李刻勤 1, 2 , 郝延辉 2 , 刘莹 2 , 王俊 2 , 左红艳 2 , 杨鸿 1 , 李杨 2 , 邓桦 1
陆军军医大学学报 | 军事医学 2025,47(15): 1815-1824
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陆军军医大学学报 |军事医学 2025 , 47 (15) : 1815 -1824
2.65 GHz射频长期辐射对小鼠机体炎症反应和肠道微生态的影响
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李刻勤1, 2, 郝延辉2, 刘莹2, 王俊2, 左红艳2, 杨鸿1, 李杨2, 邓桦1
作者信息
  • 1佛山大学动物科技学院,广东 佛山
  • 2军事科学院军事医学研究院,北京
通讯作者:
邓桦,E-mail:
DENG Hua,E-mail:
Effects of long-term 2.65 GHz radiofrequency radiation on inflammatory response and intestinal microbiota in mice
Keqin LI1, 2, Yanhui HAO2, Ying LIU2, Jun WANG2, Hongyan ZUO2, Hong YANG1, Yang LI2, Hua DENG1
Affiliations
  • 1School of Animal Science and Technology,Foshan University,Foshan,Guangdong
  • 2Academy of Military Medical Sciences,Academy of Military Science,Beijing,China
出版时间: 2025-08-15 doi: 10.16016/j.2097-0927.202504048
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目的

探讨2.65 GHz长期射频辐射(radiofrequency radiation,RFR)对小鼠行为学、机体炎症反应和肠道微生态的影响,为射频辐射长期暴露的安全性评估提供依据。

方法

108只C57BL/6N雄性小鼠(体质量17~21 g,6~8周龄)按照随机数字表法分为对照(Con)组和RFR组;通过电磁混响室模拟2.65 GHz射频辐射场对小鼠进行全身均匀暴露,时间为3 h/d,持续28 d;采用电磁辐射分析仪和光纤测温仪检测射频辐射场强分布和小鼠核心体温的变化;Y-迷宫和新物体识别实验检测射频辐射对小鼠认知功能的影响;旷场和高架十字迷宫实验检测射频辐射对小鼠焦虑情绪的影响;糖水偏好和悬尾实验检测射频辐射对小鼠抑郁情绪的影响;HE染色观察小鼠组织结构;放射免疫法检测小鼠血清、脑、空肠和脾脏中促炎因子肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)、白介素-1β(interleukin-1β,IL-1β)和抑炎因子白介素-4(interleukin-4,IL-4)、白介素-10(interleukin-10,IL-10)的表达;采用宏基因组测序技术检测小鼠肠道微生态的变化。

结果

本实验条件下长期射频辐射引起小鼠核心温度最大升幅达0.59 ℃;对小鼠认知、焦虑和抑郁情绪未见明显影响;对小鼠海马和空肠组织结构未见明显损伤;引起小鼠血清中促炎因子TNF-α表达显著上调(P<0.05),并未引起其他细胞因子(IL-1β、IL-4、IL-10)的含量改变;引起小鼠肠道菌群α多样性显著下降(P<0.01),在种水平上鼠乳酸杆菌(Ligilactobacillus murinus)和乙酰因子属菌(Acetatifactor muris,Acetatifactor aquisgranensis)相对丰度显著减少(P<0.05),毛螺科菌(Lachnospiraceae bacterium)相对丰度显著增多(P<0.01)。

结论

2.65 GHz长期射频辐射引起小鼠机体发生炎症反应以及肠道微生态紊乱。

射频辐射  /  认知  /  情绪  /  炎症反应  /  肠道菌群
Objective‍

To investigate the effects of long-term radiofrequency (RF) radiation at 2.65 GHz on behavior, inflammatory response, and intestinal microecology in mice in order to provide a basis for the safety assessment of long-term RF exposure.

Methods‍

One hundred and eight male C57BL/6N mice (17~21 g, 6~8 weeks old) were randomly assigned to a control group (Con) and a RF exposure group. The mice of the RF exposure group were subjected to whole-body uniform exposure to 2.65 GHz RF radiation in an electromagnetic reverberation chamber for 3 h/day for 28 consecutive days. RF field distribution and changes in core body temperature were monitored using an electromagnetic radiation analyzer and a fiber-optic temperature probe, respectively. Cognitive function was assessed using the Y-maze and novel object recognition (NOR) test. Anxiety-like behaviors were evaluated through open field test (OFT) and elevated plus maze (EPM), while depressive-like behaviors were examined with sucrose preference test (SPT) and tail suspension test (TST). HE staining was used to observe the histopathological changes in mouse tissues. Radioimmunoassay (RIA) was employed to detect the expression of pro-inflammatory cytokines, TNF-α and IL-1β, as well as anti-inflammatory cytokines, IL-4 and IL-10 in the serum, brain, jejunum, and spleen samples. Additionally, metagenomic sequencing was performed to assess alterations in the gut microbiota composition.

Results‍

Long-term RF radiation led to a maximal increase of 0.59 ℃ in the core body temperature, but had no significant effects on cognitive function, anxiety-like behaviors, or depressive-like behaviors, or apparent damage of the hippocampal or jejunal tissues in the exposed mice. However, RF exposure significantly up-regulated the expression of the pro-inflammatory cytokine TNF-α in serum (P<0.05), and did not significantly alter the concentrations of other cytokines (IL-1β,IL-4,IL-10), caused significant decrease in α-diversity of the intestinal microbiota (P<0.01), with reduced relative abundances of Ligilactobacillus murinus and Acetatifactor muris (P<0.05), while elevated abundances of Lachnospiraceae bacterium (P<0.01).

Conclusion‍

Long-term exposure to 2.65 GHz RF radiation induces systemic inflammatory responses and disrupts gut microbiota homeostasis in mice.

radiofrequency radiation  /  cognition  /  emotion  /  inflammatory response  /  gut microbiota
李刻勤, 郝延辉, 刘莹, 王俊, 左红艳, 杨鸿, 李杨, 邓桦. 2.65 GHz射频长期辐射对小鼠机体炎症反应和肠道微生态的影响. 陆军军医大学学报, 2025 , 47 (15) : 1815 -1824 . DOI: 10.16016/j.2097-0927.202504048
Keqin LI, Yanhui HAO, Ying LIU, Jun WANG, Hongyan ZUO, Hong YANG, Yang LI, Hua DENG. Effects of long-term 2.65 GHz radiofrequency radiation on inflammatory response and intestinal microbiota in mice[J]. Journal of Army Medical University, 2025 , 47 (15) : 1815 -1824 . DOI: 10.16016/j.2097-0927.202504048
射频指频率为100 KHz~300 GHz的电磁波,广泛应用于通信、工业、军事和医疗等领域。近年来,随着互联网通信技术的高速发展,大众对通信网络的使用越来越普遍,接触射频辐射(radiofrequency radiation,RFR)的机会和程度不断加深。尤其是5G通信技术的应用和普及,表明射频技术正朝着更高频率的方向发展。根据中国互联网络信息中心发布的《第53次中国互联网络发展状况统计报告》显示,2023年互联网络人口占比达77.5%,我国网民人均一周内平均每天使用网络工具时长达3.5 h[1]。电磁辐射对人体健康的影响备受关注,既往电磁辐射生物效应研究多关注2G~4G等频段,关于5G频段RFR尤其是长期暴露对人体健康的潜在影响有待进一步评估[2]。
机体炎症反应在多种疾病的发生发展中发挥重要的作用,肠道微生物对炎症的调控作用逐渐受到重视。在RFR生物效应研究中,脑因为其重要性和敏感性而备受关注,低于机体发热阈值的射频暴露是否会对脑的功能产生不利影响尚存争议。有研究发现,RFR暴露导致小鼠脑组织中炎症因子表达上调和神经递质释放紊乱,从而引起小鼠认知异常和焦虑情绪[3]。此外,低强度辐射暴露可诱导大脑氧化应激,进而促进神经炎症反应等[4]。近年来,脑-肠轴对于神经系统功能发挥重要的调节作用,其中,肠道菌群对于物质代谢和炎症反应的调控作用逐渐受到关注[5-6]。有研究发现,4.9 GHz射频暴露可引起小鼠肠道微生物显著差异和代谢谱发生变化,并诱导小鼠抑郁样行为[7]。因此,基于人群接触射频电磁场的实际场景,在更长时间尺度上的RFR对脑认知和情绪等功能的影响进行评估,RFR是否引起机体发生炎症反应以及肠道微生态紊乱有待探讨[8]。
综上,本研究通过电磁混响室(electromagnetic reverberation chamber,ERC)模拟2.65 GHz RFR环境,对C57BL/6N小鼠进行全身均匀辐射。通过行为学、放射免疫和宏基因组测序等技术方法,研究长期RFR对小鼠认知和情绪、炎症调控相关因子表达以及肠道微生态的影响,基于肠道菌群对机体炎症反应的调控作用为长期RFR的安全性评估和防护提供理论依据。
实验采用SPF级C57BL/6N雄性小鼠(体质量17~21 g,6~8周龄),共108只,购自北京维通利华实验动物科技有限公司[许可证号:SCXK(京)2021-0006]。实验动物饲养于军事医学研究院实验动物中心,饲养条件:温度(22±2)℃,相对湿度50%~60%,光照/黑暗12/12 h,自由摄食和饮水。将小鼠按随机数字表法分为(n=54):Con组和RFR组。动物实验通过军事医学研究院实验动物管理与使用委员会批准(IACUC-DWZX-2023-P686)。
ERC暴露系统购自中国信息通信研究院环境与安全部;电磁辐射分析仪(NBM-550)购自德国Narda公司;光纤传感器、光纤测温仪(SPC-HR-2)购自加拿大FISO公司;旷场箱、新物体识别、Y-迷宫、悬尾、糖水偏好实验装置购自上海欣软信息科技有限公司;高架十字迷宫、增强型小动物气体麻醉仪、脑立体定位仪购自深圳瑞沃德生命科技有限公司;全景数字扫描系统(Pannoramic MIDI)购自匈牙利3DHISTECH公司;Any-maze行为学分析软件(版本7.16)购自美国Stoelting公司。
小鼠肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)、白介素-1β(interleukin-1β,IL-1β)、白介素-4(interleukin-4,IL-4)和白介素-10(interleukin-10,IL-10)放射免疫检测试剂盒购自北京福瑞润泽生物技术有限公司;福尔马林固定液、苏木素-伊红购自中国国药集团化学试剂有限公司;异氟烷购自深圳瑞沃德生命科技有限公司。
本研究采用2.65 GHz射频连续波作为射频辐射源,设置射频频率和电平值等参数。将RFR组小鼠放置于亚克力放射盒内进行固定,对放射盒进行打孔以保证散热,每天照射3 h,持续照射28 d。对Con组小鼠进行同等条件下的假照射。
ERC是由信号源产生2.65 GHz连续正弦波,并通过功率放大器进行放大,由射频发射天线将信号传输至双层铜网屏蔽体组成立方形混响室,混响室内布置电磁搅拌器[9]。场强检测:将NBM-550检测仪的探头放置于ERC有效工作区,并采用数据连接线将分析仪与电脑连接,通过NBM-TS软件实时记录和采集连续6 min电场强度的变化并计算平均值。通过Sim4Life软件进行建模和仿真计算,对小鼠RFR比吸收率(specific absorption rate,SAR)进行计算。
采用异氟烷对小鼠进行吸入麻醉,通过手术将光纤传感器探头埋置于小鼠腹腔内并缝合,待小鼠恢复至清醒状态并稳定30 min,接入光纤测温仪记录10 min作为照射前基底值,开始暴露3 h并持续观察记录小鼠温度,照射结束后关闭ERC照射设备并采集10 min作为小鼠照射后测温值。在数据处理时,计算每10分钟体温的均值,绘制折线图。
行为学实验开始前将小鼠放在行为室环境中适应,每天30 min,连续3 d;每只小鼠测试结束后,采用75%酒精对行为学箱进行擦拭并晾干,以避免残余排泄物和气味的影响。在测试过程中用布帘进行遮挡,保持环境安静。所有的行为学实验遵循以上习惯化程序。除糖水偏好实验在长期RFR暴露第28天检测外,其余行为学实验均在暴露第14天和第28天进行检测。
Y-迷宫自由交替实验:Y型迷宫装置由3个相同且互相呈120°的臂组成,分别作为A、B、C臂。每条臂长30 cm,宽8 cm,高15 cm。将小鼠放入Y-迷宫A臂,允许自由移动8 min。小鼠的4个爪子均进入臂内视为一次探索,小鼠连续进入3个不同臂,如ABC、BCA等,视作一次完整的交替行为。采用Any-maze软件记录及分析小鼠的交替行为和探索速度,计算自由交替比=连续进入3个不同臂的次数/(进臂总次数-2)×100%。
新物体识别实验:熟悉期,将小鼠放置在一个开放场地(40 cm×40 cm)中,自由探索5 min;适应期,在场地中对角放置2个相同颜色和材质的物体(视为A、B),并允许小鼠自由探索5 min。测试期,将新物体C(材质相同,但形状和颜色不同)替换掉物体B,并允许小鼠自由探索5 min。采用Any-maze软件记录及分析小鼠探索新物体的时间比和次数比。计算辨别指数=新物体/(新物体+熟悉物体)×100%。
旷场实验:旷场箱尺寸为40 cm×40 cm,将中央20 cm×20 cm区域设定为中心区域,其余为周围区域。测试时,将小鼠放置在旷场的中心区域,允许小鼠自由移动5 min。采用Any-maze软件记录及分析小鼠进入中心区的时间和次数。
高架十字迷宫实验:迷宫高出地面55 cm,分为呈十字交叉的开放臂和闭合臂(长65 cm,宽5 cm),闭合臂外围有高8 cm的挡板,而开放臂则无挡板。测试时,将小鼠放置在迷宫的中央平台上,头部面向开放臂,让小鼠自由探索迷宫5 min,并采用Any-maze软件记录及分析小鼠进入开放臂的时间占比和次数占比。
糖水偏好实验:熟悉阶段,采用2瓶饮用水进行6 h熟悉环节。适应阶段,采用2瓶体积相同的1%蔗糖溶液进行适应12 h,第二天将其中1瓶蔗糖溶液换成水溶液,持续24 h,为避免位置偏好,在第12小时将2瓶溶液互换位置,之后禁食禁水12 h。测试阶段,提供1瓶不含糖的饮用水和1瓶1%的蔗糖溶液,允许小鼠自由饮水,并记录3 h小鼠舔舐糖水的时间和次数。计算糖水偏好指数=舔舐糖水次数/(舔舐糖水次数+舔舐纯水次数)×100%。
悬尾实验:使用支撑杆和医用胶带将小鼠悬挂在离地面30 cm处,在距离小鼠尾巴尖端约2 cm处使用胶带将小鼠固定在吊杆上。录像设备持续记录小鼠6 min的运动情况,其中前2 min为适应阶段,记录后4 min的不动时间,并分析小鼠的不动时间占比和挣扎时间占比,在此期间只有前肢动,后肢不动,被认为是小鼠的摇摆行为(绝望状态)。
采用0.7%戊巴比妥钠(70 mg/kg)对小鼠进行腹腔注射麻醉并脱颈椎安乐死,剥离小鼠脑组织和空肠组织,放入福尔马林固定液中固定。进行石蜡包埋、石蜡切片、组织切片脱水和HE染色,中性树胶封片。通过全景数字扫描系统对染色结果进行扫片,并观察小鼠海马和空肠组织结构的变化。
于RFR连续暴露至28 d时,采用0.7%戊巴比妥钠(70 mg/kg)对小鼠进行麻醉,采集小鼠中央静脉血并分离血清;对小鼠脑、空肠和脾脏进行取材,加入含蛋白酶抑制剂的RIPA裂解液制作组织匀浆,在4 ℃下以12 000 r/min离心10 min,提取上清液,检测蛋白浓度。采用放射免疫法检测上述样本中促炎细胞因子(TNF-α和IL-1β)以及抑炎细胞因子(IL-4和IL-10)水平的变化。
于RFR连续暴露至28 d时,提取小鼠肠道内容物进行宏基因组测序分析。对微生物总DNA进行提取;进行文库构建;文库质检合格后进行高通量测序。统计Unigene在物种水平进行丰度分析及差异比较分析,其中,Shannon指数和Simpson指数表示不确定因素,Chao1指数和Observed species指数主要是统计群落中物种的数目,上述参数数值越大,表示物种多样性越高。
采用SPSS 26.0软件进行统计学分析。组间差异采用t检验,数据采用x¯±se表示。P<0.05表示差异具有统计学意义。
通过电磁辐射分析仪对ERC有效工作区连续监测6 min,电场强度最小值为96.1 V/m,最大值为148.6 V/m,平均值为132.5 V/m;经计算小鼠RFR全身平均SAR值为3.96 W/kg(图1A)。在本实验条件下,小鼠腹腔温度最大升幅达0.59 ℃(图1B)。上述结果提示,本实验条件下长期RFR引起小鼠核心体温升高在1 ℃以内,不会引起明显的热损伤效应。
Y-迷宫自由交替实验显示,与Con组相比,RFR组小鼠平均速度和自由交替比未见显著差异(图2A~C)。新物体识别实验显示,与Con组相比,RFR组小鼠探索新物体时间占比和辨别指数未见显著差异(图2D~F)。上述结果提示,本实验条件下长期RFR对小鼠的运动能力、空间和物体识别记忆未见明显影响。
旷场实验显示,与Con组相比,RFR组小鼠进入中心区域时间和次数未见显著差异(图3A~C)。高架十字迷宫实验显示,与Con组相比,RFR组小鼠探索开放臂时间占比和次数占比未见显著差异(图3D~F)。上述结果提示,本实验条件下长期RFR未引起小鼠产生明显焦虑行为。
糖水偏好实验显示,与Con组相比,RFR组小鼠糖水偏好时间占比和偏好指数未见明显变化(图4A、B)。悬尾实验显示,与Con组相比,RFR组小鼠不动时间占比和挣扎时间占比未见明显差异(图4C、D)。上述结果提示,在本实验条件下RFR暴露未引起小鼠产生抑郁样行为。
在RFR暴露28 d后对小鼠解剖取材,小鼠海马组织病理染色结果显示,RFR组小鼠海马呈正常组织结构,神经元细胞核大而圆,边界清晰。与Con组相比,RFR组小鼠海马组织结构未见明显损伤改变(图5A)。小鼠空肠组织病理染色结果提示,与Con组相比,RFR组小鼠空肠绒毛未见明显损伤(图5B)。上述结果提示,本实验条件下长期RFR对小鼠海马和空肠组织结构未见明显损伤效应。
放射免疫实验检测显示,与Con组相比,RFR组小鼠血清中TNF-α含量在暴露28 d后显著升高(P<0.05),在脑、脾脏、空肠组织中存在升高趋势但未见统计学差异(图6A),血清、脑、脾脏和空肠中的IL-1β、IL-4及IL-10含量均未见显著改变(图6B~D)。上述结果提示,本实验条件下长期RFR引起小鼠机体发生炎症反应。
与Con组相比,RFR组照射后小鼠菌群α多样性显著下降,主要表现为Shannon指数、Chao1指数、Simpson指数及Observed speries指数均显著下调(P<0.01,图7),提示本实验条件下RFR暴露导致小鼠肠道菌群多样性和物种丰富度显著下降。
与Con组相比,RFR组在肠道菌群种水平呈明显差异变化,主要表现为鼠乳酸杆菌(Ligilactobacillus murinus)、细菌1xD42-87(Bacterium 1xD42-87)和乙酰因子属菌(Acetatifactor muris,Acetatifactor aquisgranensis)相对丰度显著降低(P<0.05),毛螺科菌(Lachnospiraceae bacterium)相对丰度显著升高(P<0.01,图8)。综上提示,本实验条件下长期RFR引起小鼠肠道菌群多样性显著降低,肠道菌群组成发生明显变化。
在电磁辐射生物效应研究中,脑是公认的对电磁辐射敏感的靶器官之一[10]。RFR生物效应分为热效应和非热效应,热效应指RFR产热对机体带来的影响,其作用明确;而非热效应指除发热之外的其他效应,在机体发热阈值以下(≤1 ℃),RFR是否对脑功能产生不利影响尚存争议[11]。流行病学调查表明,长期处于RFR环境下的人群会出现失眠、学习记忆力下降、情绪异常、精神沉郁等症状[12]。郑荣淇等[13]通过2.65 GHz RFR暴露小鼠,4 W/kg,4 h/d,连续暴露28 d,引起小鼠的空间学习记忆下降。VARGHESE等[14]采用2.45 GHz射频电磁场对大鼠进行暴露实验,7.88 W/m2,4 h/d,连续45 d,引起大鼠记忆力下降和焦虑样行为。然而,相关研究并不统一,研究发现长期RFR对大鼠学习记忆能力和情绪行为未见显著差异[15-17]。在本部分中,RFR引起小鼠核心体温升高,最大升幅达0.59 ℃,处于热安全的范围值。在此基础上,联合多种行为学检测发现长期RFR对小鼠学习记忆能力、焦虑和抑郁情绪未见明显影响。从研究结果来看,由于脑功能具有一定的冗余和代偿能力,本实验条件下对脑功能损伤轻微,未观察到小鼠认知和情绪明显的变化。上述研究结论与所引用文献部分一致[13],在频率相同、暴露方式接近的情况下均未观察到对小鼠的焦虑和抑郁情绪造成明显影响,但有研究通过水迷宫实验发现RFR对小鼠学习记忆具有轻微的影响[13],可能因为水迷宫对于学习记忆的检测灵敏性较高,易于发现轻微的损伤变化。
炎症反应是身体对外部刺激的自我保护,表现为表观特征及功能障碍的改变,通过其自我防御和免疫调节以维持机体生理稳态。然而,过度或持续的炎症反应可能会导致组织损伤和疾病的发生。一般来说,慢性炎症促进肿瘤的发展、转移扩散及导致药物治疗耐药性的加重[18]。此外,随着时间的推移,大多数病程会从炎症性表型转变为神经退行性表型发展[19]。神经炎症涉及到免疫细胞的激活、促炎细胞因子和趋化因子的释放。例如,小胶质细胞通过活化分泌TNF-α和IL-1β等促炎细胞因子介导神经炎症。神经炎症是诱发焦虑情绪的独立风险因素之一,与杏仁核的炎性细胞因子有关[20]。大脑作为神经网络的汇聚所,RFR导致大脑产生的不利影响可能伴有炎症的发生,关于RFR如何影响大脑发生神经炎症并损伤机体等相关研究仍需深入挖掘。研究发现,1 800 MHz(SAR值为2.9 W/kg)全球移动通信系统(global system for mobile communications,GSM)单次暴露后,诱导促炎细胞因子和小胶质细胞对外周免疫应答的反应,在炎症状态下会加深对RFR的吸收并对机体造成不利影响[21]。此外,王泽泽等[22]通过电磁辐射诱发小鼠神经功能异常,致小胶质细胞被激活表现为促炎型M1表型,并发生炎症反应。有研究表明,900 MHz GSM急性暴露对中年小鼠的影响更为严重,包括情绪记忆反应的变化和皮质酮含量下降等,IL-1β的增加似乎参与着相关作用[23]。在本部分中,长期RFR可引起小鼠血清促炎细胞因子TNF-α显著上调,在脑、脾脏和空肠中有升高趋势,但未见统计学差异,提示长期RFR可引起小鼠机体发生炎症反应。此外,RFR影响小鼠血清中TNF-α升高,而脑、空肠和脾脏组织未见明显变化,可能由于RFR生物吸收剂量随着穿透深度的增加而不断降低,在本实验条件下RFR可能未引起脑、肠等深部组织器官炎症反应明显激活。此外,由于血脑屏障的存在,在一定程度上限制了血清TNF-α从循环系统向脑组织的扩散。考虑到人群接触RFR的程度不断加深,暴露时间不断延长,长期RFR导致的炎症反应对机体健康的不利影响值得关注,其产生的不良生物学后果有待进一步评估。
肠道微生物在机体炎症反应和神经免疫调控中发挥重要作用。研究发现,肠道微生态失调与情绪应激、神经退行性疾病发生密切相关[24]。肠道微生物与组织细胞之间相互作用,共同调节促炎细胞因子或趋化因子的产生,包括IL-8,IL-10和TGF-β[25]。肠道代谢物吲哚-3-乳酸可通过调节肠道微生物群来缓解肠道炎症和预防结肠炎[26]。目前,关于电磁辐射对肠道微生态的影响研究较少,机制研究不一。研究发现,4.9 GHz射频暴露后引起小鼠肠道微生物和代谢物显著差异[7]。3.5 GHz RFR诱导小鼠背侧海马CA3脑区NLRP3因子发生依赖性焦亡、肠道菌群失调及代谢紊乱,诱导小鼠产生焦虑样行为[27]。肠道微生物发生变化后,其代谢产物也会随之改变,从而产生不利影响。在本研究中,长期RFR导致小鼠肠道微生物α多样性显著下降、肠道微生物丰度和组成显著变化。在种水平上,鼠李糖乳酸杆菌和乙酰因子属菌相对丰度降低,毛螺科菌相对丰度升高。有研究表明,毛螺科菌过度生长和变形菌门菌种过度减少会影响菌群微生态,从而导致炎症性肠病的发生[28]。据此推测,肠道菌群紊乱在长期RFR介导的机体炎症反应中发挥重要作用,通过饮食调节和肠道菌群干预等可能对RFR导致健康的不利影响起到预防和保健的作用。
综上所述,2.65 GHz长期射频辐射引起小鼠机体炎症反应和肠道微生态失调,其中,肠道菌群紊乱可能是长期RFR引起机体炎症反应的重要机制。

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2025年第47卷第15期
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doi: 10.16016/j.2097-0927.202504048
  • 接收时间:2025-04-12
  • 首发时间:2026-10-28
  • 出版时间:2025-08-15
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  • 收稿日期:2025-04-12
  • 修回日期:2025-06-22
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    1佛山大学动物科技学院,广东 佛山
    2军事科学院军事医学研究院,北京

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2种不同金属材料的力学参数
科
Family
属数
Number of
genus
种数
Number of
species
占总种数比例
Percentage of
total species (%)
属
Genus
种数
Number of
species
占总种数比例
Percentage of total
species (%)
鹅膏菌科Amanitaceae 2 11 5.26 鹅膏菌属 Amanita 10 4.78
小菇科 Mycenaceae 2 12 5.74 丝盖伞属 Inocybe 5 2.39
多孔菌科 Polyporaceae 8 14 6.70 蜡蘑属 Laccaria 5 2.39
红菇科 Russulaceae 3 23 11.00 小皮伞属 Marasmius 6 2.87
小菇属 Mycena 11 5.26
光柄菇属 Pluteus 5 2.39
红菇属 Russula 17 8.13
栓菌属 Trametes 5 2.39
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