Article(id=1211268933143884234, tenantId=1146029695717560320, journalId=1189873630562394117, issueId=1211268928383348982, articleNumber=null, orderNo=null, doi=10.11855/j.issn.0577-7402.2021.02.02, pmid=null, cstr=null, oa=null, hot=null, price=null, onlineType=0, articleFormat=0, articleType=null, articleTypeStr=research-article, receivedDate=1606320000000, receivedDateStr=2020-11-26, revisedDate=1611763200000, revisedDateStr=2021-01-28, acceptedDate=null, acceptedDateStr=null, onlineDate=1766718617949, onlineDateStr=2025-12-26, pubDate=1614441600000, pubDateStr=2021-02-28, doiRegisterDate=null, doiRegisterDateStr=null, onlineIssueDate=1766718617949, onlineIssueDateStr=2025-12-26, onlineJustAcceptDate=null, onlineJustAcceptDateStr=null, onlineFirstDate=null, onlineFirstDateStr=null, sourceXml=null, magXml=null, createTime=1766718617949, creator=13701087609, updateTime=1766718617949, updator=13701087609, issue=Issue{id=1211268928383348982, tenantId=1146029695717560320, journalId=1189873630562394117, year='2021', volume='46', issue='2', pageStart='107', pageEnd='211', issueExtLink='null', onlineDate='null', pubDate='null', beforeIssueId=null, nextIssueId=null, price=null, status=1, issueComplete=1, articleOrder=1, issueType=-1, specialIssue=0, createTime=1766718616815, creator=13701087609, updateTime=1766718805938, updator=13701087609, preIssue=null, nextIssue=null, ext={EN=IssueExt(id=1211269721685627740, tenantId=1146029695717560320, journalId=1189873630562394117, issueId=1211268928383348982, language=EN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=), CN=IssueExt(id=1211269721685627741, tenantId=1146029695717560320, journalId=1189873630562394117, issueId=1211268928383348982, language=CN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=)}, issueFiles=null}, startPage=114, endPage=121, ext={EN=ArticleExt(id=1211268933483622883, articleId=1211268933143884234, tenantId=1146029695717560320, journalId=1189873630562394117, language=EN, title=Sex-specific effects of sevoflurane on cognitive function of postnatal rats, columnId=1190310110212751762, journalTitle=Medical Journal of Chinese People’s Liberation Army, columnName=Basic Research, runingTitle=null, highlight=null, articleAbstract=

Objective To investigate the gender difference of cognitive function in neonatal rats treated with sevoflurane. Methods Eighty two postnatal 7-day (P7) SD rats were randomly divided into control group and sevoflurane treated group, then divided into the following subgroups according to gender: female control group, male control group, female sevoflurane treated group, and male sevoflurane treated group. After inhaling air or 4% sevoflurane for 4 hours, the levels of serum cortisone and estradiol (E2) were detected by ELISA, and the expressions of Ki-67 and neuronal core antigen (NeuN) in hippocampus were detected by immunohistochemistry. The open field experiment was performed with rat P21 and Morris water maze experiment was performed with rat P35 to explore the effects of sevoflurane exposure on the spatial cognitive function in different gender of rats. Results ELISA reveled that, compared with the control group, the serum cortisone level in sevoflurane treated group was significantly higher [(11.60±1.41) ng/ml vs. (7.93±1.58) ng/ml, P<0.05], while the serum E2 level showed no significantly different [(35.0±8.25) pmol/L vs. (34.2±9.73) pmol/L, P>0.05]; and no significant difference existed in the levels of cortisone and E2 between different genders (P>0.05). Immunohistochemistry showed that the number of Ki-67 and NeuN positive cells in hippocampal CA1 and dentate gyrus (DG) decreased significantly in sevoflurane treated group than those in control group(P<0.05). Also, no significant difference existed in the expression of hippocampal neurons between different genders (P>0.05).Morris water maze test showed that no significant difference was found among female rats (P>0.05); compared with male control group, the platform latency and moving distance of male sevoflurane treated group were significantly prolonged at the forth and fifth day of acquired training (P<0.01 or P<0.05). At the second day of counterpoint training, the platform latency and movement distance of rats in male sevoflurane treated group were significantly prolonged (P<0.05). In counterpoint exploration training, the platform latency of rats in male sevoflurane treated group was significantly prolonged (P<0.01), and the time spent in the target quadrant reduced significantly (P=0.031). Conclusions Sevoflurane treatment has a great influence on the serum cortisone level of postnatal rats, may inhibit the proliferation of hippocampal neural stem cells and neurogenesis. The learning ability of male rats decreased after adulthood, but no obvious effect on female rats.

, correspAuthors=Meng-Meng Li, authorNote=null, correspAuthorsNote=
*E-mail:
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目的 探讨七氟烷对不同性别新生大鼠认知功能影响的差异。方法 82只出生后7 d (P7) SD大鼠,随机分为对照组与七氟烷处理组,然后按照性别分为以下亚组:雌性对照组、雄性对照组、雌性七氟烷处理组、雄性七氟烷处理组,每组19~22只。各组大鼠分别吸入空气或4%七氟烷4 h后,应用ELISA法检测外周血血清皮质酮、雌二醇(E2)水平,采用免疫组化法检测海马Ki-67和神经元核心抗原(NeuN)的表达。各组大鼠于P21行旷场实验,P35行Morris水迷宫实验。结果 ELISA检测结果显示,与对照组相比,七氟烷处理组大鼠外周血血清皮质酮水平明显升高[(7.93±1.58) ng/ml vs. (11.60±1.41) ng/ml,P<0.05],E2水平差异无统计学意义[(35.0±8.25) pmol/L vs.(34.2±9.73) pmol/L,P>0.05];不同性别各亚组间皮质酮、E2水平差异无统计学意义(P>0.05)。免疫组化检测结果显示,与对照组相比,七氟烷处理组大鼠海马CA1区和齿状回区Ki-67、NeuN阳性细胞数均明显减少(P<0.05);不同性别各亚组大鼠海马CA1区和齿状回区Ki-67、NeuN阳性细胞数差异无统计学意义(P>0.05)。Morris水迷宫实验结果显示,获得性训练第4、5天,与雄性对照组相比,雄性七氟烷处理组大鼠平台潜伏期和移动距离均明显延长(P<0.01或P<0.05),雌性对照组与雌性七氟烷处理组大鼠平台潜伏期、移动距离差异无统计学意义(P>0.05);对位训练第2天,与雄性对照组相比,雄性七氟烷处理组大鼠平台潜伏期和移动距离明显延长(P<0.05),雌性对照组与雌性七氟烷处理组大鼠平台潜伏期、移动距离差异无统计学意义(P>0.05);对位探查训练中,各处理组及不同性别大鼠游泳速度、目标象限次数比较,差异均无统计学意义(P>0.05),但与雄性对照组相比,雄性七氟烷处理组大鼠平台潜伏期明显延长(P<0.01),目标象限耗时明显缩短(P=0.031)。结论 七氟烷处理对新生大鼠血清皮质酮水平影响较大,可抑制海马神经干细胞增殖及神经元发生,成年后雄性大鼠学习能力有所减退,但对雌性大鼠无明显影响。

, correspAuthors=李萌萌, authorNote=null, correspAuthorsNote=
李萌萌,E-mail:
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武健,硕士研究生,主要从事麻醉药物对不同性别认知功能影响的研究。E-mail:

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E2. 雌二醇;与对照组比较,(1)P<0.01。

, figureFileSmall=T+yAJ+ut+ilL7xYETBT7zw==, figureFileBig=C33Ko2fs7e1dxhDTYeALHQ==, tableContent=null), ArticleFig(id=1211268943277323046, tenantId=1146029695717560320, journalId=1189873630562394117, articleId=1211268933143884234, language=EN, label=Fig.2, caption=Effects of sevoflurane exposure on hippocampal neural stem cells and mature neurons of postnatal rats, figureFileSmall=aPOVlaPDh3tB/Nfja7ONNA==, figureFileBig=UHTrOBgYgbOPL1+E5LsdNA==, tableContent=null), ArticleFig(id=1211268943373792041, tenantId=1146029695717560320, journalId=1189873630562394117, articleId=1211268933143884234, language=CN, label=图2, caption=七氟烷处理对新生大鼠海马神经干细胞和成熟神经元的影响

A. 大鼠海马CA1区和DG区Ki-67阳性细胞;B. 大鼠海马CA1区和DG区NeuN阳性细胞;与对照组比较,(1)P<0.01。

, figureFileSmall=aPOVlaPDh3tB/Nfja7ONNA==, figureFileBig=UHTrOBgYgbOPL1+E5LsdNA==, tableContent=null), ArticleFig(id=1211268943491232558, tenantId=1146029695717560320, journalId=1189873630562394117, articleId=1211268933143884234, language=EN, label=Fig.3, caption=Effects of sevoflurane exposure on the ethology of postnatal rats, figureFileSmall=dOcUHuEkwcdDStSiQEo+Og==, figureFileBig=L7TmoD98UGOBzu1vb7Z5Tg==, tableContent=null), ArticleFig(id=1211268943583507249, tenantId=1146029695717560320, journalId=1189873630562394117, articleId=1211268933143884234, language=CN, label=图3, caption=七氟烷处理对新生大鼠行为学的影响, figureFileSmall=dOcUHuEkwcdDStSiQEo+Og==, figureFileBig=L7TmoD98UGOBzu1vb7Z5Tg==, tableContent=null), ArticleFig(id=1211268943684170548, tenantId=1146029695717560320, journalId=1189873630562394117, articleId=1211268933143884234, language=EN, label=Fig.4, caption=Effects of sevoflurane exposure on the results of acquired and exploration training of postnatal rats, figureFileSmall=g0pnfj+g2c5x/0PHz1QWJQ==, figureFileBig=Xj8g5lQzJDo2rys8p9CBew==, tableContent=null), ArticleFig(id=1211268943789028152, tenantId=1146029695717560320, journalId=1189873630562394117, articleId=1211268933143884234, language=CN, label=图4, caption=七氟烷处理对新生大鼠获得性训练及探查训练结果的影响

A—B. 获得性训练;C—F. 探查训练;与雄性对照组比较,(1)P<0.01,(2)P<0.05。

, figureFileSmall=g0pnfj+g2c5x/0PHz1QWJQ==, figureFileBig=Xj8g5lQzJDo2rys8p9CBew==, tableContent=null), ArticleFig(id=1211268943885497144, tenantId=1146029695717560320, journalId=1189873630562394117, articleId=1211268933143884234, language=EN, label=Fig.5, caption=Effects of sevoflurane exposure on the results of counterpoint training, and counterpoint exploration training of postnatal rats, figureFileSmall=UIWWfKMCxHg227kgneVtbQ==, figureFileBig=klvNceYhUIBLBwX0T0IGAw==, tableContent=null), ArticleFig(id=1211268943969383224, tenantId=1146029695717560320, journalId=1189873630562394117, articleId=1211268933143884234, language=CN, label=图5, caption=七氟烷处理对新生大鼠对位训练及对位探查训练结果的影响

A—B. 对位训练;D—F. 对位探查训练;与雄性对照组比较,(1)P<0.01,(2)P<0.05。

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七氟烷对不同性别新生大鼠认知功能影响的差异分析
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武健 1, 2 , 李萌萌 2, * , 郝济伟 3 , 孟令超 2 , 张庆红 3 , 郝薏晴 1
解放军医学杂志 | 基础研究 2021,46(2): 114-121
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解放军医学杂志 | 基础研究 2021, 46(2): 114-121
七氟烷对不同性别新生大鼠认知功能影响的差异分析
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武健1, 2 , 李萌萌2, * , 郝济伟3, 孟令超2, 张庆红3, 郝薏晴1
作者信息
  • 1锦州医科大学解放军总医院第四医学中心研究生培养基地,北京 100048
  • 2解放军总医院第四医学中心麻醉科,北京 100048
  • 3全军创伤修复与组织再生重点实验室,北京 100048
  • 武健,硕士研究生,主要从事麻醉药物对不同性别认知功能影响的研究。E-mail:

通讯作者:

李萌萌,E-mail:
Sex-specific effects of sevoflurane on cognitive function of postnatal rats
Jian Wu1, 2 , Meng-Meng Li2, * , Ji-Wei Hao3, Ling-Chao Meng2, Qing-Hong Zhang3, Yi-Qing Hao1
Affiliations
  • 1Postgraduate Training Center, the Fourth Medical Center of Chinese PLA General Hospital, Jinzhou Medical University, Beijing 100048, China
  • 2Department of Anesthesiology, the Fourth Medical Center of Chinese PLA General Hospital, Beijing 100048, China
  • 3Key Laboratory of Wound Repair and Regeneration of PLA, Beijing 100048, China
出版时间: 2021-02-28 doi: 10.11855/j.issn.0577-7402.2021.02.02
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目的 探讨七氟烷对不同性别新生大鼠认知功能影响的差异。方法 82只出生后7 d (P7) SD大鼠,随机分为对照组与七氟烷处理组,然后按照性别分为以下亚组:雌性对照组、雄性对照组、雌性七氟烷处理组、雄性七氟烷处理组,每组19~22只。各组大鼠分别吸入空气或4%七氟烷4 h后,应用ELISA法检测外周血血清皮质酮、雌二醇(E2)水平,采用免疫组化法检测海马Ki-67和神经元核心抗原(NeuN)的表达。各组大鼠于P21行旷场实验,P35行Morris水迷宫实验。结果 ELISA检测结果显示,与对照组相比,七氟烷处理组大鼠外周血血清皮质酮水平明显升高[(7.93±1.58) ng/ml vs. (11.60±1.41) ng/ml,P<0.05],E2水平差异无统计学意义[(35.0±8.25) pmol/L vs.(34.2±9.73) pmol/L,P>0.05];不同性别各亚组间皮质酮、E2水平差异无统计学意义(P>0.05)。免疫组化检测结果显示,与对照组相比,七氟烷处理组大鼠海马CA1区和齿状回区Ki-67、NeuN阳性细胞数均明显减少(P<0.05);不同性别各亚组大鼠海马CA1区和齿状回区Ki-67、NeuN阳性细胞数差异无统计学意义(P>0.05)。Morris水迷宫实验结果显示,获得性训练第4、5天,与雄性对照组相比,雄性七氟烷处理组大鼠平台潜伏期和移动距离均明显延长(P<0.01或P<0.05),雌性对照组与雌性七氟烷处理组大鼠平台潜伏期、移动距离差异无统计学意义(P>0.05);对位训练第2天,与雄性对照组相比,雄性七氟烷处理组大鼠平台潜伏期和移动距离明显延长(P<0.05),雌性对照组与雌性七氟烷处理组大鼠平台潜伏期、移动距离差异无统计学意义(P>0.05);对位探查训练中,各处理组及不同性别大鼠游泳速度、目标象限次数比较,差异均无统计学意义(P>0.05),但与雄性对照组相比,雄性七氟烷处理组大鼠平台潜伏期明显延长(P<0.01),目标象限耗时明显缩短(P=0.031)。结论 七氟烷处理对新生大鼠血清皮质酮水平影响较大,可抑制海马神经干细胞增殖及神经元发生,成年后雄性大鼠学习能力有所减退,但对雌性大鼠无明显影响。

七氟烷  /  性别  /  内分泌  /  海马  /  学习记忆  /  Morris水迷宫

Objective To investigate the gender difference of cognitive function in neonatal rats treated with sevoflurane. Methods Eighty two postnatal 7-day (P7) SD rats were randomly divided into control group and sevoflurane treated group, then divided into the following subgroups according to gender: female control group, male control group, female sevoflurane treated group, and male sevoflurane treated group. After inhaling air or 4% sevoflurane for 4 hours, the levels of serum cortisone and estradiol (E2) were detected by ELISA, and the expressions of Ki-67 and neuronal core antigen (NeuN) in hippocampus were detected by immunohistochemistry. The open field experiment was performed with rat P21 and Morris water maze experiment was performed with rat P35 to explore the effects of sevoflurane exposure on the spatial cognitive function in different gender of rats. Results ELISA reveled that, compared with the control group, the serum cortisone level in sevoflurane treated group was significantly higher [(11.60±1.41) ng/ml vs. (7.93±1.58) ng/ml, P<0.05], while the serum E2 level showed no significantly different [(35.0±8.25) pmol/L vs. (34.2±9.73) pmol/L, P>0.05]; and no significant difference existed in the levels of cortisone and E2 between different genders (P>0.05). Immunohistochemistry showed that the number of Ki-67 and NeuN positive cells in hippocampal CA1 and dentate gyrus (DG) decreased significantly in sevoflurane treated group than those in control group(P<0.05). Also, no significant difference existed in the expression of hippocampal neurons between different genders (P>0.05).Morris water maze test showed that no significant difference was found among female rats (P>0.05); compared with male control group, the platform latency and moving distance of male sevoflurane treated group were significantly prolonged at the forth and fifth day of acquired training (P<0.01 or P<0.05). At the second day of counterpoint training, the platform latency and movement distance of rats in male sevoflurane treated group were significantly prolonged (P<0.05). In counterpoint exploration training, the platform latency of rats in male sevoflurane treated group was significantly prolonged (P<0.01), and the time spent in the target quadrant reduced significantly (P=0.031). Conclusions Sevoflurane treatment has a great influence on the serum cortisone level of postnatal rats, may inhibit the proliferation of hippocampal neural stem cells and neurogenesis. The learning ability of male rats decreased after adulthood, but no obvious effect on female rats.

sevoflurane  /  gender  /  endocrine  /  hippocampus  /  learning and memory  /  Morris water maze
武健, 李萌萌, 郝济伟, 孟令超, 张庆红, 郝薏晴. 七氟烷对不同性别新生大鼠认知功能影响的差异分析. 解放军医学杂志, 2021 , 46 (2) : 114 -121 . DOI: 10.11855/j.issn.0577-7402.2021.02.02
Jian Wu, Meng-Meng Li, Ji-Wei Hao, Ling-Chao Meng, Qing-Hong Zhang, Yi-Qing Hao. Sex-specific effects of sevoflurane on cognitive function of postnatal rats[J]. Medical Journal of Chinese People’s Liberation Army, 2021 , 46 (2) : 114 -121 . DOI: 10.11855/j.issn.0577-7402.2021.02.02
七氟烷具有芳香味,且不易燃、起效快、麻醉恢复快,常用于小儿全身麻醉[1]。对啮齿类动物的研究发现,麻醉剂暴露对动物的学习记忆功能可产生持久的影响,且海马神经失调在认知功能障碍的发生发展中起重要作用。既往基础研究多选用雄性动物,以避免雌孕激素水平波动干扰实验结果,但认知水平的发展具有明显的性别差异[2]。人类流行病学调查显示,早期麻醉暴露可对男性认知功能产生长期的影响,而对女性的认知功能影响较轻,其生理学机制尚待进一步研究[3-4]。新生大鼠反复给予丙泊酚,仅雄性表现出非空间记忆受损[5]。应激及性激素相关研究发现,异丙酚麻醉可导致新生大鼠体内皮质类固醇激素如皮质酮和醛固酮分泌增多[6]。Chastain-Potts等[7]研究表观遗传修饰发现,出生7 d的幼鼠暴露于七氟醚6 h可导致神经元DNA甲基化程度降低,且暴露组雌性大鼠所生雄性幼鼠Arc和JunB mRNA表达上调,具有性别差异,这些改变可能进一步导致神经发育和行为异常。然而七氟烷暴露对新生大鼠内分泌、海马神经元发生和认知功能影响的性别差异研究较少。本研究旨在探讨七氟烷对不同性别新生大鼠认知功能影响的差异。
出生7 d (P7)的SD大鼠82只,购自北京科宇动物养殖中心,随机分为对照组与七氟烷处理组,然后按照性别分为以下亚组:雌性对照组、雄性对照组、雌性七氟烷处理组、雄性七氟烷处理组,每组19~22只。采用ALC-HTP动物恒温系统维持P7幼鼠体温在(36.5±0.5) ℃。室温保持在20~25 ℃,相对湿度60%~70%,通风良好,自由摄取食物和水。本研究通过解放军总医院动物伦理委员会批准(2019-X06-11),实验过程符合国家和单位有关实验动物的管理和使用规定。
测量各组大鼠体重,然后置于一个透明有机玻璃麻醉诱导盒中,对照组自由呼吸空气4 h,七氟烷处理组吸入4%七氟烷麻醉4 h。采用实验动物麻醉机(美国MATRX公司)以6%七氟烷麻醉诱导,4%七氟烷与30%氧气混合维持麻醉4 h,期间监测幼鼠的呼吸频率,每隔15 min夹尾,确保存活,开启二氧化碳吸附器吸收废气[8-10]
自主呼吸/七氟烷暴露4 h后,各组随机取6只大鼠,断颈取血0.5 ml于抗凝管中,常温静置2 h,4 ℃下2500 r/min离心15 min,取上清于–20 ℃保存。采用ELISA试剂盒(上海酶联生物科技有限公司)检测皮质酮、E2水平,严格按照试剂盒说明书操作。
自主呼吸/七氟烷暴露4 h后,各组随机取5只大鼠,经左心室灌注生理盐水50 ml,继续灌注预冷的4%多聚甲醛100 ml,灌注完毕后断头,收集大脑标本,采用免疫组化法检测大鼠海马CA1区及齿状回(DG)区神经细胞的增殖和分化情况,具体操作步骤参照文献[11]。加入Ki-67、NeuN一抗(1:100,北京中杉金桥生物技术有限公司)和相应二抗(北京中杉金桥生物技术有限公司),采用ImageJ软件分析并计数切片中1 mm2视野内Ki-67、NeuN阳性细胞数。
大鼠(P21)在安静黑暗实验室中适应30 min后,将其放入旷场箱(长×宽×高:100 cm×100 cm×40 cm)中心位置,自由探索5 min,记录大鼠在旷场中央区及周围区所待时间。
各组大鼠(P35)进行连续15 d的Morris水迷宫实验,测试分为以下5步:(1)环境适应。大鼠在水迷宫实验室自由摄取食物和水,每天2 h,连续5 d。(2)获得性训练。水温维持在(26±1) ℃,将大鼠头朝池壁放入水中,随机取东、西、南、北4个方向之一,记录大鼠找到水下平台的时间及移动距离,以在平台上停留30 s为结束;如超过60 s,则将大鼠引导至平台,并停留30 s。每天4次,间隔15~20 min,连续训练5 d,记录大鼠平台潜伏期和移动距离等。(3)探查训练。将平台撤除,大鼠由原平台对侧象限放入池中1 min,记录大鼠逃避潜伏期时间、逃逸距离、游泳速度、原平台穿越次数、目标象限次数、目标象限耗时等。(4)对位训练。将大鼠放置于原平台所在象限的对侧象限,方法同获得性训练,连续3 d,记录大鼠平台潜伏期和移动距离等。(5)对位探查训练。方法与探查训练类似,记录大鼠逃避潜伏期时间、逃逸距离、游泳速度、原平台穿越次数、目标象限次数、目标象限耗时等。
采用SPSS 20.0软件进行统计分析。计量资料以$\bar{x}±s$表示,组间比较采用析因设计的方差分析。P<0.05为差异有统计学意义。
ELISA检测结果显示,与对照组相比,七氟烷处理组大鼠外周血血清皮质酮水平明显升高[(7.93±1.58) ng/ml vs. (11.60±1.41) ng/ml,P<0.001],E2水平无明显差异[(35.0±8.25) pmol/L vs. (34.2±9.73) pmol/L,P>0.05]。不同性别各亚组间比较,大鼠外周血血清皮质酮、E2水平差异无统计学意义(P>0.05,图1)。
免疫组化检测结果显示,与对照组相比,七氟烷处理组大鼠海马CA1区和DG区Ki-67阳性细胞数明显减少[(27.5±5.2) 个/mm2 vs. (19.3±2.9) 个/mm2;(72.5±10.6) 个/mm2 vs.(47.2±4.9) 个/mm2P<0.05],不同性别各亚组间比较差异无统计学意义(P>0.05,图2)。与对照组相比,七氟烷处理组大鼠海马CA1区和DG区NeuN阳性细胞数亦明显减少[(117.8±16.4) 个/mm2 vs.(79.4±13.1) 个/mm2;(468.5±40.5) 个/mm2 vs.(390.0±23.2) 个/mm2P<0.05],不同性别各亚组间比较差异无统计学意义(P>0.05,图2)。
旷场实验结果显示,对照组与七氟烷处理组大鼠中央区及周围区探索时间比较差异无统计学意义[中央区:(18.34±10.15) s vs. (18.91±11.38) s,P>0.05;周围区:(325.10±20.89) s vs. (335.90±11.89) s,P>0.05,图3];不同性别各亚组间比较差异无统计学意义(P>0.05,图3)。
Morris水迷宫实验结果显示,获得性训练第1、2、3天,不同处理组及不同性别大鼠平台潜伏期和移动距离比较,差异均无统计学意义(P>0.05)。获得性训练第4、5天,与雄性对照组相比,雄性七氟烷处理组大鼠平台潜伏期、移动距离均明显延长[平台潜伏期:(13.56±2.67) s vs. (22.40±7.20) s,P=0.009;(9.23±3.93) s vs. (15.18±1.70) s,P=0.01;移动距离:(348.01±69.49) cm vs. (632.23±159.43) cm,P=0.046;(290.50±68.06) cm vs. (492.03±199.80) cm,P=0.030,图4A、B];雌性对照组与雌性七氟烷处理组大鼠平台潜伏期、移动距离差异无统计学意义(P>0.05)。提示七氟烷处理组雄性大鼠随后的空间定位能力可能受到影响,但在探查训练中,各处理组及不同性别大鼠平台潜伏期、游泳速度、目标象限次数及目标象限耗时比较,差异均无统计学意义(P>0.05,图4C—F)。
对位训练第1、3天,不同处理组及不同性别大鼠平台潜伏期、移动距离比较,差异均无统计学意义(P>0.05)。第2天,与雄性对照组相比,雄性七氟烷处理组大鼠平台潜伏期、移动距离均延长[(12.19±7.52) s vs. (25.08±6.02) s,P=0.025;(343.25±113.40) cm vs.(566.87±141.81) cm,P=0.015,图5A、B];雌性对照组与雌性七氟烷处理组大鼠平台潜伏期、移动距离差异无统计学意义(P>0.05)。对位探查训练中,各处理组及不同性别大鼠游泳速度、目标象限次数比较,差异均无统计学意义(P>0.05,图5D、E);但与雄性对照组相比,雄性七氟烷处理组大鼠平台潜伏期明显延长[(12.51±2.66) s vs. (8.67±1.03) s,P<0.01,图5C],目标象限耗时明显缩短[(28.23±3.07) s vs.(19.69±4.71) s,P=0.031,图5F]。
挥发性麻醉剂被广泛应用于临床,但其作用机制及对神经系统发育的影响尚不明确。研究发现,发育期麻醉药物暴露对大鼠随后的神经系统发育具有一定的影响,且存在性别差异[8-9],其生理机制有待进一步研究。Xie等[14]发现,七氟醚麻醉可损害成年大鼠的短期认知功能,且对雄性大鼠的损害较雌性重,其机制可能是通过下调磷酸化cAMP反应元件结合蛋白1(p-CREB1)、Bcl-2的表达,上调caspase-8的表达,从而促进海马神经元凋亡。本研究发现,七氟烷处理明显增高了大鼠外周血血清皮质酮水平,但对E2水平无明显影响;七氟烷处理可抑制海马神经干细胞的增殖及海马神经元发生,但无性别差异;七氟烷处理对雄性大鼠的空间认知功能有一定影响,但对雌性大鼠无明显影响。
神经甾体是由脑组织细胞合成的甾体激素,可能参与调控γ-氨基丁酸A型受体(gamma-aminobutyric acid type A receptor,GABAAR)的活性并对神经系统发育产生影响[15-16]。研究发现,七氟烷可通过激活边缘系统-下丘脑-垂体-肾上腺轴(limbic-hypothalamo pituitary adrenal axis,LHPA轴)引起神经活性甾体释放[17]。Xu等[18]发现,七氟烷暴露6 h可导致新生大鼠皮质酮水平增高。早期应激如长期或反复的母子分离,可导致后期严重的行为异常,以及对应激表现出更强烈的内分泌反应,成年雄性啮齿动物可能表现出更明显的异常[19]。E2是神经活性甾体激素,在发育期脑内维持于较高水平,大脑皮质中含量无性别差异,但雄性大鼠下丘脑中含量明显高于雌性[20-21]。孕晚期或出生后早期大鼠给予外源性E2,可降低成年后雌性大鼠的学习记忆功能[22]。本研究结果显示,七氟烷处理可明显增高新生大鼠血清皮质酮水平,与既往研究结果一致[18],但未发现性别差异;七氟烷处理对新生大鼠血清E2水平无明显影响,分析原因为麻醉引起的E2水平升高主要发生在脑内,但由于实验条件限制,本研究未测定各组大鼠麻醉后脑内神经活性甾体激素的水平,因此,七氟烷处理对发育期大鼠E2的影响是目前研究的重要局限。
神经元发生是神经干细胞不断增殖、分裂产生新神经元的过程。全身麻醉药物引起的大鼠认知功能障碍主要表现为海马控制的空间定向力受损及学习记忆功能下降,但其发生机制尚未明确[23]。有研究发现,海马颗粒下区齿状回神经干细胞产生发育成熟神经元的能力是决定海马功能的重要因素[24]。脓毒症小鼠海马齿状回神经干细胞分化减少,成熟神经元凋亡增加,学习和空间记忆功能随之受损[25]。Ki-67是一种细胞增殖相关核抗原,其功能与细胞有丝分裂密切相关,是目前判断细胞增殖能力的主要指标之一。NeuN是成熟神经元的特异性标志物。景灵等[26]使用七氟烷处理老年大鼠,发现大鼠海马神经干细胞(Ki-67标记)、成熟神经元(NeuN标记)和星形胶质细胞(GFAP标记)明显减少,出现神经发生失衡现象。本研究结果显示,吸入4%七氟烷可导致新生大鼠海马Ki-67、NeuN阳性细胞明显减少,神经干细胞增殖受损。新生期海马神经干细胞作为海马神经元发生的起始点,其数量减少对海马正常发育可能造成一定的影响,且可能影响远期的神经功能。
有研究发现,多次异氟烷处理与出生后啮齿类动物大脑神经元凋亡和认知功能障碍相关[27]。与雌性大鼠相比,雄性大鼠对麻醉药(异氟醚或苯巴比妥)的神经毒性作用更敏感,主要表现为海马神经元数量明显减少[28]。Rothstein等[29]认为,突触形成早期阶段(出生后即刻)全麻暴露大鼠的认知能力严重受损,且对成年期(1.5个月)雄性大鼠的影响较雌性大鼠重。脑源性神经营养因子(brain-derived neurotrophic factor,BDNF)是一种分泌性蛋白质,参与调控神经元的生长和发育,且与突触形成有关,而雌激素已被证实能增加BDNF的表达[30],这可能是麻醉药物对雌性影响较小的原因。目前Morris水迷宫为测试大鼠学习记忆功能的经典模型,主要用于客观评价大鼠空间认知功能的行为学研究。为排除七氟烷处理可能引起新生大鼠随后的精神焦虑、探索能力下降等自主行为改变,本研究在Morris水迷宫实验前,进行了旷场实验筛查,结果显示,出生后经历七氟烷处理的大鼠,其精神状况、探索及自主行为等无异常改变。Morris水迷宫实验中,七氟烷处理的雄性大鼠在获得性训练和对位训练中的平台潜伏期及移动距离均延长,提示七氟烷处理可能仅影响新生大鼠学习过程中的表现,而对随后的空间认知能力无明显影响,与既往研究结果一致[31];同时在对位探查训练中平台潜伏期及非目标象限耗时均延长,提示七氟烷处理对新生雄性大鼠的工作记忆能力有一定损害。由此可见,七氟烷处理对新生雄性大鼠随后的认知功能有一定影响,但在雌性大鼠的行为学研究中未发现类似结果。
麻醉剂不仅影响细胞凋亡,还可影响神经炎症[32]、神经元信号变化、细胞增殖及突触形成[33-34],但这些影响可通过雌激素和孕酮激素的保护作用而减轻[35]。17β-雌二醇可通过激活PI3K/Akt/Bcl-2信号通路对氯胺酮诱导的神经元凋亡发挥神经保护作用[36]。但本研究中,七氟烷处理对新生大鼠血清皮质酮和E2的影响不能解释行为学上的性别差异。有研究发现,异氟醚处理的雄性大鼠与雌性大鼠海马和丘脑核内神经元死亡情况相似[37]。Ju等[38]发现,大鼠暴露于七氟醚,其后代在行为测试和钾离子-氯离子共转运体2(KCC2)表达方面出现性别差异,仅雄性表现异常。由于激素水平的差异以及麻醉对神经系统发育可能产生的影响,雌性大鼠的恢复及适应能力可能更具弹性。
综上所述,本研究结果显示,七氟烷处理对新生大鼠血清皮质酮水平影响较大,且可抑制海马神经干细胞增殖及神经元发生,成年后雄性大鼠的学习能力有所减退,但对雌性大鼠无明显影响。麻醉剂对不同性别大鼠生理及学习认知行为的影响仍有待进一步探讨。
  • 国家自然科学基金(81272030)
  • 吴阶平医学基金会临床科研专项资助基金(320.6750.13220)
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2021年第46卷第2期
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doi: 10.11855/j.issn.0577-7402.2021.02.02
  • 接收时间:2020-11-26
  • 首发时间:2025-12-26
  • 出版时间:2021-02-28
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  • 收稿日期:2020-11-26
  • 修回日期:2021-01-28
基金
National Natural Science Foundation of China(81272030)
国家自然科学基金(81272030)
Clinical Research Special Funds of Wu Jie-Ping Medical Foundation(320.6750.13220)
吴阶平医学基金会临床科研专项资助基金(320.6750.13220)
作者信息
    1锦州医科大学解放军总医院第四医学中心研究生培养基地,北京 100048
    2解放军总医院第四医学中心麻醉科,北京 100048
    3全军创伤修复与组织再生重点实验室,北京 100048

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2种不同金属材料的力学参数

Family
属数
Number of
genus
种数
Number of
species
占总种数比例
Percentage of
total species (%)

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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