Article(id=1190310109587800463, tenantId=1146029695717560320, journalId=1189873630562394117, issueId=1190243275249390089, articleNumber=null, orderNo=null, doi=10.11855/j.issn.0577-7402.0804.2025.0218, pmid=null, cstr=null, oa=null, hot=1, price=null, onlineType=0, articleFormat=0, articleType=null, articleTypeStr=research-article, receivedDate=1717689600000, receivedDateStr=2024-06-07, revisedDate=null, revisedDateStr=null, acceptedDate=1722355200000, acceptedDateStr=2024-07-31, onlineDate=1761721645017, onlineDateStr=2025-10-29, pubDate=1748361600000, pubDateStr=2025-05-28, doiRegisterDate=null, doiRegisterDateStr=null, onlineIssueDate=1761721645017, onlineIssueDateStr=2025-10-29, onlineJustAcceptDate=null, onlineJustAcceptDateStr=null, onlineFirstDate=null, onlineFirstDateStr=null, sourceXml=null, magXml=null, createTime=1761721645017, creator=13701087609, updateTime=1769158569818, updator=13701087609, issue=Issue{id=1190243275249390089, tenantId=1146029695717560320, journalId=1189873630562394117, year='2025', volume='50', issue='5', pageStart='505', pageEnd='640', issueExtLink='null', onlineDate='null', pubDate='null', beforeIssueId=null, nextIssueId=null, price=null, status=1, issueComplete=1, articleOrder=1, issueType=-1, specialIssue=null, createTime=1761705710470, creator=13701087609, updateTime=1765784077922, updator=13701087609, preIssue=null, nextIssue=null, ext={EN=IssueExt(id=1207349188233372409, tenantId=1146029695717560320, journalId=1189873630562394117, issueId=1190243275249390089, language=EN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=), CN=IssueExt(id=1207349188233372410, tenantId=1146029695717560320, journalId=1189873630562394117, issueId=1190243275249390089, language=CN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=)}, issueFiles=null}, startPage=575, endPage=580, ext={EN=ArticleExt(id=1190310110296637844, articleId=1190310109587800463, tenantId=1146029695717560320, journalId=1189873630562394117, language=EN, title=Effects of advanced maternal age on hippocampal synaptic pruning in offspring rats, columnId=1190310110212751762, journalTitle=Medical Journal of Chinese People’s Liberation Army, columnName=Basic Research, runingTitle=null, highlight=null, articleAbstract=

Objective To investigate the effects of advanced maternal age (AMA) on synaptic pruning of the hippocampus of offspring rats. Methods Offspring of 3-month-old SD female rats and 3-month-old SD male rats were designated as the appropriate age group (Ctl group), while offspring of 12-month-old SD female rats and 3-month-old SD male rats were assigned to AMA group. Hippocampal tissues from both groups (n=3 per group at each time point) were subjected to Golgi staining on postnatal days (P) 7, 28, and 60. Dendritic complexity, spine density (DS), and morphology of immature brain neurons at different age stages were compared between the two groups. Results Compared with Ctl group, the dendritic length of AMA group on P7, P28, and P60 was significantly shortened, and the average number of intersections between dendrites and concentric circles on P28 and P60 was also significantly decreased (P<0.05). Compared with Ctl group, the dendritic spine density of AMA group was significantly decreased on P7, P28, and P60. Notably, the density of thin-shaped dendritic spines in AMA group was increased on P7, whereas the densities of stubby-shaped and mushroom-shaped dendritic spines were decreased (P<0.05). On P28 in AMA group, the densities of thin-shaped and mushroom-shaped dendritic spines were significantly increased compared with Ctl group, while the density of stubby-shaped dendritic spines was significantly decreased (P<0.05). The densities of stubby-shaped and mushroom-shaped dendritic spine were markedly lower in AMA group on P60 than those in Ctl group (P<0.05). Conclusions During postnatal brain development periods, there were abnormalities in synaptic pruning in the hippocampal region of the offspring of the AMA, which manifested as reduced dendritic complexity, decreased dendritic spine density, and disorganized dendritic spine morphology. It could be one of the key mechanisms underlying brain dysfunction in the offspring of AMA.

, correspAuthors=Li Jiang, authorNote=null, correspAuthorsNote=
E-mail:
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目的 探讨高龄妊娠对子代大鼠海马突触修剪的影响。方法 将3月龄SD雌鼠与3月龄SD雄鼠合笼所生子代记为适龄组(Ctl组),12月龄SD雌鼠与3月龄SD雄鼠合笼所生子代记为高龄组(AMA组),取两组生后第7、28、60天(每时间点每组3只)的海马组织进行高尔基染色,比较两组各年龄段未成熟脑神经元的树突复杂性、树突棘密度及形态变化。结果 与Ctl组比较,AMA组生后第7、28、60天的树突长度明显缩短,第28、60天的树突与同心圆平均交点数亦明显减少(P<0.05)。与Ctl组比较,AMA组生后第7、28、60天树突棘密度均明显降低,其中生后第7天细长型树突棘密度明显增高,而短粗型、蘑菇型树突棘密度明显降低(P<0.05),生后第28天细长型、蘑菇型树突棘密度明显增高,而短粗型树突棘密度明显降低(P<0.05),生后第60天短粗型、蘑菇型树突棘密度均明显降低(P<0.05)。结论 在高龄妊娠子代出生后脑发育过程中,海马区存在突触修剪异常,表现为树突复杂性降低,树突棘密度降低、形态紊乱,这可能是高龄妊娠子代脑功能障碍的机制之一。

, correspAuthors=蒋莉, authorNote=null, correspAuthorsNote=
蒋莉,E-mail:
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Front Synaptic Neurosci, 2014, 6: 5., articleTitle=The BDNF effects on dendritic spines of mature hippocampal neurons depend on neuronal activity, refAbstract=null), Reference(id=1190330534145393567, tenantId=1146029695717560320, journalId=1189873630562394117, articleId=1190310109587800463, doi=null, pmid=null, pmcid=null, year=2017, volume=72, issue=11, pageStart=1465, pageEnd=1473, url=null, language=null, rfNumber=[32], rfOrder=31, authorNames=Sampino S, Stankiewicz AM, Zacchini F, journalName=J Gerontol A Biol Sci Med Sci, refType=null, unstructuredReference=Sampino S, Stankiewicz AM, Zacchini F, et al. Pregnancy at advanced maternal age affects behavior and hippocampal gene expression in mouse offspring[J]. J Gerontol A Biol Sci Med Sci, 2017, 72(11): 1465-1473., articleTitle=Pregnancy at advanced maternal age affects behavior and hippocampal gene expression in mouse offspring, refAbstract=null), Reference(id=1190330534216696736, tenantId=1146029695717560320, journalId=1189873630562394117, articleId=1190310109587800463, doi=null, pmid=null, pmcid=null, year=2021, volume=46, issue=5, pageStart=456, pageEnd=461, url=null, language=null, rfNumber=[33], rfOrder=32, authorNames=潘亚男, 韩慰, 杨静, journalName=解放军医学杂志, refType=null, unstructuredReference=潘亚男, 韩慰, 杨静, 等. 高龄妊娠对子代大鼠海马甲基化水平的影响[J]. 解放军医学杂志, 2021, 46(5): 456-461., articleTitle=高龄妊娠对子代大鼠海马甲基化水平的影响, refAbstract=null)], funds=[Fund(id=1190330530571846526, tenantId=1146029695717560320, journalId=1189873630562394117, articleId=1190310109587800463, awardId=82301654, language=EN, fundingSource=National Natural Science Foundation of China(82301654), fundOrder=null, country=null), Fund(id=1190330530655732607, tenantId=1146029695717560320, journalId=1189873630562394117, articleId=1190310109587800463, awardId=KJQN202100423, language=EN, fundingSource=Scientific and Technological Research Program of Chongqing Municipal Education Commission(KJQN202100423), fundOrder=null, country=null)], companyList=[AuthorCompany(id=1190330527619056443, tenantId=1146029695717560320, journalId=1189873630562394117, articleId=1190310109587800463, xref=null, ext=[AuthorCompanyExt(id=1190330527631639356, tenantId=1146029695717560320, journalId=1189873630562394117, articleId=1190310109587800463, companyId=1190330527619056443, language=EN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=Department of Neurology, Children's Hospital of Chongqing Medical University/National Clinical Research Center for Child Health and Disorders/Ministry of Education Key Laboratory of Child Development and Disorders/Chongqing Key Laboratory of Child Neurodevelopment and Cognitive Disorders, Chongqing 400014, China), AuthorCompanyExt(id=1190330527640027965, tenantId=1146029695717560320, journalId=1189873630562394117, articleId=1190310109587800463, companyId=1190330527619056443, language=CN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=重庆医科大学附属儿童医院神经内科/国家儿童健康与疾病临床医学研究中心/儿童发育疾病研究教育部重点实验室/ 儿童神经发育与认知障碍重庆市重点实验室,重庆 400014)])], figs=[ArticleFig(id=1190330529896563574, tenantId=1146029695717560320, journalId=1189873630562394117, articleId=1190310109587800463, language=EN, label=Fig.1, caption=Processes for analyzing dendritic complexity, figureFileSmall=HYxZhVHeUJAOYVKJdYNrQg==, figureFileBig=fTv4xZOn4kpi8MFvi07t8g==, tableContent=null), ArticleFig(id=1190330530014004087, tenantId=1146029695717560320, journalId=1189873630562394117, articleId=1190310109587800463, language=CN, label=图1, caption=树突复杂性的分析流程

A. 高尔基染色后海马区神经元图像;B. 追踪目标神经元树突轨迹以获得树突长度;C. 将轨迹图转换为二进制格式获得目标神经元树突二维分布图;D. 以目标神经元胞体为中心进行Sholl分析

, figureFileSmall=HYxZhVHeUJAOYVKJdYNrQg==, figureFileBig=fTv4xZOn4kpi8MFvi07t8g==, tableContent=null), ArticleFig(id=1190330530123055992, tenantId=1146029695717560320, journalId=1189873630562394117, articleId=1190310109587800463, language=EN, label=Fig.2, caption=Diagram of dendritic spine morphology classification in hippocampal neurons, figureFileSmall=HqV/7hT746d6DRMAkXYxsg==, figureFileBig=HueGEYIW3CBRkeFRRnwjmA==, tableContent=null), ArticleFig(id=1190330530181776249, tenantId=1146029695717560320, journalId=1189873630562394117, articleId=1190310109587800463, language=CN, label=图2, caption=海马区神经元树突棘形态分类示意图, figureFileSmall=HqV/7hT746d6DRMAkXYxsg==, figureFileBig=HueGEYIW3CBRkeFRRnwjmA==, tableContent=null), ArticleFig(id=1190330530236302202, tenantId=1146029695717560320, journalId=1189873630562394117, articleId=1190310109587800463, language=EN, label=Fig.3, caption=Comparison of dendritic complexity of hippocampus neurons in offspring rats between the two groups, figureFileSmall=esNFUfqq09MNzuudnXGu0Q==, figureFileBig=f8wzWOGVSjUwgNWJV/2EQg==, tableContent=null), ArticleFig(id=1190330530315993979, tenantId=1146029695717560320, journalId=1189873630562394117, articleId=1190310109587800463, language=CN, label=图3, caption=两组大鼠海马区神经元树突复杂性比较

Ctl组. 适龄组;AMA组. 高龄组;A. 生后第7、28、60天两组大鼠海马神经元树突与同心圆交点分布,相邻两个同心圆间隔10 μm;B. 生后第7、28、60天两组大鼠海马树突长度及树突与同心圆平均交点数的柱状图;纳入统计的神经元个数:第7天,Ctl组56个、AMA组58个;第28天,Ctl组76个、AMA组85个;第60天,Ctl组102个、AMA组104个;与Ctl组比较,*P<0.05,**P<0.01,***P<0.001

, figureFileSmall=esNFUfqq09MNzuudnXGu0Q==, figureFileBig=f8wzWOGVSjUwgNWJV/2EQg==, tableContent=null), ArticleFig(id=1190330530387297148, tenantId=1146029695717560320, journalId=1189873630562394117, articleId=1190310109587800463, language=EN, label=Fig.4, caption=Effect of advanced maternal age on dendritic spines of hippocampus in offspring rats, figureFileSmall=JDzXwdVKez8iveuZfOEixQ==, figureFileBig=l9MyfatN69cn+B4iU6exeA==, tableContent=null), ArticleFig(id=1190330530454406013, tenantId=1146029695717560320, journalId=1189873630562394117, articleId=1190310109587800463, language=CN, label=图4, caption=高龄妊娠对子代大鼠海马区树突棘的影响

Ctl组. 适龄组;AMA组. 高龄组;A、B、C. 生后第7、28、60天两组海马区不同形态树突棘密度;纳入统计的树突节段的段数:第7天,Ctl组129个、AMA组146个;第28天,Ctl组235个、AMA组225个;第60天,Ctl组188个、AMA组213个;与Ctl组比较,*P<0.05,**P<0.01,***P<0.001

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高龄妊娠对子代大鼠海马突触修剪的影响
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蒋春雪 , 韩子瑶 , 严心汝 , 韩慰 , 蒋莉 *
解放军医学杂志 | 基础研究 2025,50(5): 575-580
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解放军医学杂志 | 基础研究 2025, 50(5): 575-580
高龄妊娠对子代大鼠海马突触修剪的影响
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蒋春雪, 韩子瑶, 严心汝, 韩慰, 蒋莉*
作者信息
  • 重庆医科大学附属儿童医院神经内科/国家儿童健康与疾病临床医学研究中心/儿童发育疾病研究教育部重点实验室/ 儿童神经发育与认知障碍重庆市重点实验室,重庆 400014

通讯作者:

蒋莉,E-mail:
Effects of advanced maternal age on hippocampal synaptic pruning in offspring rats
Chun-Xue Jiang, Zi-Yao Han, Xin-Ru Yan, Wei Han, Li Jiang*
Affiliations
  • Department of Neurology, Children's Hospital of Chongqing Medical University/National Clinical Research Center for Child Health and Disorders/Ministry of Education Key Laboratory of Child Development and Disorders/Chongqing Key Laboratory of Child Neurodevelopment and Cognitive Disorders, Chongqing 400014, China
出版时间: 2025-05-28 doi: 10.11855/j.issn.0577-7402.0804.2025.0218
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目的 探讨高龄妊娠对子代大鼠海马突触修剪的影响。方法 将3月龄SD雌鼠与3月龄SD雄鼠合笼所生子代记为适龄组(Ctl组),12月龄SD雌鼠与3月龄SD雄鼠合笼所生子代记为高龄组(AMA组),取两组生后第7、28、60天(每时间点每组3只)的海马组织进行高尔基染色,比较两组各年龄段未成熟脑神经元的树突复杂性、树突棘密度及形态变化。结果 与Ctl组比较,AMA组生后第7、28、60天的树突长度明显缩短,第28、60天的树突与同心圆平均交点数亦明显减少(P<0.05)。与Ctl组比较,AMA组生后第7、28、60天树突棘密度均明显降低,其中生后第7天细长型树突棘密度明显增高,而短粗型、蘑菇型树突棘密度明显降低(P<0.05),生后第28天细长型、蘑菇型树突棘密度明显增高,而短粗型树突棘密度明显降低(P<0.05),生后第60天短粗型、蘑菇型树突棘密度均明显降低(P<0.05)。结论 在高龄妊娠子代出生后脑发育过程中,海马区存在突触修剪异常,表现为树突复杂性降低,树突棘密度降低、形态紊乱,这可能是高龄妊娠子代脑功能障碍的机制之一。

高龄妊娠  /  子代大鼠  /  海马  /  树突  /  神经元可塑性

Objective To investigate the effects of advanced maternal age (AMA) on synaptic pruning of the hippocampus of offspring rats. Methods Offspring of 3-month-old SD female rats and 3-month-old SD male rats were designated as the appropriate age group (Ctl group), while offspring of 12-month-old SD female rats and 3-month-old SD male rats were assigned to AMA group. Hippocampal tissues from both groups (n=3 per group at each time point) were subjected to Golgi staining on postnatal days (P) 7, 28, and 60. Dendritic complexity, spine density (DS), and morphology of immature brain neurons at different age stages were compared between the two groups. Results Compared with Ctl group, the dendritic length of AMA group on P7, P28, and P60 was significantly shortened, and the average number of intersections between dendrites and concentric circles on P28 and P60 was also significantly decreased (P<0.05). Compared with Ctl group, the dendritic spine density of AMA group was significantly decreased on P7, P28, and P60. Notably, the density of thin-shaped dendritic spines in AMA group was increased on P7, whereas the densities of stubby-shaped and mushroom-shaped dendritic spines were decreased (P<0.05). On P28 in AMA group, the densities of thin-shaped and mushroom-shaped dendritic spines were significantly increased compared with Ctl group, while the density of stubby-shaped dendritic spines was significantly decreased (P<0.05). The densities of stubby-shaped and mushroom-shaped dendritic spine were markedly lower in AMA group on P60 than those in Ctl group (P<0.05). Conclusions During postnatal brain development periods, there were abnormalities in synaptic pruning in the hippocampal region of the offspring of the AMA, which manifested as reduced dendritic complexity, decreased dendritic spine density, and disorganized dendritic spine morphology. It could be one of the key mechanisms underlying brain dysfunction in the offspring of AMA.

advanced maternal age  /  offspring rat  /  hippocampus  /  dendrites  /  neuronal plasticity
蒋春雪, 韩子瑶, 严心汝, 韩慰, 蒋莉. 高龄妊娠对子代大鼠海马突触修剪的影响. 解放军医学杂志, 2025 , 50 (5) : 575 -580 . DOI: 10.11855/j.issn.0577-7402.0804.2025.0218
Chun-Xue Jiang, Zi-Yao Han, Xin-Ru Yan, Wei Han, Li Jiang. Effects of advanced maternal age on hippocampal synaptic pruning in offspring rats[J]. Medical Journal of Chinese People’s Liberation Army, 2025 , 50 (5) : 575 -580 . DOI: 10.11855/j.issn.0577-7402.0804.2025.0218
高龄产妇是指分娩年龄≥35岁的妇女。随着辅助生殖技术的进步、女性受教育程度的提高及优先考虑就业等社会因素的影响,女性生育年龄不断延迟,全球各地高龄生育的现象呈上升趋势[1-2]。高龄产妇卵母细胞质量下降及胚胎着床受损直接影响胚胎质量和子代发育[3-4]。高龄妊娠被认为是各种妊娠并发症及不良妊娠结局的危险因素,尤其是对子代神经发育的负面影响越来越受到关注[5]。有研究发现,高龄妊娠子代罹患焦虑、抑郁、孤独症谱系障碍等脑功能障碍的风险更高[6-7],但目前相关机制研究尚少。树突、轴突及突触是神经元之间通信的重要结构。树突主要负责接收神经元的传入信号,这些信号经胞体处理后沿着轴突传播至其他神经元的树突或胞体,其中轴突末端与其他神经元胞体或树突形成的连接为突触。突触修剪是清除“无意义”突触的过程,是突触可塑性的基础,对大脑形成正确的神经环路及认知功能至关重要。本课题组前期研究发现,生后早期高龄妊娠子代海马存在突触功能障碍,包括突触后致密物-95及突触素表达降低,突触后致密物厚度减小,长时程增强存在缺陷[8],而突触可塑性是否受突触修剪的影响尚不明确。本研究通过构建高龄妊娠子代大鼠模型,探究高龄妊娠对子代海马突触修剪的影响,旨在为探讨高龄妊娠导致子代脑功能障碍的机制提供新的思路。
SPF级3月龄(n=6)、12月龄(n=6)雌性SD大鼠及3月龄(n=6)雄性SD大鼠购自重庆医科大学动物实验中心[实验动物生产许可证号:SYXK(渝)2022-0002],在SPF级环境中以标准饲料喂养,自由摄食水,昼夜周期为12 h/12 h,环境温度19~25 ℃,相对湿度50%~60%。冰冻切片机购自德国Leica公司,荧光显微镜购自日本尼康公司,快速Golgi-Cox染色试剂盒购自美国FD Neuro Technologies公司,O.C.T冰冻切片包埋剂购自日本樱花公司。本研究获重庆医科大学附属儿童医院实验动物伦理委员会审批(CHCMU-IACUC20240131011)。
3月龄及12月龄雌鼠分别与3月龄雄鼠按照1:1的比例交配,产下的子代分别记为适龄组(control,Ctl组)和高龄组(advanced maternal aged,AMA组);生后第7、28、60天时两组各随机选取3只大鼠,收集全脑用于后续实验检测及分析。
以3%戊巴比妥钠溶液腹腔注射麻醉大鼠,分离脑组织,参照快速Golgi-Cox染色试剂盒说明书进行高尔基染色。脑组织于浸渍液中浸泡2周后进行包埋、冠状位切片。
光学显微镜白光下采用4倍及10倍物镜观察海马区神经元分布,选取海马区神经元结构清晰、神经元之间交集重叠较少、神经元尽量完整的脑组织切片拍照。采用100倍油浸物镜观察海马区神经元树突棘数量及形态,Z轴堆叠技术扫描(Z步长:2 μm)脑组织切片(厚度:120 μm),反卷积技术合成三维图像。
在大脑两侧海马CA1、CA3及齿状回(DG)区中分别随机选取3~6个神经元图像进行树突复杂性、树突棘密度及形态分析。
树突复杂性是指神经元树突长度及树突分枝数的情况。Sholl分析法是以神经元胞体为圆心,以10 μm递增的半径画同心圆,统计树突平均长度、树突与同心圆平均交点数,从而间接反映树突的复杂性。采用ImageJ软件测量树突长度,获得神经二维分布图,使用Sholl分析法插件分析树突与同心圆交点数(图1)。
树突棘密度以每30 μm树突长度中树突棘的数量来表示。使用Case Viewer软件选取二级或三级树突节段进行分析。纳入分析的树突节段需要同时满足以下3个标准:(1)树突长度30~50 μm且未截断;(2)可与附近神经元区分;(3)树突颜色深度均匀一致[9]。树突棘形态分为以下5种:(1)丝状伪足型,细长无颈部,长度>2 μm;(2)细长型,颈部长而细,头部小而圆,颈部长度<2 μm;(3)短粗型,无明显颈部,头部长度与宽度的比约为1:1;(4)蘑菇型,颈部短而粗,头部宽度>0.6 μm;(5)分枝型,头部2个或以上分支(图2)[10]
采用GraphPad Prism 9.0软件对数据进行统计分析及制图。计量资料均符合正态分布,以x±s表示,组间比较采用独立样本t检验。P<0.05为差异有统计学意义。
生后第7、28、60天,AMA组海马神经元树突长度较Ctl组均明显缩短[(30.53±1.37) μm vs. (38.26±1.14) μm,(49.94±1.80) μm vs. (67.98±2.17) μm,(59.30±1.54) μm vs. (64.77±1.46) μm;P<0.001或P<0.05,图3A];生后第28、60天,AMA组海马神经元树突与同心圆平均交点数均较Ctl组明显减少[(33.55±1.61)个 vs. (41.78±2.77)个,(33.49±1.38)个 vs. (44.41±1.42)个;P<0.01或P<0.001,图3A、B]。
生后第7、28、60天AMA组海马神经元的树突棘密度较Ctl组均明显降低[(11.06±0.35)个/30 μm vs. (12.71±0.37)个/30 μm,(15.17±0.26)个/30 μm vs. (17.90±0.28)个/30 μm,(15.94±0.27)个/30 μm vs. (20.55±0.34)个/30 μm;P<0.01或P<0.001]。
生后第7天,与Ctl组比较,AMA组海马神经元短粗型、蘑菇型树突棘密度明显降低[(6.79±0.21)个/30 μm vs. (9.05±0.24)个/30 μm,(0.35±0.06)个/30 μm vs. (0.54±0.07)个/30 μm;P<0.001或P<0.05],细长型树突棘密度明显增高[(3.07±0.21)个/30 μm vs. (2.36±0.18)个/30 μm;P<0.05],丝状伪足型、分枝型树突棘密度虽有增高趋势,但差异无统计学意义[(0.72±0.08)个/30 μm vs. (0.64±0.09)个/30 μm,(0.12±0.03)个/30 μm vs. (0.12±0.03)个/30 μm;P>0.05](图4A)。
生后第28天,与Ctl组比较,AMA组海马神经元细长型、蘑菇型树突棘密度明显增高[(2.69±0.12)个/30 μm vs. (1.87±0.10)个/30 μm,(0.67±0.05)个/30 μm vs. (0.49±0.04)个/30 μm,P<0.001或P<0.01],而短粗型树突棘密度明显降低[(15.41±0.25)个/30 μm vs. (11.66±0.21)个/30 μm,P<0.001],丝状伪足型、分枝型树突棘密度有增高趋势,但差异无统计学意义[(0.11±0.02)个/30 μm vs. (0.09±0.02)个/30 μm,(0.04±0.02)个/30 μm vs. (0.04±0.01)个/30 μm;P>0.05](图4B)。
生后第60天,与Ctl组比较,AMA组海马神经元短粗型、蘑菇型树突棘密度明显降低[(11.97±0.23)个/30 μm vs. (16.09±0.29)个/30 μm,(1.37±0.08)个/30 μm vs. (1.90±0.11)个/30 μm;P<0.001],细长型树突棘密度有增高趋势,丝状伪足型、分枝型树突棘密度有降低趋势,但差异均无统计学意义[(2.48±0.11)个/30 μm vs. (2.42±0.11)个/30 μm,(0.09±0.02)个/30 μm vs. (0.10±0.02)个/30 μm,(0.03±0.01)个/30 μm vs. (0.04±0.01)个/30 μm;P>0.05](图4C)。
高龄妊娠在全球呈逐年增加趋势[11]。1992-2018年,加拿大高龄产妇比例由9.9%上升至23.5%[12];2009-2019年,韩国高龄产妇比例由15.4%上升至33.4%[13];随着中国三孩生育政策的开放,2013-2022年,北京高龄产妇比例由11.4%上升至34.5%[14]。临床研究发现,高龄产妇子代更易发生双相情感障碍、精神分裂症等神经精神类疾病[15]。本课题组前期研究发现,与适龄妊娠大鼠子代相比,高龄妊娠大鼠子代在新物体识别实验及Morris水迷宫实验中表现出学习和记忆能力的明显下降,在高架十字迷宫实验及强迫游泳实验中表现出一定的焦虑、抑郁倾向[16-17],但具体机制不明。高龄妊娠子代脑发育相关问题日益突出,故探索其潜在发病机制、寻找有效治疗策略是小儿神经及康复领域的研究热点。
在人类婴幼儿时期突触发展到最高峰时,中枢神经系统形成大量树突和轴突,脑内突触会经历爆发式增长,导致神经元之间形成过量的突触连接,每个神经元大约有15 000个突触,这一数量接近成人的2倍。突触连接的数量受突触修剪调控,在脑发育过程中突触会发生依赖于活动的重新排列,“多余”突触将被清除,若该进程受到干扰可能导致突触结构和功能异常[18]。在精神分裂症、阿尔茨海默病等神经精神类疾病中均存在海马区突触修剪异常,目前认为可能是突触修剪异常破坏了兴奋性突触与抑制性突触之间的平衡,并在上述疾病的发生中扮演了重要角色[19]
树突长度和树突棘形态反映了突触修剪的结果。有研究发现,精神分裂症和双相情感障碍患者均存在特定皮质区的树突长度缩短及树突棘丢失,可能与星形胶质细胞及小胶质细胞的吞噬功能相关[20]。抑郁症的发病机制可能与树突棘过度丢失相关,而脆性X综合征中则存在树突棘清除速率减慢的现象[21]。本研究探讨了高龄妊娠导致的子代脑功能障碍与突触修剪异常的关系,结果显示,生后第7、28、60天高龄组树突长度均明显短于适龄组,生后第28、60天树突与同心圆平均交点数亦明显少于适龄组,表明高龄妊娠可能导致子代脑发育进程中海马区的树突发育障碍,而树突复杂性降低可能促进脑内的异常神经网络形成,最终导致子代脑功能障碍。
树突棘是树突表面的棘状突起,为大部分兴奋性突触输入的接收部位。在大脑发育过程中,树突棘的密度及形态是调节突触连接强度及稳定性的决定性因素,与突触可塑性、学习及记忆机制相关,特别是在长时程增强及长时程抑制中,树突棘的结构变化与记忆形成、消失均密切相关[22-23]。目前认为,树突棘表达异常可能是神经精神疾病的病理基础,在阿尔茨海默病患者大脑中最早可观察到的结构变化是树突棘丢失[24],而在自闭症患者大脑中则发现未成熟的树突棘普遍存在[25]。本研究进一步分析高龄组出生后不同时期的树突棘密度变化,结果发现,高龄组生后第7、28、60天树突棘密度较适龄组明显降低,表明高龄妊娠子代在生后各年龄段均出现树突棘丢失。由此推测,高龄妊娠子代在大脑发育过程中存在突触修剪异常,可能由此导致树突棘被过度清除或树突棘发育障碍,但这是否与高龄妊娠导致子代脑功能障碍相关,仍有待后续实验进一步深入探究。
目前认为树突棘的形态主要分为丝状伪足型、细长型、短粗型、蘑菇型及分枝型5种。其中,丝状伪足型是成熟树突棘的前体;细长型是未成熟树突棘,更具有可塑性,主要与记忆的形成相关[26];而短粗型和蘑菇型主要介导记忆的储存及巩固,被认为是成熟树突棘,其中蘑菇型树突棘头部显著增大,主要为反复突触刺激所致,也是兴奋性突触形成的标志[27];分枝型树突棘的具体功能尚不清楚。本研究结果显示,在生后第7天,高龄妊娠子代未成熟形态树突棘密度明显高于适龄组,成熟形态树突棘密度明显低于适龄组,提示高龄妊娠可能导致子代树突棘形态发生紊乱,具体表现为未成熟型树突棘增多。但在生后第28天时,高龄组成熟形态蘑菇型树突棘密度明显高于适龄组,而生后第60天时又低于适龄组,这种一过性树突棘反应性增加后又逐渐降低的现象,提示在高龄妊娠导致海马神经元树突棘形态异常的过程中,树突棘曾试图进行代偿性的“自救”,但可能存在尚不清楚的精密调节机制导致最终挽救失败。
突触修剪受众多基因的精密调控。有研究发现,MARK1基因与树突棘的形态变化密切相关[28];树突棘形态变化可能与突触细胞骨架蛋白的表达改变有关,有研究发现,F-肌动蛋白在海马区显著上调,可能导致树突棘密度增高[29]。此外,肿瘤坏死因子[30]和脑源性神经营养因子[31]表达的变化也会影响树突棘形态的改变。本课题组前期研究发现,高龄妊娠子代出生后早期海马区脑源性神经营养因子表达降低[17],故推测脑源性神经营养因子表达降低可能是影响高龄妊娠子代树突棘形态及树突发育的机制之一。有研究发现,高龄妊娠子代海马区Arc、Egr1Fos基因表达异常,而这些基因对海马区突触连接及突触可塑性至关重要[32],因此推测高龄妊娠子代出现树突棘发育紊乱可能受多个基因调控。另有研究发现,高龄妊娠子代发育期脑内DNA甲基化修饰异常[33],可能是子代树突棘发育障碍的重要机制。寻找精准的调控机制,探索有前景的干预措施,为高龄妊娠子代脑功能障碍提供治疗靶点,是未来研究的方向。
综上所述,本研究通过建立高龄妊娠子代大鼠模型,发现高龄妊娠可能会降低子代海马树突的复杂性,使树突棘密度降低、形态紊乱,提示高龄妊娠子代在脑发育的关键时期存在突触修剪异常,这可能是导致子代脑功能障碍的重要机制,但具体分子机制尚需进一步深入研究。
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2025年第50卷第5期
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doi: 10.11855/j.issn.0577-7402.0804.2025.0218
  • 接收时间:2024-06-07
  • 首发时间:2025-10-29
  • 出版时间:2025-05-28
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  • 收稿日期:2024-06-07
  • 录用日期:2024-07-31
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National Natural Science Foundation of China(82301654)
Scientific and Technological Research Program of Chongqing Municipal Education Commission(KJQN202100423)
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    重庆医科大学附属儿童医院神经内科/国家儿童健康与疾病临床医学研究中心/儿童发育疾病研究教育部重点实验室/ 儿童神经发育与认知障碍重庆市重点实验室,重庆 400014

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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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