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Article(id=1240651442827088521, tenantId=1146029695717560320, journalId=1227665162245664772, issueId=1240651438955754377, articleNumber=null, orderNo=null, doi=10.20043/j.cnki.MPM.202404431, pmid=null, cstr=null, oa=null, hot=null, price=null, onlineType=0, articleFormat=0, articleType=null, articleTypeStr=null, receivedDate=1713888000000, receivedDateStr=2024-04-24, revisedDate=null, revisedDateStr=null, acceptedDate=null, acceptedDateStr=null, onlineDate=1773723954159, onlineDateStr=2026-03-17, pubDate=1719244800000, pubDateStr=2024-06-25, doiRegisterDate=null, doiRegisterDateStr=null, onlineIssueDate=1773723954159, onlineIssueDateStr=2026-03-17, onlineJustAcceptDate=null, onlineJustAcceptDateStr=null, onlineFirstDate=null, onlineFirstDateStr=null, sourceXml=null, magXml=null, createTime=1773723954159, creator=13701087609, updateTime=1773723954159, updator=13701087609, issue=Issue{id=1240651438955754377, tenantId=1146029695717560320, journalId=1227665162245664772, year='2024', volume='51', issue='12', pageStart='2113', pageEnd='2912', issueExtLink='null', onlineDate='null', pubDate='null', beforeIssueId=null, nextIssueId=null, price=null, status=1, issueComplete=1, articleOrder=1, issueType=-1, specialIssue=null, createTime=1773723953236, creator=13701087609, updateTime=1773723953236, updator=13701087609, preIssue=null, nextIssue=null, ext=null, issueFiles=null}, startPage=2198, endPage=2203, ext={EN=ArticleExt(id=1240651444387369653, articleId=1240651442827088521, tenantId=1146029695717560320, journalId=1227665162245664772, language=EN, title=Differential analyses of gut microbiota in infant patients with atopic dermatitis aged 0-3 years old, columnId=1228016568949474136, journalTitle=Modern Preventive Medicine, columnName=Child and Adolescent health, Maternal and Child Health, runingTitle=null, highlight=null, articleAbstract=
Objective

To investigate the differences in gut flora between infants and children aged 0~3 years with atopic dermatitis (AD) and healthy infants and children.

Methods

General information and fecal samples were collected from children with AD aged 0-3 years (AD group) and healthy infants and children (HC group). T Test and Chi-Square Tests were used to analyze the general information of infants and children in two groups, and 16S rRNA sequencing technology combined with bioinformatics analysis to compare the diversity and differences of intestinal flora in two groups.

Results

General information only history of allergy in the immediate family was significantly different between the two groups (χ2=13.875, P<0.001). The difference in the diversity of Gut microbiota between the two groups was not statistically significant. At the gate level, the relative abundance of Verrucomicrobiota was significantly lower in the AD group than in the HC group (P=0.027), whereas the relative abundance of Fusobacteriota was significantly higher than in the HC group (P=0.038). At the genus level, the relative abundance of the genera Cetobacterium (P=0.010), Dysgonomonas (P=0.013), and Fusicatenibacter (P=0.044) in the AD group was significantly higher than that in the HC group, while the relative abundance of the genera Actinomyces (P=0.029), Collinsella (P=0.017) and Eggerthella (P=0.028) were significantly lower in relative abundance than the HC group. LEfSe analysis showed that the relative abundance of Coriobacteriaceae (P=0.017) and Bifidobacterium_longum (P=0.002) in the AD group was significantly lower than that in the HC group.

Conclusion

There are some differences in the composition of the Gut microbiota between the AD and HC groups. Differential microorganisms such as Fusicatenibacter, Eggerthella, Actinomyces and Bifidobacterium_longum may have a correlation with AD development, suggesting that regulating the Gut microbiota early in life may prevent and treat AD.

, correspAuthors=Hong WANG, Guo-qing WANG, authorNote=null, correspAuthorsNote=null, copyrightStatement=null, copyrightOwner=null, 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, authorCompany=null, fund=null, authors=null, authorsList=Chen-xi LINGHU, Xian MENG, Jing CHEN, Bi-qing ZHA, Si-si DENG, Wen-jing LU, Hong WANG, Guo-qing WANG), CN=ArticleExt(id=1240651446035731243, articleId=1240651442827088521, tenantId=1146029695717560320, journalId=1227665162245664772, language=CN, title=0~3岁特异性皮炎婴幼儿患者肠道菌群差异分析, columnId=1228016570031604578, journalTitle=现代预防医学, columnName=儿少卫生与妇幼保健, runingTitle=null, highlight=null, articleAbstract=
目的

通过病例对照研究探讨0~3岁特异性皮炎（Atopic dermatitis，AD）婴幼儿与健康婴幼儿肠道菌群的差异。

方法

收集0~3岁AD患儿（AD组）与健康婴幼儿（HC组）的一般资料与粪便样品，采用统计学方法t检验和χ2检验分析两组婴幼儿的一般资料，16S rRNA测序技术联合生物信息学分析比较两组肠道菌群的多样性和差异。

结果

一般资料仅有直系亲属过敏史在两组间有显著差异（χ2=13.875，P<0.001）。两组肠道菌群多样性差异无统计学意义；在门水平上，AD组疣微菌门相对丰度显著低于HC组（P=0.027），而梭杆菌门相对丰度显著高于HC组（P=0.038）；在属水平上，AD组鲸杆菌属（P=0.010）、异常单胞菌属（P=0.013）、梭杆菌属（P=0.044）等菌属相对丰度显著高于HC组，而放线菌属（P=0.029）、柯林斯氏菌属（P=0.017）、埃格特菌属（P=0.028）等菌属相对丰度显著低于HC组。LEfSe分析结果显示AD组红蝽菌科（P=0.017）和长双歧杆菌（P=0.002）的相对丰度显著低于HC组。

结论

AD组和HC组肠道菌群组成存在一定的差异，梭杆菌属、埃格特菌属、放线菌属和长双歧杆菌等差异微生物可能与AD的发生发展具有相关性，提示在生命早期调节肠道菌群可能预防及治疗AD。

, correspAuthors=王红, 王国庆, authorNote=null, correspAuthorsNote=
王红，E-mail:；
王国庆，E-mail：
, copyrightStatement=本刊刊出的所有文章不代表中华预防医学会和本刊编委会的观点，除非特别声明。, copyrightOwner=中华预防医学会和四川大学华西公共卫生学院, extLink=null, articleAbsUrl=null, sourceXml=UDYL4T0ZxUqnSD6h0UAQcg==, magXml=d6sWfzOdiSVmQN7t8iorPA==, pdfUrl=null, pdf=JoQQqT9Rbo26DcTQbkx4Uw==, pdfFileSize=1200355, pdfExtLink=null, richHtmlUrl=null, mobilePdfUrl=null, reviewReport=null, pdfFirstPage=null, abstractGraph=/FDw2uBF5cy3KHsam4oPng==, abstractGraphContent=null, abstractVideo=null, citation=null, cebUrl=null, magXmlContent=o9LL0wYNM2X7IgqNBXZuHA==, mapNumber=null, authorCompany=null, fund=null, authors=

令狐晨曦（1998—），女，硕士在读，研究方向：卫生检验与检疫（微生物）

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令狐晨曦（1998—），女，硕士在读，研究方向：卫生检验与检疫（微生物）

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令狐晨曦（1998—），女，硕士在读，研究方向：卫生检验与检疫（微生物）

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BMC Microbiology, 2010, 10: 4., articleTitle=Impact of a synbiotic food on the gut microbial ecology and metabolic profiles, refAbstract=null), Reference(id=1240651459361034761, tenantId=1146029695717560320, journalId=1227665162245664772, articleId=1240651442827088521, doi=null, pmid=null, pmcid=null, year=2018, volume=18, issue=1, pageStart=209, pageEnd=null, url=null, language=null, rfNumber=[30], rfOrder=29, authorNames=Oki K, Akiyama T, Matsuda K, journalName=BMC Microbiology, refType=null, unstructuredReference=Oki K, Akiyama T, Matsuda K, et al. Long-term colonization exceeding six years from early infancy of Bifidobacterium longum subsp. longum in human gut[J]. BMC Microbiology, 2018, 18(1): 209., articleTitle=Long-term colonization exceeding six years from early infancy of Bifidobacterium longum subsp. longum in human gut, refAbstract=null), Reference(id=1240651459453309453, tenantId=1146029695717560320, journalId=1227665162245664772, articleId=1240651442827088521, doi=null, pmid=null, pmcid=null, year=2016, volume=11, issue=11, pageStart=null, pageEnd=null, url=null, language=null, rfNumber=[31], rfOrder=30, authorNames=Zheng HJ, Liang H, Wang YZ, journalName=PLOS One, refType=null, unstructuredReference=Zheng HJ, Liang H, Wang YZ, et al. Altered gut microbiota composition associated with eczema in infants[J]. PLOS One, 2016, 11(11): e0166026., articleTitle=Altered gut microbiota composition associated with eczema in infants, refAbstract=null), Reference(id=1240651459553972750, tenantId=1146029695717560320, journalId=1227665162245664772, articleId=1240651442827088521, doi=null, pmid=null, pmcid=null, year=2022, volume=14, issue=5, pageStart=549, pageEnd=564, url=null, language=null, rfNumber=[32], rfOrder=31, authorNames=Kim S, Han SY, Lee J, journalName=Allergy, Asthma & Immunology Research, refType=null, unstructuredReference=Kim S, Han SY, Lee J, et al. Bifidobacterium longum and Galactooligosaccharide Improve Skin Barrier Dysfunction and Atopic Dermatitis-like Skin[J]. 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注：（a）为Venn图，（b）为Specaccum物种累积曲线图

, figureFileSmall=0CETOytJlY2ksp2UEL6IaA==, figureFileBig=/FDw2uBF5cy3KHsam4oPng==, tableContent=null), ArticleFig(id=1240651453690335488, tenantId=1146029695717560320, journalId=1227665162245664772, articleId=1240651442827088521, language=EN, label=Fig.2, caption=Alpha diversity analysis, figureFileSmall=9Pw7TJGjNMhxFVW0Qf2Tdw==, figureFileBig=Bu0PTKpRx+1nR0HPmylEZw==, tableContent=null), ArticleFig(id=1240651453807776007, tenantId=1146029695717560320, journalId=1227665162245664772, articleId=1240651442827088521, language=CN, label=图2, caption=α多样性分析

注：（a）为Chao1指数，（b）为observed_species指数，（c）为Shannon指数，（d）为Simpson指数

, figureFileSmall=9Pw7TJGjNMhxFVW0Qf2Tdw==, figureFileBig=Bu0PTKpRx+1nR0HPmylEZw==, tableContent=null), ArticleFig(id=1240651454042657037, tenantId=1146029695717560320, journalId=1227665162245664772, articleId=1240651442827088521, language=EN, label=Fig.3, caption=Beta diversity analysis, figureFileSmall=JIxXP6SHn/+ZHznqdFT68g==, figureFileBig=t0BozSi9+X0ggwes5DfgBw==, tableContent=null), ArticleFig(id=1240651454243983637, tenantId=1146029695717560320, journalId=1227665162245664772, articleId=1240651442827088521, language=CN, label=图3, caption=β多样性分析, figureFileSmall=JIxXP6SHn/+ZHznqdFT68g==, figureFileBig=t0BozSi9+X0ggwes5DfgBw==, tableContent=null), ArticleFig(id=1240651454336258335, tenantId=1146029695717560320, journalId=1227665162245664772, articleId=1240651442827088521, language=EN, label=Fig.4, caption=Species composition and difference analysis of gut microbiota, figureFileSmall=nMnNNC+qtbNLztCSfDyXew==, figureFileBig=4LHA6MVMbV+c2ogb8Ie/tw==, tableContent=null), ArticleFig(id=1240651454436921639, tenantId=1146029695717560320, journalId=1227665162245664772, articleId=1240651442827088521, language=CN, label=图4, caption=肠道菌群物种组成及差异分析

注：（a）为两组门水平物种组成分布图，（b）为两组属水平物种组成分布，（c）为属水平差异物种误差图（仅显示相对丰度排名前二十的物种），（d）为差异物种LDA分布柱状图。

, figureFileSmall=nMnNNC+qtbNLztCSfDyXew==, figureFileBig=4LHA6MVMbV+c2ogb8Ie/tw==, tableContent=null), ArticleFig(id=1240651454525002027, tenantId=1146029695717560320, journalId=1227665162245664772, articleId=1240651442827088521, language=EN, label=Table 1, caption=

Basic information statistics table

, figureFileSmall=null, figureFileBig=null, tableContent=
变量AD组（n=25）HC组（n=25）t/χ2P
性别0.7640.561
男14（56）17（68）
女11（44）8（32）
年龄（岁）1.08±0.851.10±0.83650a0.808
BMI（kg/m2）17.21±1.5118.16±2.15-1.8140.076
分娩方式0.0001.000
自然分娩12（48）12（48）
剖宫产13（52）13（52）
多胞胎史1（4）1（4）0.000b1.000
宠物接触史1（4）2（8）0.000b1.000
直系亲属过敏史21（84）8（32）13.875<0.001
喂养方式1.834c0.457
母乳喂养12（48）10（40）
完全奶粉喂养0（0）2（8）
混合喂养13（52）13（52）
添加辅食时间4.5170.105
还未添加8（32）2（8）
4~6月龄7（28）9（36）
>6月龄10（40）14（56）
母乳喂养时长（月）3.216c0.37
00（0）2（8）
1~618（72）14（56）
7~125（20）8（32）
>122（8）1（4）
抗生素使用史10（36）8（28）0.3470.556
益生菌使用史20（76）19（72）0.1170.733
), ArticleFig(id=1240651454675996982, tenantId=1146029695717560320, journalId=1227665162245664772, articleId=1240651442827088521, language=CN, label=表1, caption=

基本信息统计表

, figureFileSmall=null, figureFileBig=null, tableContent=
变量AD组（n=25）HC组（n=25）t/χ2P
性别0.7640.561
男14（56）17（68）
女11（44）8（32）
年龄（岁）1.08±0.851.10±0.83650a0.808
BMI（kg/m2）17.21±1.5118.16±2.15-1.8140.076
分娩方式0.0001.000
自然分娩12（48）12（48）
剖宫产13（52）13（52）
多胞胎史1（4）1（4）0.000b1.000
宠物接触史1（4）2（8）0.000b1.000
直系亲属过敏史21（84）8（32）13.875<0.001
喂养方式1.834c0.457
母乳喂养12（48）10（40）
完全奶粉喂养0（0）2（8）
混合喂养13（52）13（52）
添加辅食时间4.5170.105
还未添加8（32）2（8）
4~6月龄7（28）9（36）
>6月龄10（40）14（56）
母乳喂养时长（月）3.216c0.37
00（0）2（8）
1~618（72）14（56）
7~125（20）8（32）
>122（8）1（4）
抗生素使用史10（36）8（28）0.3470.556
益生菌使用史20（76）19（72）0.1170.733
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0~3岁特异性皮炎婴幼儿患者肠道菌群差异分析
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令狐晨曦 1 , 孟仙 2 , 陈静 3 , 查碧晴 1 , 邓思思 4 , 卢雯静 1 , 王红 2, * , 王国庆 1, *
现代预防医学 | 儿少卫生与妇幼保健 2024,51(12): 2198-2203
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现代预防医学 | 儿少卫生与妇幼保健 2024, 51(12): 2198-2203
0~3岁特异性皮炎婴幼儿患者肠道菌群差异分析
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令狐晨曦1, 孟仙2, 陈静3, 查碧晴1, 邓思思4, 卢雯静1, 王红2, * , 王国庆1, *
作者信息
  • 1.四川大学华西公共卫生学院/华西第四医院，四川 成都 610041
  • 2.四川省妇幼保健院
  • 3.成都东部新区公共卫生中心
  • 4.湖南省疾病预防控制中心

    • 令狐晨曦（1998—），女，硕士在读，研究方向：卫生检验与检疫（微生物）

通讯作者:

王红，E-mail:；
王国庆，E-mail：
Differential analyses of gut microbiota in infant patients with atopic dermatitis aged 0-3 years old
Chen-xi LINGHU1, Xian MENG2, Jing CHEN3, Bi-qing ZHA1, Si-si DENG4, Wen-jing LU1, Hong WANG2, * , Guo-qing WANG1, *
Affiliations
  • West China School of Public Health and West China Fourth Hospital, Sichuan University, Chengdu, Sichuan 610041, China
出版时间: 2024-06-25 doi: 10.20043/j.cnki.MPM.202404431
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摘要
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目的

通过病例对照研究探讨0~3岁特异性皮炎（Atopic dermatitis，AD）婴幼儿与健康婴幼儿肠道菌群的差异。

方法

收集0~3岁AD患儿（AD组）与健康婴幼儿（HC组）的一般资料与粪便样品，采用统计学方法t检验和χ2检验分析两组婴幼儿的一般资料，16S rRNA测序技术联合生物信息学分析比较两组肠道菌群的多样性和差异。

结果

一般资料仅有直系亲属过敏史在两组间有显著差异（χ2=13.875，P<0.001）。两组肠道菌群多样性差异无统计学意义；在门水平上，AD组疣微菌门相对丰度显著低于HC组（P=0.027），而梭杆菌门相对丰度显著高于HC组（P=0.038）；在属水平上，AD组鲸杆菌属（P=0.010）、异常单胞菌属（P=0.013）、梭杆菌属（P=0.044）等菌属相对丰度显著高于HC组，而放线菌属（P=0.029）、柯林斯氏菌属（P=0.017）、埃格特菌属（P=0.028）等菌属相对丰度显著低于HC组。LEfSe分析结果显示AD组红蝽菌科（P=0.017）和长双歧杆菌（P=0.002）的相对丰度显著低于HC组。

结论

AD组和HC组肠道菌群组成存在一定的差异，梭杆菌属、埃格特菌属、放线菌属和长双歧杆菌等差异微生物可能与AD的发生发展具有相关性，提示在生命早期调节肠道菌群可能预防及治疗AD。

特异性皮炎  /  肠道菌群  /  16S rRNA测序
Objective

To investigate the differences in gut flora between infants and children aged 0~3 years with atopic dermatitis (AD) and healthy infants and children.

Methods

General information and fecal samples were collected from children with AD aged 0-3 years (AD group) and healthy infants and children (HC group). T Test and Chi-Square Tests were used to analyze the general information of infants and children in two groups, and 16S rRNA sequencing technology combined with bioinformatics analysis to compare the diversity and differences of intestinal flora in two groups.

Results

General information only history of allergy in the immediate family was significantly different between the two groups (χ2=13.875, P<0.001). The difference in the diversity of Gut microbiota between the two groups was not statistically significant. At the gate level, the relative abundance of Verrucomicrobiota was significantly lower in the AD group than in the HC group (P=0.027), whereas the relative abundance of Fusobacteriota was significantly higher than in the HC group (P=0.038). At the genus level, the relative abundance of the genera Cetobacterium (P=0.010), Dysgonomonas (P=0.013), and Fusicatenibacter (P=0.044) in the AD group was significantly higher than that in the HC group, while the relative abundance of the genera Actinomyces (P=0.029), Collinsella (P=0.017) and Eggerthella (P=0.028) were significantly lower in relative abundance than the HC group. LEfSe analysis showed that the relative abundance of Coriobacteriaceae (P=0.017) and Bifidobacterium_longum (P=0.002) in the AD group was significantly lower than that in the HC group.

Conclusion

There are some differences in the composition of the Gut microbiota between the AD and HC groups. Differential microorganisms such as Fusicatenibacter, Eggerthella, Actinomyces and Bifidobacterium_longum may have a correlation with AD development, suggesting that regulating the Gut microbiota early in life may prevent and treat AD.

Atopic dermatitis  /  Gut microbiota  /  16S rRNA sequencing
令狐晨曦, 孟仙, 陈静, 查碧晴, 邓思思, 卢雯静, 王红, 王国庆. 0~3岁特异性皮炎婴幼儿患者肠道菌群差异分析. 现代预防医学, 2024 , 51 (12) : 2198 -2203 . DOI: 10.20043/j.cnki.MPM.202404431
Chen-xi LINGHU, Xian MENG, Jing CHEN, Bi-qing ZHA, Si-si DENG, Wen-jing LU, Hong WANG, Guo-qing WANG. Differential analyses of gut microbiota in infant patients with atopic dermatitis aged 0-3 years old[J]. Modern Preventive Medicine, 2024 , 51 (12) : 2198 -2203 . DOI: 10.20043/j.cnki.MPM.202404431
正文
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特异性皮炎（Atopic dermatitis，AD）是一种瘙痒性炎症性皮肤病，病程缓慢易复发[1]。研究显示，AD在儿童中的患病率高达15%~20%，在成人中患病率为10% [2]。AD患者会因瘙痒、疼痛、睡眠困难等症状影响其生活质量，同时AD的持续性治疗也会给患者带来较大的经济负担[3]。AD的发病机制尚不明确，可能归因于环境暴露、遗传因素、免疫功能障碍、皮肤屏障受损以及微生物失衡等因素[4]。肠道在免疫反应中起着重要作用，免疫细胞和肠道菌群相互调节以维持肠道稳态和正常免疫功能[5]。有研究揭示了肠道菌群与AD的相关性，推测肠道紊乱可能与AD的发生和严重程度相关，同时有研究表明过敏性疾病患儿和健康婴幼儿肠道菌群的组成存在显著差异[6-7]。随着“肠道-皮肤”轴理论的提出，靶向肠道菌群改变被认为可能是调节免疫反应和改善皮肤炎症的新策略[8]。目前关于婴幼儿肠道菌群与AD的研究相对较少，据研究报道，婴幼儿在1岁时具有其各自的肠道微生物群，到3岁时逐渐向成人的肠道微生物群转变[9-10]。因此本研究通过病例对照研究分析0~3岁AD患儿与健康婴幼儿肠道菌群的组成差异，探究肠道微生物与AD发生发展的相关性，为AD的防治提供理论依据。
1　材料与方法
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1.1　研究对象
选取2021年11月—2022年4月在四川省妇幼保健院皮肤科门诊就诊的0~3岁特异性皮炎患者作为病例组（AD组），同时选取在该院儿童保健科门诊进行健康体格检查的婴幼儿作为健康对照组（HC组）。本研究经过四川大学华西公共卫生学院医学伦理委员会的审批（批件号：GWll2021061），研究对象的家长或监护人自愿参加并签署知情同意书。
1.2　纳入标准和排除标准
1.2.1　纳入标准
AD组：（1）年龄为0~3岁；（2）经专业医生诊断为AD患者并符合Williams诊断标准[11]，包括皮肤瘙痒与干燥、典型屈侧受累史（膝窝、肘窝、围绕颈周或踝前）以及个人或家族中有过敏性疾病史等；(3)无其他疾病，包括皮肤疾病、器官或系统严重异常等疾病。
HC组：(1)无过敏性疾病，包括AD、过敏性哮喘、食物过敏等；(2)年龄与其他疾病情况同AD组。
1.2.2　排除标准
AD组：(1)患有免疫系统或免疫缺陷相关疾病；(2)患有炎症性肠病、胃肠炎及肠易激综合征疾病，或自出生以来做过胃肠道大手术；(3)近两周内患有腹泻等胃肠道疾病，使用过含碱式水杨酸铋或类似成分的药物；(4)近两周内通过各种途径摄入过抗生素；(5)近两周内频繁（每周三次及以上）服用益生菌或益生元等制剂。
HC组：同AD组。
1.3　方法
1.3.1　一般的资料收集
向研究对象的监护人发放问卷，通过问卷调查的方式收集研究对象的一般资料信息。问卷内容包括研究对象的年龄、性别、直系亲属过敏史、出生方式、喂养方式、抗生素及益生菌的使用情况等。
1.3.2　粪便样品采集
粪便样品由研究对象的监护人严格按照采样说明要求，使用研究人员提供的采样用具进行采样，其中采样用具包括无菌采样管、洁净塑封袋、冰袋、保温袋、采样说明。粪便样品采集后在2 h内尽快运送回实验室，分装后于-80℃保存备用。
1.3.3　肠道菌群分析
采集粪便样品后由奥维森基因科技有限公司进行DNA提取、PCR扩增及测序，扩增测序区域为细菌16S rRNA V3～V4可变区。测序数据经过质控、过滤、拼接和去嵌合体得到优质序列，再进行数据的OTUs（Operational Taxonomic Units）聚类和注释分析。基于聚类结果进行α和β多样性分析，基于注释结果得到各个水平的物种信息，包括物种组成和相对丰度。采用Tukey检验分析两组间α多样性指数的差异，采用Wilcoxon检验分析两组间的物种差异，检验水准为α=0.05。
1.4　统计学分析
数据分析使用SPSS 22.0软件进行，计量资料使用表示，符合正态分布及方差齐性的资料采用t检验进行分析，反之则使用Wilcoxon检验。计数资料用率或构成比表示，采用χ2检验分析，检验水准为α=0.05。
2　结果
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2.1　研究对象的一般资料
根据粪便样品收集的实际情况，研究共纳入AD患者25人，HC组25人，共计50个粪便样本进行测序分析。表1为测序粪便样品婴幼儿的特性统计，结果显示，仅有直系亲属过敏史在两组间的差异具有统计学意义，为AD组显著高于HC组（χ2=13.875，P<0.001）。
2.2　测序样本基本信息
本研究通过对25个AD患者和25个健康对照者的粪便标本进行测序检测后，共获得996 000条优质序列。对这些优质序列进行OTUs聚类分析，按97%的相似水平进行归类操作共得到3 198个OTUs。根据OTUs聚类分析结果绘制Venn图，可得到各组样本中所共有和独有的OTU数目。如图1（a）所示，其中两组共有2 061个OTUs，AD组特有454个OTUs，HC组特有683个OTUs。
2.3　Specaccum物种累积曲线
物种累积曲线可用于判断测序样本量是否充足及估计物种的丰富度。如图1（b）所示，Specaccum物种累积曲线趋于平缓，表明样本量充足，物种丰富度趋于饱和。
2.4　AD组与HC组肠道菌群分析
2.4.1　肠道菌群α多样性分析
α多样性分析可用于估计样本群落的物种丰富度和多样性。本研究选取Chao1指数、observed_species指数、Shannon指数和Simpson指数进行分析。如图2所示，四种指数均显示出AD组低于HC组的趋势，差异虽无显著性，但能在一定程度上提示AD儿童肠道菌群的丰富度和多样性低于健康儿童。
2.4.2　肠道菌群β多样性分析
PCoA分析（principal co-ordinates analysis）是基于不同距离研究样本间微生物群落组成的差异性或相似性。如图3所示，在基于bray-curtis距离的PCoA分析中，AD组和HC组菌群结构有较大部分重叠，经Permanova进一步分析结果显示其差异并无统计学意义（P>0.05）。
2.4.3　肠道菌群相对丰度差异分析
2.4.3.1　门水平肠道菌群相对丰度差异分析
图4（a）表示在门水平上两组物种相对丰度排名前20的优势菌门，主要包括厚壁菌门（Firmicutes）、放线菌门（Actinobacteriota）、拟杆菌门（Bacteroidota）、变形菌门（Proteobacteria）、疣微菌门（Verrucomicrobiota）、蓝藻菌门（Cyanobacteria）、梭杆菌门（Fusobacteriota）等。AD组中厚壁菌门、拟杆菌门、变形菌门、蓝藻菌门、梭杆菌门相对丰度高于HC组，而放线菌门、疣微菌门水平低于HC组，Wilcoxon检验结果显示仅有疣微菌门与梭杆菌门的差异具有统计学意义（P<0.05）。
2.4.3.2　属水平肠道菌群相对丰度差异分析
如图4（b）所示，在属水平上，排名前10的优势菌属中AD组拟杆菌属（Bacteroides）、布劳特氏菌属（Blautia）、埃希氏-志贺菌属（Escherichia-Shigella）、链球菌属（Streptococcus）、Clostridium_sensu_stricto_1、副拟杆菌属（Parabacteroides）相对丰度水平高于HC组，而双歧杆菌属（Bifidobacterium）、阿克曼菌属（Akkermansia）、罗氏菌属（Roseburia）、韦荣氏球菌属（Veillonella）相对丰度水平低于HC组，其差异均无显著性（P>0.05）。因此进一步通过Wilcoxon检验来分析两组属水平上的差异性物种，如图4（c）展示了相对丰度排名前20的差异菌属，AD组中鲸杆菌属（Cetobacterium）、异常单胞菌属（Dysgonomonas）、梭杆菌属（Fusicatenibacter）等菌属相对丰度显著高于HC组，而放线菌属（Actinomyces）、柯林斯氏菌（Collinsella）、埃格特菌属（Eggerthella）等菌属相对丰度显著低于HC组，其差异均具有统计学意义（P<0.05）。
2.4.3.3　LEfSe分析
LEfSe (Linear discriminant analysis Effect Size)分析用于比较两组间及组内在各水平上丰度具有显著差异的物种。本研究线性判别分析（LDA）阈值设定为3，分析结果如图4（d）所示，异常单胞菌属、梭杆菌科（Fusobacteriaceae）、梭杆菌门等在AD组中富集，长双歧杆菌（Bifidobacterium_longum）、疣微菌门、红蝽菌科（Coriobacteriaceae）、柯林斯氏菌属（Collinsella）等在HC组中富集。
3　讨论
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由于绝大部分AD病例都始发于生命早期，甚至持续到成年仍然继续发病，因此在生命早期探讨AD患者与健康人群的肠道菌群差异显得尤为重要。本研究通过分析0~3岁健康婴幼儿与AD患儿肠道菌群的差异，为临床通过调节肠道菌群防治AD提供依据。本研究一般资料分析结果显示家族过敏史具有统计学差异，与既往的研究结果一致，提示遗传因素与AD的发生存在相关性[12]。可能包括皮肤结构蛋白丝聚蛋白（Filaggrin，FLG）基因功能突变或特异性家族史两个遗传危险因素[13]。测序信息显示AD组OTUs少于HC组，结合肠道菌群α多样性分析结果，提示AD患儿的肠道菌群多样性低于健康婴幼儿，这与Penders等人[14]的研究一致。造成本研究多样性分析差异不具有统计学意义的原因可能是样本量偏少，其次小于3岁的婴幼儿的肠道微生物群处于动态发展中，分娩方式、喂养类型和生活方式等外部因素也存在一定的影响[15]。因此在样本量充足的条件下，可以根据年龄或不同的生活方式进行分层分析。
本研究通过比较AD组和HC组肠道菌群各分类水平物种组成差异发现，在门水平上的优势菌群为厚壁菌门、拟杆菌门和放线菌门等，这与Yu等人[16]的研究一致，其中疣微菌门和梭杆菌门在AD组与HC组间的差异具有显著性。阿克曼菌属是疣微菌门的代表菌属，已有研究发现阿克曼菌属可通过调节Th1和Th2细胞因子之间的平衡并调节皮肤稳态和肠道屏障功能来改善AD小鼠的病变，因此阿克曼菌属可能是治疗AD的潜在微生物靶点[17]。本研究发现梭杆菌门中的梭杆菌属和鲸杆菌属在AD组中显著富集。Wang等人[18]研究发现AD患者口服混合益生菌后梭杆菌属相对丰度显著降低。且梭杆菌属已被证实与牙周病[19]、食管癌[20]和结肠癌[21]等疾病密切相关，其致病机制可能与p38 MAPK信号通路引起炎性细胞因子增加有关[22]。鲸杆菌属是一种乙酸盐生产者，在鱼体内可通过副交感神经激活介导调节葡萄糖稳态[23]。本研究发现在AD患儿中鲸杆菌属显著增加，但由于目前鲸杆菌属在人类相关疾病方面的研究较少，因此具体的作用机制需进一步探讨。本研究结果显示AD组拟杆菌门下的异常单胞菌属显著高于HC组。Nakajima等人[24]发现异常单胞菌属与薄纤维帽粥样硬化症相关，与炎症因子IL-6水平呈显著正相关。但目前关于异常单胞菌属与AD的研究较少，其与AD发生发展是否存在相关性还需进一步研究确认。
经差异检验和LEfSe分析结果发现，放线菌门下多种物种在HC组中富集，主要包括红蝽菌科、放线菌属、柯林斯氏菌属、埃格特菌属、长双歧杆菌。Jin等人[25]研究发现红蝽菌科对过敏性鼻炎具有保护作用。柯林斯氏菌属和埃格特菌属均属于红蝽菌科，Zhou等人[26]研究发现柯林斯氏菌属在过敏性鼻炎患者中的丰度显著低于健康对照组；Reddel等人[27]研究发现埃格特菌属和放线菌属在AD患儿中的丰度低于健康儿童，与本研究结果一致。但目前关于红蝽菌科、柯林斯氏菌属和埃格特菌属功能的研究相对较少，因此它们与过敏性疾病的关系还需进一步探讨。双歧杆菌属在出生最初几天和几周内就可通过母乳喂养定植于肠道内，在人体内具有维生素产生、抑制潜在病原菌、免疫系统刺激等作用[28-29]。其中长双歧杆菌被认为是婴儿肠道中含量最丰富的物种，并可长期定植于肠道内[30]。Zheng等人[31]研究显示，与湿疹婴儿相比，长双歧杆菌在健康婴儿中富集，与本研究结果一致。研究发现，长双歧杆菌可通过抑制Th2型细胞因子的表达并降低IgE水平来调节免疫反应，并可通过口服长双歧杆菌改善AD小鼠的皮肤屏障功能和AD样皮肤[32]。因此这些差异微生物均可能是治疗AD的潜在微生物靶点。
综上所述，0~3岁AD患儿与健康婴幼儿的肠道菌群组成存在一定的差异，梭杆菌属、异常单胞菌属、柯林斯氏菌属、埃格特菌属、放线菌属和长双歧杆菌等肠道差异微生物可能与AD的发生发展相关，因此在生命早期及时调节肠道菌群可能是一种很有前途的预防及治疗AD的方法。但还需进一步关注肠道差异微生物调节AD炎症的具体机制，并结合肠道菌群代谢产物进行分析。
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  • 四川省科技厅重点研发项目(2019YFS0304)
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2024年第51卷第12期
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doi: 10.20043/j.cnki.MPM.202404431
  • 接收时间：2024-04-24
  • 首发时间：2026-03-17
  • 出版时间：2024-06-25
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  • 收稿日期：2024-04-24
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四川省科技厅重点研发项目(2019YFS0304)
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    1.四川大学华西公共卫生学院/华西第四医院，四川 成都 610041
    2.四川省妇幼保健院
    3.成都东部新区公共卫生中心
    4.湖南省疾病预防控制中心

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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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