儿童青少年大便性状与肠道菌群多样性的关联

王兵; 吴彦; 金慧; 皮雄娥; 刘伟; 许永杰; 赵刚

doi:10.16835/j.cnki.1000-9817.2024018

儿童青少年大便性状与肠道菌群多样性的关联

doi: 10.16835/j.cnki.1000-9817.2024018

王兵¹,
吴彦¹,
金慧¹,
皮雄娥²,
刘伟²,
许永杰¹,
赵刚^1, ,

1.
浙江省杭州市疾病预防控制中心传染病防制所，310021
2.
浙江省农业科学院植物保护与微生物研究所

基金项目:

浙江省基础公益计划项目 LGN22C030008

杭州市农社发展科研重点项目 202004A20

杭州市医药卫生科技项目 A20200063

详细信息

作者简介:
王兵(1985-)，女，河南省人，博士，副主任技师，主要研究方向为肠道微生态及病原菌防控

通讯作者:
赵刚，E-mail：13588706065@163.com

利益冲突声明 所有作者声明无利益冲突。
中图分类号: R179 R442.2 Q93-34
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- 被引次数: 0
出版历程
- 收稿日期: 2023-06-19
- 修回日期: 2023-10-19
- 网络出版日期: 2024-02-01
- 刊出日期: 2024-01-25

Correlation between stool form and diversity of intestinal flora among children and adolescents

WANG Bing¹,
WU Yan¹,
JIN Hui¹,
PI Xionge²,
LIU Wei²,
XU Yongjie¹,
ZHAO Gang^{1
, ,}

1.
Infectious Disease Prevention and Control Institute, Hangzhou Center of Disease Control and Prevention, Hangzhou (310021), Zhejiang Province, China

摘要

摘要: 目的研究便秘、腹泻和健康儿童青少年肠道菌群差异和多样性变化，探讨大便性状改变与肠道菌群的相关性，为儿童青少年时期的肠道微生态研究提供科学参考。方法 2021年10月至2022年3月，采集杭州市某三甲医院收治的儿童青少年42例便秘患者、37例腹泻患者和3所普通中小学43名健康者的粪便标本，提取粪便基因组DNA进行16S rRNA基因高通量测序，对测序结果开展分析。Alpha多样性分析中，3组之间的差异比较采用Kruskal-Wallis秩和检验，两两之间比较采用FDR多重检验校正。Beta多样性分析中，3组总的差异比较采用Adonis检验方法，两两比较采用ANOSIM检验方法。LEfSe分析中，通过LDA分析(线性回归分析)获得的LDA分值。结果 Alpha多样性分析显示，腹泻组、便秘组和健康组之间的香农(Shannon)指数(4.01，3.81，4.19)和辛普森(Simpson)指数(0.05，0.06，0.04)差异均有统计学意义(H值分别为6.05，6.35，P值均<0.1)；进一步两两比较，健康组的Shannon指数和Simpson指数均高于便秘组(P值均<0.1)。Beta多样性分析显示，分组因素对组间差异的影响有统计学意义(R²=0.045，P<0.1)。群落组成分析显示，3个分组属水平上共有物种数目234个，健康组特有物种36个，腹泻组特有物种36个，便秘组特有物种48个；物种差异检验结果显示在腹泻组、便秘组和健康组之间属水平上差异有统计学意义的物种依次是厚壁菌门的毛螺菌、霍氏真杆菌、韦荣氏菌属、颤螺菌属、丁酸球菌属和葡萄球菌属(H值分别为0.000 05，0.000 16，0.000 20，0.000 21，0.000 53，0.001 39，P值均<0.1)。由KEGG丰度统计和COG功能分析可见，相同功能的基因表达丰度在不同分组间差异均无统计学意义(P值均>0.1)。结论儿童青少年大便性状不同与肠道微生物种群结构变化有关。与健康组相比，便秘或腹泻时肠道菌群平衡被打破，优势菌群发生偏移，条件致病菌丰度较高。
- 粪便 /
- 胃肠道 /
- 细菌 /
- 儿童 /
- 青少年
Abstract: Objective To investigate the differences and diversity changes in gut microbiota between children and adolescents with constipation and diarrhea, and healthy individuals, and to explore the correlation between changes in stool consistency and gut microbiota, in order to provide a scientific reference for the research on intestinal microecology among children and adolescents. Methods From October 2021 to March 2022, a total of 42 children and adolescents with constipation and 37 with diarrhea from a tertiary hospital in Hangzhou City, and 43 healthy individuals from 3 primary and secondary schools were included in this study. Fecal samples of children and adolescents were collected and then stool genomic DNA was extracted for 16S rRNA gene high-throughput sequencing, and the sequencing results were analyzed. In the analysis of alpha diversity, the Kruskal-Wallis rank sum test was used to compare the differences between the three groups, and the FDR multiple testing correction was used for pairwise comparisons. In the analysis of beta diversity, the Adonis test was used to compare the overall differences between the three groups, and the ANOSIM test was used for pairwise comparisons. In the LEfSe analysis, the LDA scores obtained through LDA analysis (linear regression analysis). Results Alpha diversity analysis showed that there were statistically significant differences in the Shannon index (4.01, 3.81, 4.19) and Simpson index (0.05, 0.06, 0.04) between the diarrhea group, constipation group, and healthy group (H=6.05, 6.35, P<0.1). Further pairwise comparison showed that the Shannon index and Simpson index of the healthy group were higher than those of the constipation group (P<0.1). Beta diversity analysis showed that the impact of grouping factors on inter-group differences was statistically significant (R²=0.045, P<0.1). Community composition analysis showed that there were 234 species in total among the three groups, and 36 unique species in the healthy group, 36 species in the diarrhea group, and 48 species in the constipation group. Species difference analysis showed significant differences in species composition at the genus level among the three groups (H=0.000 05, 0.000 16, 0.000 20, 0.000 21, 0.000 53, 0.001 39, P<0.1), including Lachnospiraceae of Firmicutes phylum, Eubacterium hallii, Veillonellaceae, Qscillospiraceae, Butyricicoccaceae and Staphylococcaceae, respectively. KEGG abundance statistics and COG functional analysis showed that there were no significant differences in gene expression abundance of the same function among the three groups (P>0.1). Conclusions The different stool consistency of children and adolescents is related to changes in gut microbiota composition. Compared to the healthy group, children with constipation or diarrhea have disrupted gut microbiota balance, with a shift in dominant bacteria and a higher abundance of opportunistic pathogens.
- Feces /
- Gastrointestinal tract /
- Bacteria /
- Child /
- Adolescent
注释:

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图 1 3组样本肠道菌群的Beta多样性分析

注：横、纵坐标表示两个选定的主坐标成分，百分比表示主坐标成分对样本组成差异的贡献值，两样本点越接近，表明两样本物种组成越相似。

Figure 1. Beta diversity analysis of gut microbiota among three groups

下载: 全尺寸图片幻灯片

图 2 3组样本肠道菌群的KEGG代谢途径分析

Figure 2. KEGG enrichment analysis of gut microbiota-associated metabolic pathways among three groups

下载: 全尺寸图片幻灯片

表 1 3组样本肠道菌群的LEfSe分析

Table 1. LEfSe analysis of gut murobiota among three groups

组别	微生物类群	LDA分值
便秘组	放线菌门	4.44
	霍氏真杆菌	4.17
	红蝽菌科	4.08
	柯林斯菌属	4.08
	Anaerostipes	4.02
腹泻组	消化链球菌	4.05
	Peptostreptococcales-Tissierellales	4.04
	罗斯氏菌	4.02
	普雷沃氏菌属	3.98
	韦荣球菌属	3.53
健康组	毛螺菌属	4.14
	Lachnoclostridium	3.52
	Butyricicoccaceae	3.51
	丁酸球菌属	3.50
	伯克氏菌目	3.37

下载: 导出CSV

表 2 3组样本肠道菌群的COG功能分类结果

Table 2. The result of COG function classification of gut microbiota among three groups

功能	健康组	便秘组	腹泻组
翻译、核糖体结构和生物发生	0.084 4	0.086 2	0.084 3
转录	0.077 8	0.079 8	0.079 4
复制、重组和修复	0.063 4	0.063 8	0.063 0
信号传导机制	0.038 4	0.038 3	0.038 4
能量生产和转换	0.059 4	0.058 8	0.059 7
碳水化合物转运和代谢	0.087 3	0.087 0	0.085 2
氨基酸转运和代谢	0.099 0	0.099 2	0.099 1
核苷酸酸转运和代谢	0.037 5	0.038 1	0.037 5
脂转运和代谢	0.023 1	0.023 3	0.023 1
无机离子转运和代谢	0.061 0	0.059 4	0.061 1
次生代谢产物的生物合成、转运和分解代谢	0.049 6	0.004 6	0.005 1
细胞运动	0.005 7	0.005 2	0.006 4
防御机制	0.027 0	0.027 3	0.026 4

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