孤独症谱系障碍患儿不同血清总25-羟维生素D水平与肠道菌群的差异性研究

罗欣; 庞琨; 陈建雄; 王泓哲; 徐新杰; 李兵; 贾鑫淼; 尤欣

doi:10.12290/xhyxzz.2022-0254

孤独症谱系障碍患儿不同血清总25-羟维生素D水平与肠道菌群的差异性研究

doi: 10.12290/xhyxzz.2022-0254

罗欣¹,
庞琨²,
陈建雄³,
王泓哲³,
徐新杰⁴,
李兵³,
贾鑫淼^4, ,,
尤欣^1, ,

1.
中国医学科学院北京协和医院风湿免疫科风湿免疫病学教育部重点实验室国家皮肤与免疫疾病临床医学研究中心，北京 100730
2.
温州医科大学基因组医学研究院，浙江温州 325000
3.
对外经济贸易大学信息学院，北京 100029
4.
中国医学科学院北京协和医院医学科学研究中心，北京 100730

基金项目:

中国医学科学院医学与健康科技创新工程 2017-I2M-3-017

中国医学科学院中央级公益性科研院所基本科研业务费专项资金 2019XK320030

详细信息

通讯作者:
贾鑫淼, E-mail: jiaxinmiaohappy@126.com

尤欣, E-mail: youxin@pumch.cn

中图分类号: R593;R378
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出版历程
- 收稿日期: 2022-05-13
- 录用日期: 2022-06-27
- 网络出版日期: 2022-08-02
- 刊出日期: 2022-09-30

Differences of Intestinal Flora in Children with Autism Spectrum Disorder with Different Levels of Serum Total 25-hydroxyvitamin D

LUO Xin¹,
PANG Kun²,
CHEN Jianxiong³,
WANG Hongzhe³,
XU Xinjie⁴,
LI Bing³,
JIA Xinmiao^{4
, ,},
YOU Xin^{1
, ,}

1.
Department of Rheumatology and Clinical Immunology, Key Laboratory of Rheumatology & Clinical Immunology, Ministry of Education, National Clinical Research Center for Dermatologic and Immunologic Diseases, Peking Union Medical College Hospital, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing 100730, China
2.
Institute of Genomic Medicine, Wenzhou Medical University, Wenzhou, Zhejiang 325000, China
3.
School of Information Technology & Management, University of International Business and Economics, Beijing 100029, China
4.
Medical Research Center, Peking Union Medical College Hospital, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing 100730, China

Funds:

CAMS Innovation Fund for Medical Sciences 2017-I2M-3-017

Non-profit Central Research Institute Fund of Chinese Academy of Medical Sciences 2019XK320030

More Information

Corresponding author: JIA Xinmiao, E-mail: jiaxinmiaohappy@126.com; YOU Xin, E-mail: youxin@pumch.cn

摘要

摘要: 目的探究孤独症谱系障碍(autism spectrum disorder，ASD)患儿维生素D正常与缺乏状态下的肠道菌群差异及血清总25-羟维生素D[total 25-hydroxyvitamin D，T-25(OH)D]水平与肠道菌群的相关性。方法回顾性纳入2019年10月至2022年2月于北京协和医院风湿免疫科门诊就诊的1~12岁ASD患儿的临床资料。采用液相色谱串联质谱法检测ASD患儿的血清T- 25(OH)D水平，并根据血清T- 25(OH)D水平将其分为维生素D正常组[T-25(OH)D＞30 μg/L]、不足组[20 μg/L≤T-25(OH)D ≤30 μg/L]和缺乏组[T-25(OH)D＜20 μg/L]。应用生物信息学方法分析ASD患儿的肠道宏基因组测序结果。结果共46例符合纳入和排除标准的ASD患儿纳入本研究，维生素D正常组、不足组、缺乏组分别为15例、16例、15例。线性判别分析发现，维生素D缺乏组的沃氏嗜胆菌、Adlercreutzia equolifaciens、Asaccharobacter celatus、大肠埃希菌显著升高；而脆弱拟杆菌和Hungatella hathewayi丰度显著降低。沃氏嗜胆菌和Adlercreutzia equolifaciens丰度与血清T- 25(OH)D水平均呈负相关(r=-0.45, fdr=0.055, P=0.002; r=-0.44, fdr=0.055, P=0.003)；脆弱拟杆菌丰度与血清T- 25(OH)D水平呈正相关(r=0.42, fdr=0.073, P=0.004)。结论 ASD患儿的维生素D缺乏状态可能加重肠道菌群紊乱，血清T-25(OH)水平降低可能使潜在有害菌定植增加、益生菌定植减少。本研究为ASD患儿积极补充维生素D提供了证据支持。
- 肠道微生物 /
- 孤独症谱系障碍 /
- 维生素D
Abstract: Objective To investigate the differences of intestinal flora in Vitamin D adequacy and deficiency groups of children with autism spectrum disorder(ASD) and the correlation between serum total 25-hydroxyvitamin D [T-25(OH)D] levels and intestinal flora. Methods ASD children who attended the outpatient clinic of the department of Rheumatology and Clinical Immunology of Peking Union Medical College Hospital during October 2009 and February 2022 were retrospectively included in the study. According to the serum T-25(OH)D levels, they were divided into Vitamin D adequacy group[T-25(OH)D > 30 μg/L], Vitamin D insufficiency group[20 μg/L≤T-25(OH)D≤30 μg/L] and Vitamin D deficiency group[T-25(OH)D < 20 μg/L].Serum T-25(OH)D levels were measured using liquid chromatography-tandem mass spectrometry. Human gut metagenome data from these children with ASD were analyzed using bioinformatics methods. Results 46 children with ASD who met the inclusion and exclusion criteria were included in the study. The numbers of patients in Vitamin D adequacy group, Vitamin D insufficiency group and Vitamin D deficiency group were respectively 15, 16 and 15. Linear discriminant analysis revealed that the bacteria abundance of Bilophila wadsworthia, Adlercreutzia equolifaciens, Asaccharobacter celatus and Escherichia coli were significantly enriched, while the bacteria abundance of Bacteroides fragilis and Hungatella hathewayi were significantly lower in the Vitamin D deficiency group. The relative abundance of Bilophila wadsworthia and Adlercreutzia equolifaciens were negatively correlated with serum T-25(OH)D levels(r=-0.45, fdr=0.055, P=0.002;r=-0.44, fdr=0.055, P=0.003), and the relative abundance of Bacteroides fragilis was positively correlated with serum T-25(OH)D levels (r=0.42, fdr=0.073, P=0.004). Conclusions Vitamin D deficiency in ASD may exacerbate ASD flora disorders, and decreased serum T-25(OH)D levels may facilitate potentially harmful bacteria but inhibit probiotic colonization. This study provides partial evidence that children with ASD should be actively supplemented with vitamin D.
- gut microbiota /
- autism spectrum disorder /
- vitamin D
注释:

作者贡献：罗欣负责数据处理和论文撰写；庞琨、陈建雄、王泓哲负责数据处理；徐新杰、李兵负责研究指导；贾鑫淼和尤欣负责研究设计、论文修订与审核。

利益冲突：所有作者均声明不存在利益冲突

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图 1 孤独症谱系障碍患儿肠道菌群物种丰度和香农指数箱线图

下载: 全尺寸图片幻灯片

图 2 基于加权UniFrac距离的β多样性分析

PCoA：主坐标分析

下载: 全尺寸图片幻灯片

图 3 孤独症谱系障碍患儿肠道菌群差异分类进化树

注：由内至外辐射的圆点代表了菌群由界至种的分类水平，不同水平上的每个圆点代表了该水平上的一个分类群，每个圆点的大小与该分类群的相对丰度成正比，红色圆点代表维生素D缺乏组中显著升高的分类群，绿色圆点代表维生素D正常组中显著升高的分类群，黄色圆点为无显著差异的分类群

下载: 全尺寸图片幻灯片

图 4 孤独症谱系障碍患儿肠道菌群物种水平的线性判别分析

注：LDA Score阈值为2

下载: 全尺寸图片幻灯片

图 5 血清总25-羟维生素D水平与物种相对丰度关系

下载: 全尺寸图片幻灯片

表 1 ASD患儿一般临床资料

组别	男性[n(%)]	年龄(x±s，岁)	CARS评分(x±s)		重度ASD[n(%)]
维生素D正常组(n=15)	12(80.0)	3.20±1.859	39.00±5.928		10(66.7)
维生素D不足组(n=16)	11(68.8)	3.44±1.459	41.69±6.681		12(75.0)
维生素D缺乏组(n=15)	11(73.3)	4.40±1.056	39.67±6.619		10(66.7)

组别	胃肠道症状[n(%)]	挑食[n(%)]	兴奋[n(%)]	过敏[n(%)]	情绪问题[n(%)]
维生素D正常组(n=15)	15(100)	12(80.0)	15(100)	14(93.3)	12(80.0)
维生素D不足组(n=16)	15(93.8)	15(93.8)	16(100)	12(75.0)	14(87.5)
维生素D缺乏组(n=15)	15(100)	13(86.7)	15(100)	12(80.0)	13(86.7)
ASD: 孤独症谱系障碍；CARS：儿童孤独症评定量表

下载: 导出CSV

表 2 孤独症谱系障碍患儿肠道菌群β多样性分析

分组	置换多元方差分析
分组	R²	P
维生素D缺乏组比正常组	0.0387	0.291
维生素D不足组比正常组	0.0266	0.541
维生素D不足组比缺乏组	0.0316	0.382

下载: 导出CSV

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孤独症谱系障碍患儿不同血清总25-羟维生素D水平与肠道菌群的差异性研究

doi: 10.12290/xhyxzz.2022-0254

通讯作者:
贾鑫淼, E-mail: jiaxinmiaohappy@126.com

尤欣, E-mail: youxin@pumch.cn

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Differences of Intestinal Flora in Children with Autism Spectrum Disorder with Different Levels of Serum Total 25-hydroxyvitamin D

Corresponding author: JIA Xinmiao, E-mail: jiaxinmiaohappy@126.com; YOU Xin, E-mail: youxin@pumch.cn

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孤独症谱系障碍患儿不同血清总25-羟维生素D水平与肠道菌群的差异性研究

doi: 10.12290/xhyxzz.2022-0254

通讯作者: 贾鑫淼, E-mail: jiaxinmiaohappy@126.com 尤欣, E-mail: youxin@pumch.cn

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Differences of Intestinal Flora in Children with Autism Spectrum Disorder with Different Levels of Serum Total 25-hydroxyvitamin D

Corresponding author: JIA Xinmiao, E-mail: jiaxinmiaohappy@126.com; YOU Xin, E-mail: youxin@pumch.cn

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通讯作者:
贾鑫淼, E-mail: jiaxinmiaohappy@126.com

尤欣, E-mail: youxin@pumch.cn