Duodenal and faecal microbiota of celiac children: molecular, phenotype and metabolome characterization.
Duodenal and faecal microbiota of celiac children: molecular, phenotype and metabolome characterization.
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DOI：
10.1186/1471-2180-11-219
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发表时间：
2011-10-04
期刊：
影响因子：
4.2
通讯作者：
Francavilla R
中科院分区：
文献类型：
--
作者：
Di Cagno R;De Angelis M;De Pasquale I;Ndagijimana M;Vernocchi P;Ricciuti P;Gagliardi F;Laghi L;Crecchio C;Guerzoni ME;Gobbetti M;Francavilla R
乳糜泻（CD）的流行病学正在增加。乳糜泻主要表现为儿童早期小肠绒毛萎缩和吸收不良。与健康个体相比，乳糜泻患者的特点似乎是革兰氏阴性菌数量较多，革兰氏阳性菌数量较少。本研究旨在调查19名乳糜泻儿童（治疗乳糜泻，T-CD）和15名非乳糜泻儿童（HC）的微生物群和代谢组。采用通用引物和群体特异性引物对十二指肠活检和粪便样本进行pcr -变性梯度凝胶电泳（DGGE）分析。从PCR-DGGE条带数来看，T-CD患儿十二指肠活检中真细菌的多样性高于HC患儿。双歧杆菌仅在粪便样本中发现。除了少数例外，粪便样品的PCR-DGGE图谱在T-CD和HC之间存在差异。培养依赖法结果显示，HC患儿粪便样品中乳酸菌、肠球菌和双歧杆菌的水平明显高于T-CD患儿（P = 0.028、P = 0.019和P = 0.023）。与HC相比，T-CD患儿推测的拟杆菌、葡萄球菌、沙门氏菌、志贺氏菌和克雷伯氏菌的细胞计数（CFU/ml）显著升高（P = 0.014）。粪肠球菌和植物乳杆菌是最广泛鉴定的物种。在T-CD和HC儿童的所有十二指肠活检中也发现了后者。其他细菌种类仅在T-CD或HC粪便样本中被鉴定出来。随机扩增多态性DNA-PCR分析结果显示，T-CD组（26.5%）与HC组（34.6%）鉴定出乳酸菌的比例差异显著（P = 0.011）。采用气相色谱-质谱-固相微萃取和核磁共振技术对T-CD和HC患儿的粪便和尿液进行代谢组学研究。根据主坐标的典型判别分析，粪便和/或尿液样本中挥发性有机化合物和游离氨基酸的水平明显受到乳糜泻的影响。根据平行微生物学和代谢组方法显示，持续至少两年的无麸质饮食并没有完全恢复乳糜泻儿童的微生物群，因此，代谢组也没有完全恢复。一些分子（如乙酸乙酯和乙酸辛酯，一些短链脂肪酸和游离氨基酸，以及谷氨酰胺）似乎是CD的代谢特征。
Epidemiology of celiac disease (CD) is increasing. CD mainly presents in early childhood with small intestinal villous atrophy and signs of malabsorption. Compared to healthy individuals, CD patients seemed to be characterized by higher numbers of Gram-negative bacteria and lower numbers Gram-positive bacteria. This study aimed at investigating the microbiota and metabolome of 19 celiac disease children under gluten-free diet (treated celiac disease, T-CD) and 15 non-celiac children (HC). PCR-denaturing gradient gel electrophoresis (DGGE) analyses by universal and group-specific primers were carried out in duodenal biopsies and faecal samples. Based on the number of PCR-DGGE bands, the diversity of Eubacteria was the higher in duodenal biopsies of T-CD than HC children. Bifidobacteria were only found in faecal samples. With a few exceptions, PCR-DGGE profiles of faecal samples for Lactobacillus and Bifidobacteria differed between T-CD and HC. As shown by culture-dependent methods, the levels of Lactobacillus, Enterococcus and Bifidobacteria were confirmed to be significantly higher (P = 0.028; P = 0.019; and P = 0.023, respectively) in fecal samples of HC than in T-CD children. On the contrary, cell counts (CFU/ml) of presumptive Bacteroides, Staphylococcus, Salmonella, Shighella and Klebsiella were significantly higher (P = 0.014) in T-CD compared to HC children. Enterococcus faecium and Lactobacillus plantarum were the species most diffusely identified. This latter species was also found in all duodenal biopsies of T-CD and HC children. Other bacterial species were identified only in T-CD or HC faecal samples. As shown by Randomly Amplified Polymorphic DNA-PCR analysis, the percentage of strains identified as lactobacilli significantly (P = 0.011) differed between T-CD (ca. 26.5%) and HC (ca. 34.6%) groups. The metabolome of T-CD and HC children was studied using faecal and urine samples which were analyzed by gas-chromatography mass spectrometry-solid-phase microextraction and 1H-Nuclear Magnetic Resonance. As shown by Canonical Discriminant Analysis of Principal Coordinates, the levels of volatile organic compounds and free amino acids in faecal and/or urine samples were markedly affected by CD. As shown by the parallel microbiology and metabolome approach, the gluten-free diet lasting at least two years did not completely restore the microbiota and, consequently, the metabolome of CD children. Some molecules (e.g., ethyl-acetate and octyl-acetate, some short chain fatty acids and free amino acids, and glutamine) seems to be metabolic signatures of CD.
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影响因子：
12.6
作者：
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通讯作者：
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