Department of Surgery, The University of Chicago Medical Center, Chicago, IL 60637, USA.
Ann Surg. 2013 Feb;257(2):287-94. doi: 10.1097/SLA.0b013e318262a6a6.
Here, we determine how formula feeding impacts the gut microbiota and host transcriptome.
Formula-fed (FF) infants are at risk for diseases that involve complex interactions between microbes and host immune elements such as necrotizing enterocolitis. The aims of this study were to simultaneously examine the microbiota and host transcriptional profiles of FF and maternal-fed (MF) mice to evaluate how diet impacts gut colonization and host genes.
After 72 hours of FF or MF, colonic tissue was collected. 16S ribosomal RNA was sequenced with Roche GS-FLX (Genome Sequencer-FLX) pyrosequencing. Operational taxonomical unit clustering, diversity analysis, and principal coordinate analysis (PCA) were performed. Complementary DNA libraries were sequenced by Solexa. Reads were annotated by BLAST (Basic Local Alignment Search Tool) search against mouse RNA database [National Center for Biotechnology Information (NCBI) build-37] and functionally classified using the KOG (Eukaryotic Orthologous Groups) database (NCBI).
Firmicutes (P < 0.001) was the dominant phylum in MF pups, whereas Proteobacteria (P < 0.001) and Bacteroidetes (P < 0.05) were dominant in FF mice. On the genus level, FF mice had increased Serratia (P < 0.001) and Lactococcus (P < 0.05) whereas MF mice had increased Lactobacillus (P < 0.001). PCA confirmed clustering by diet. Solexa sequencing demonstrated different (P < 0.05) messenger RNA transcript levels in 148 genes. Heme oxygenase 1 (P < 0.01), an oxidative stress marker, was increased 25-fold in FF mice. In addition, decreased vinculin (P < 0.05), a cytoskeletal protein associated with adherens junctions in FF pups suggested impaired gut structural integrity. Diet also impacted immune regulation, cell cycle control/gene expression, cell motility, and vascular function genes.
FF shifted gut microbiota and structural integrity, oxidative stress, and immune function genes, presumably increasing vulnerability to disease in FF mice. Interrogation of microbial and host gene expression in FF neonates may offer new insight on how diet affects disease pathogenesis.
本研究旨在探讨配方奶喂养对肠道微生物群和宿主转录组的影响。
配方奶喂养（FF）的婴儿易患与微生物和宿主免疫成分之间复杂相互作用相关的疾病，如坏死性小肠结肠炎。本研究的目的是同时检测 FF 和母乳喂养（MF）小鼠的微生物群和宿主转录谱，以评估饮食如何影响肠道定植和宿主基因。
在 FF 或 MF 喂养 72 小时后，收集结肠组织。使用罗氏 GS-FLX（基因组测序器-FLX）焦磷酸测序对 16S 核糖体 RNA 进行测序。进行操作分类单元聚类、多样性分析和主坐标分析（PCA）。互补 DNA 文库通过 Solexa 测序。通过与小鼠 RNA 数据库（美国国立生物技术信息中心[NCBI]构建 37）的 BLAST（基本局部比对搜索工具）搜索对读取进行注释，并使用 KOG（真核同源群）数据库（NCBI）对其进行功能分类。
厚壁菌门（Firmicutes）（P < 0.001）是 MF 幼崽的优势门，而变形菌门（Proteobacteria）（P < 0.001）和拟杆菌门（Bacteroidetes）（P < 0.05）是 FF 小鼠的优势门。在属水平上，FF 小鼠中 Serratia（P < 0.001）和 Lactococcus（P < 0.05）增加，而 MF 小鼠中 Lactobacillus（P < 0.001）增加。PCA 证实了饮食聚类。Solexa 测序显示 148 个基因的信使 RNA 转录水平不同（P < 0.05）。血红素加氧酶 1（Heme oxygenase 1）（P < 0.01），一种氧化应激标志物，在 FF 小鼠中增加了 25 倍。此外，FF 幼崽中粘连蛋白（Vinculin）（P < 0.05）减少，这是一种与黏附连接相关的细胞骨架蛋白，提示肠道结构完整性受损。饮食还影响免疫调节、细胞周期控制/基因表达、细胞迁移和血管功能基因。
FF 改变了肠道微生物群和结构完整性、氧化应激和免疫功能基因，可能增加了 FF 小鼠对疾病的易感性。对 FF 新生儿的微生物和宿主基因表达的研究可能为饮食如何影响疾病发病机制提供新的见解。
