Yeruva Laxmi, Spencer Nicole E, Saraf Manish K, Hennings Leah, Bowlin Anne K, Cleves Mario A, Mercer Kelly, Chintapalli Sree V, Shankar Kartik, Rank Roger G, Badger Thomas M, Ronis Martin J J
Arkansas Children's Nutrition Center, 15 Children's Way, Little Rock, AR, 72202, USA.
Arkansas Children's Hospital Research Institute, Little Rock, USA.
BMC Gastroenterol. 2016 Mar 22;16:40. doi: 10.1186/s12876-016-0456-x.
Breastfeeding is associated with a variety of positive health outcomes in children and is recommended exclusively for the first 6 months of life; however, 50-70 % of infants in the US are formula-fed. To test the hypothesis that immune system development and function in neonates and infants are significantly influenced by diet, 2-day old piglets were fed soy or milk formula (n = 6/group/gender) until day 21 and compared to a sow-fed group (n = 6/gender).
Histomorphometric analyses of ileum, jejunum and Peyer's patches were carried out, to determine the inflammation status, mRNA and protein expression of pro-inflammatory, anti-inflammatory and growth-related chemokines and cytokines.
In formula-fed animals, increases in ileum and jejunum villus height and crypt depth were observed in comparison to sow-fed animals (jejunum, p < 0.01 villus height, p < 0.04 crypt depth; ileum p < 0.001 villus height, p < 0.002 crypt depth). In formula-fed the lymphoid follicle size (p < 0.01) and germinal centers (p < 0.01) with in the Peyer's patch were significantly decreased in comparison to sow-fed, indicating less immune education. In ileum, formula diet induced significant up-regulation of AMCFII, IL-8, IL-15, VEGFA, LIF, FASL, CXCL11, CCL4, CCL25 and down-regulation of IL-6, IL-9, IL-10, IL-27, IFNA4, CSF3, LOC100152038, and LOC100736831 at the transcript level. We have confirmed some of the mRNA data by measuring protein, and significant down-regulation of anti-inflammatory molecule IL-10 in comparison to sow-fed piglets was observed. To further determine the membrane protein expression in the ileum, VE-cadherin, occludin, and claudin-3, Western blot analyses were conducted. Sow fed piglets showed significantly more VE-Cadherin, which associated with levels of calcium, and putrescine measured. It is possible that differences in GI tract and immune development are related to shifts in the microbiome; notably, there were 5-fold higher amounts of Lactobacillaceae spp and 3 fold higher Clostridia spp in the sow fed group in comparison to milk formula-fed piglets, whereas in milk formula-fed pigs Enterobacteriaceae spp was 5-fold higher.
In conclusion, formula diet alters GI morphology, microbial abundance, intestinal barrier protein VE-cadherin and anti-inflammatory molecule IL-10 expression. Further characterization of formula effects could lead to modification of infant formula to improve immune function, reduce inflammation and prevent conditions such as allergies and infections.
母乳喂养与儿童多种积极的健康结果相关，且建议在婴儿出生后的前6个月进行纯母乳喂养；然而，美国50%-70%的婴儿是用配方奶粉喂养的。为了验证新生儿和婴儿的免疫系统发育及功能受饮食显著影响这一假设，给2日龄仔猪分别喂食大豆配方奶粉或牛奶配方奶粉（每组/每种性别n = 6），持续至第21天，并与母猪喂养组（每种性别n = 6）进行比较。
对回肠、空肠和派伊尔结进行组织形态计量学分析，以确定炎症状态、促炎、抗炎和生长相关趋化因子及细胞因子的mRNA和蛋白质表达。
与母猪喂养的动物相比，配方奶粉喂养的动物回肠和空肠绒毛高度增加，隐窝深度增加（空肠，绒毛高度p < 0.01，隐窝深度p < 0.04；回肠，绒毛高度p < 0.001，隐窝深度p < 0.002）。与母猪喂养的相比，配方奶粉喂养的派伊尔结内淋巴滤泡大小（p < 0.01）和生发中心（p < 0.01）显著减小，表明免疫教育较少。在回肠中，配方奶粉饮食在转录水平上显著上调了AMCFII、IL-8、IL-15、VEGFA、LIF、FASL、CXCL11、CCL4、CCL25，并下调了IL-6、IL-9、IL-10、IL-27、IFNA4、CSF3、LOC100152038和LOC100736831。我们通过测量蛋白质证实了部分mRNA数据，并且观察到与母猪喂养的仔猪相比，抗炎分子IL-10显著下调。为了进一步确定回肠中的膜蛋白表达，进行了VE-钙黏蛋白、闭合蛋白和紧密连接蛋白-3的蛋白质印迹分析。母猪喂养的仔猪显示出显著更多的VE-钙黏蛋白，这与所测量的钙和腐胺水平相关。胃肠道和免疫发育方面的差异可能与微生物群的变化有关；值得注意的是，与牛奶配方奶粉喂养的仔猪相比，母猪喂养组中乳酸杆菌属的数量高5倍，梭菌属的数量高3倍，而在牛奶配方奶粉喂养的仔猪中肠杆菌科的数量高5倍。
总之，配方奶粉饮食改变了胃肠道形态、微生物丰度、肠道屏障蛋白VE-钙黏蛋白和抗炎分子IL-10的表达。对配方奶粉影响的进一步表征可能会促使婴儿配方奶粉得到改进，以提高免疫功能、减轻炎症并预防过敏和感染等病症。
