Institute of Anatomy II, University of Jena, Teichgraben 7, 07743 Jena, Germany.
Department of Veterinary Medicine, Institute of Animal Nutrition, Freie Universität Berlin, Königin-Luise-Straße 49, 14195 Berlin, Germany.
Br J Nutr. 2014 Mar 28;111(6):1040-9. doi: 10.1017/S0007114513003498. Epub 2013 Nov 11.
Protein fermentation end products may damage the colonic mucosa, which could be counteracted by dietary inclusion of fermentable carbohydrates (fCHO). Although fermentable crude protein (fCP) and fCHO are known to affect microbial ecology, their interactive effects on epithelial barrier function are unknown. In the present study, in a 2 × 2 factorial experiment, thirty-two weaned piglets were fed low-fCP/low-fCHO (14·5 % crude protein (CP)/14·5 % total dietary fibre (TDF)), low-fCP/high-fCHO (14·8 % CP/16·6 % TDF), high-fCP/low-fCHO (19·8 % CP/14·5 % TDF) and high-fCP/high-fCHO (20·1 % CP/18·0 % TDF) diets. After 21-23 d, samples of proximal and distal colonic mucosae were investigated in Ussing chambers with respect to the paracellular and transcytotic passages of macromolecules and epithelial ion transport. The high-fCHO diets were found to reduce the permeability of the distal colon to the transcytotic marker horseradish peroxidase (HRP, 44 kDa; P <0·05) and also reduce the paracellular permeation of N-hydroxysuccinimide-biotin into the submucosa (443 Da; P <0·05), whereas that of HRP was decreased by the high-fCP diets (P <0·01). Short-circuit current (active ion transport), transepithelial resistance (barrier function) and charge selectivity were largely unaffected in both the segments. However, the high-fCP diets were found to suppress the aldosterone-induced epithelial Na channel activity (P <0·01) irrespective of fCHO inclusion. The high-fCP diets generally reduced the expression of colonic claudin-1, claudin-2 and claudin-3 (P <0·01), while that of claudin-4 was increased by the high-fCHO diets (P <0·01). The high-fCHO diets also altered the ratio between occludin forms (P <0·05) and increased the expression of tricellulin in the proximal colon, which was not observed with high-fCP diets. In conclusion, dietary fCHO and fCP exerted few and largely independent effects on functional measurements, but altered tight junction protein composition in a compensatory way, so that colonic transport and barrier properties were only marginally affected.
蛋白质发酵终产物可能会损害结肠黏膜,而膳食纤维中的可发酵碳水化合物(fCHO)可对此起到缓解作用。虽然可发酵粗蛋白(fCP)和 fCHO 会影响微生物生态,但它们对上皮屏障功能的相互影响尚不清楚。在本研究中,通过 2×2 析因实验,将 32 头断奶仔猪分别饲喂低 fCP/低 fCHO(14.5%粗蛋白(CP)/14.5%总膳食纤维(TDF))、低 fCP/高 fCHO(14.8%CP/16.6%TDF)、高 fCP/低 fCHO(19.8%CP/14.5%TDF)和高 fCP/高 fCHO(20.1%CP/18.0%TDF)日粮。21-23d 后,通过 Ussing 室法研究了近端和远端结肠黏膜的大分子跨细胞和转胞运输的细胞旁和转胞通透性。高 fCHO 日粮降低了 HRP(44kDa)的跨细胞通透性(P<0.05)和 N-羟基琥珀酰亚胺-生物素向黏膜下腔的旁细胞渗透(443Da;P<0.05),而高 fCP 日粮则降低了 HRP 的通透性(P<0.01)。短电流(主动离子转运)、跨上皮电阻(屏障功能)和电荷选择性在两个节段基本不受影响。然而,高 fCP 日粮可抑制醛固酮诱导的上皮 Na 通道活性(P<0.01),而不管 fCHO 的摄入如何。高 fCP 日粮通常会降低结肠紧密连接蛋白-1、-2 和 -3 的表达(P<0.01),而高 fCHO 日粮则会增加紧密连接蛋白-4 的表达(P<0.01)。高 fCHO 日粮还改变了紧密连接蛋白-4 形式的比例(P<0.05),并增加了近端结肠的三细胞连接蛋白的表达,而高 fCP 日粮则没有观察到这种变化。总之,膳食 fCHO 和 fCP 对功能测量的影响很小且基本独立,但以补偿的方式改变了紧密连接蛋白的组成,因此结肠的转运和屏障特性仅受到轻微影响。