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Influence of Nitrogen Levels on Nutrient Transporters and Regulators of Protein Synthesis in Small Intestinal Enterocytes of Piglets

Tian, Zhimei, Ma, Xianyong, Deng, Dun, Cui, Yiyan, Chen, Weidong
Journal of agricultural and food chemistry 2019 v.67 no.10 pp. 2782-2793
coculture, crude protein, culture media, diet, enterocytes, essential amino acids, gastric mucosa, human cell lines, jejunum, nitrogen, nutrient uptake, phosphatidylinositol 3-kinase, piglets, protein synthesis, transporters
To investigate effects of dietary nitrogen level on nutrient absorption and utilization in small intestinal enterocyte of piglets, weaned piglets were fed for 10 days with diets containing 20%, 17%, or 14% crude protein (CP) with supplementation to meet requirements for essential amino acids in vivo, and IPEC-1 cells were cultured with different nitrogen levels (NL) in a culture medium (70%, 85%, and 100%) in vitro by monocultured and cocultured intestinal porcine epithelial cells (IPEC-1) and human gastric epithelial cells (GES-1). The results showed the following: (1) In animal trial, decreased dietary CP reduced transcript abundance of nutrient transporters like CAT1, PepT1, GLUT2, and SGLT-1 in jejunal mucosa (0.09 ± 0.03, P < 0.0001; 0.40 ± 0.04, P = 0.0087; 0.20 ± 0.07, P = 0.0003; 0.35 ± 0.02, P = 0.0001), but 17% CP diet did not affect jejunal protein synthesis. (2) The transcript abundance of nutrient transporters displayed similarly effective tendency in jejunal mucosa and cocultured IPEC-1 rather than that in monocultured IPEC-1. (3) Decreased nitrogen levels reduced expressive abundance of PI3K, Class 3 PI3K, TSC2, and 4E-BP1 in monocultured IPEC-1, but 85% nitrogen level did not affect expressive abundance of PI3K, TSC2, mTORC1, 4E-BP1, and S6K1 in cocultured IPEC-1. In general, decreased 3% CP or 15% nitrogen level reduced relative transcript expression of nutrient transporters, but did not affect protein synthesis in jejunal mucosa and cocultured IPEC-1. Therefore, decreased 3% dietary CP increased utilized and synthetic efficiency of nitrogen resource in small intestine and was beneficial in saving the dietary nitrogen resource.