Li Yunyun, Lu Xiaoxi, Wu Haiqin, Xia Mi, Hou Qihang, Hu Weiwei, Li Tiejun, Wu Li, Yu Qinghua
MOE Joint International Research Laboratory of Animal Health and Food Safety, College of Veterinary Medicine, Nanjing Agricultural University, Weigang 1, Nanjing, Jiangsu 210095, PR China.
Institute of Subtropical Agriculture, Chinese Academy of Sciences, Changsha, Hunan 410125, PR China.
Res Vet Sci. 2019 Feb;122:15-21. doi: 10.1016/j.rvsc.2018.11.013. Epub 2018 Nov 12.
To explore the effects of reducing the Cp levels on intestinal barrier function, low Cp (LP) and NRC standard Cp (NP) diets were fed to pigs from 45 to 160 days, and in vitro experiments were performed using monolayers of IPEC-J2 cells. The number of goblet cells, expression of proteins related to cell junction, amino acid transport, glucose transport, transepithelial electrical resistance (TEER), dextran permeability, and IL-6 secretion level were detected in pigs. The results demonstrated that a moderate reduction of Cp levels did not affect intestinal morphology, as demonstrated by a normal villi height, crypt depth and normal numbers of goblet cells. The maintenance of the intestinal structure obtained with LP was also confirmed by stable mRNA expression levels of muc2 and E-cadherin in the jejunum. We also found that LP did not affect the protein expression of cationic amino acid transporter 1 (CAT-1) and alanine serine cysteine transporter 1 (ASCT1) from 45 to 160 days. Moreover, the excitatory amino acid transporter 3 (EAAT3), sodium-glucose cotransporter 1 (SGLT1) and glucose transporter (GLUT2) protein expression levels in the jejunum were significantly increased at a certain age during the rearing period. Furthermore, we also demonstrated that a reduction in protein concentration up to 15% in the cultural medium of IPEC-J2 cells did not impact the mucosal barrier function. This study demonstrated that a moderate reduction of the protein level did not affect intestinal mucosal barrier function and morphology in the jejunum.
为探究降低铜(Cp)水平对肠道屏障功能的影响,从45日龄至160日龄给猪饲喂低铜(LP)日粮和NRC标准铜(NP)日粮,并使用IPEC-J2细胞单层进行体外实验。检测了猪的杯状细胞数量、与细胞连接、氨基酸转运、葡萄糖转运、跨上皮电阻(TEER)、葡聚糖通透性及白细胞介素-6分泌水平相关的蛋白质表达。结果表明,适度降低铜水平不影响肠道形态,表现为绒毛高度、隐窝深度正常,杯状细胞数量正常。空肠中muc2和E-钙黏蛋白的mRNA表达水平稳定也证实了LP日粮维持了肠道结构。我们还发现,从45日龄至160日龄,LP日粮不影响阳离子氨基酸转运体1(CAT-1)和丙氨酸丝氨酸半胱氨酸转运体1(ASCT1)的蛋白质表达。此外,在饲养期的特定年龄段,空肠中兴奋性氨基酸转运体3(EAAT3)、钠-葡萄糖协同转运蛋白1(SGLT1)和葡萄糖转运蛋白(GLUT2)的蛋白质表达水平显著升高。此外,我们还证明,将IPEC-J2细胞培养基中的蛋白质浓度降低至15%不会影响黏膜屏障功能。本研究表明,适度降低蛋白质水平不会影响空肠的肠道黏膜屏障功能和形态。