Jilin Provincial Key Laboratory of Animal Nutrition and Feed Science, Jilin Agricultural University, Changchun, China.
Department of Animal Science and Technology, Jilin Agricultural University, Changchun, China.
J Anim Physiol Anim Nutr (Berl). 2019 Jul;103(4):1198-1206. doi: 10.1111/jpn.13091. Epub 2019 Apr 1.
Soya bean agglutinin (SBA) is a glycoprotein and the main anti-nutritional component in most soya bean feedstuffs. It is mainly a non-fibre carbohydrate-based protein and represents about 10% of soya bean-based anti-nutritional effects. In this study, we sought to determine the effects of N-Acetyl-D-galactosamine (GalNAc or D-GalNAc) on the damage induced by SBA on the membrane permeability and tight junction proteins of piglet intestinal epithelium (IPEC-J2) cells. The IPEC-J2 cells were pre-cultured with 0, 0.125 × 10 , 0.25 × 10 , 0.5 × 10 , 1.0 × 10 and 2.0 × 10 mmol/L GalNAc at different time period (1, 2, 4 and 8 hr) before being exposed to 0.5 mg/ml SBA for 24 hr. The results indicate that pre-incubation with GalNAc mitigates the mechanical barrier injury as reflected by a significant increase in trans-epithelial electric resistance (TEER) value and a decrease in alkaline phosphatase (ALP) activity in cell culture medium pre-treated with GalNAc before incubation with SBA as both indicate a reduction in cellular membrane permeability. In addition, mRNA levels of the tight junction proteins occludin and claudin-3 were lower in the SBA-treated groups without pre-treatment with GalNAc. The mRNA expression of occludin was reduced by 17.3% and claudin-3 by 42% (p < 0.01). Moreover, the corresponding protein expression levels were lowered by 17.8% and 43.5% (p < 0.05) respectively. However, in the GalNAc pre-treated groups, occludin and claudin-3 mRNAs were reduced by 1.6% (p > 0.05) and 2.7% (p < 0.01), respectively, while the corresponding proteins were reduced by 4.3% and 7.2% (p < 0.05). In conclusion, GalNAc may prevent the effect of SBA on membrane permeability and tight junction proteins on IPEC-J2s.
大豆凝集素（SBA）是一种糖蛋白，也是大多数大豆饲料中的主要抗营养成分。它主要是一种非纤维碳水化合物蛋白，占大豆抗营养作用的约 10%。在这项研究中，我们试图确定 N-乙酰-D-半乳糖胺（GalNAc 或 D-GalNAc）对 SBA 诱导的仔猪肠上皮（IPEC-J2）细胞的膜通透性和紧密连接蛋白损伤的影响。在与 0.5mg/ml SBA 孵育 24 小时之前，将 IPEC-J2 细胞用 0、0.125×10、0.25×10、0.5×10、1.0×10 和 2.0×10mmol/L GalNAc 预培养不同时间（1、2、4 和 8 小时）。结果表明，GalNAc 预处理可减轻机械屏障损伤，表现在用 GalNAc 预处理后，与 SBA 孵育的细胞培养液中转 上皮电阻（TEER）值显著增加，碱性磷酸酶（ALP）活性降低，表明细胞通透性降低。此外，未经 GalNAc 预处理的 SBA 处理组中紧密连接蛋白 occludin 和 claudin-3 的 mRNA 水平较低。occludin 的 mRNA 表达降低了 17.3%，claudin-3 降低了 42%（p<0.01）。此外，相应的蛋白表达水平分别降低了 17.8%和 43.5%（p<0.05）。然而，在 GalNAc 预处理组中，occludin 和 claudin-3 的 mRNA 分别降低了 1.6%（p>0.05）和 2.7%（p<0.01），而相应的蛋白分别降低了 4.3%和 7.2%（p<0.05）。总之，GalNAc 可能预防 SBA 对 IPEC-J2s 膜通透性和紧密连接蛋白的作用。
