Zhao Bao, Fan Yueli, Li Huijuan, Zhang Chun, Han Rui, Che Dongsheng
Jilin Provincial Key Lab of Animal Nutrition and Feed Science, Key Lab of Animal Production & Product Quality, and Security, Ministry of Education, Jilin Provincial Swine Industry Technical Innovation Center, College of Animal Science and Technology, Jilin Agricultural University, Changchun, China.
College of Animal Science and Technology, Changchun University of Science and Technology, Changchun, China.
J Anim Physiol Anim Nutr (Berl). 2022 May;106(3):664-670. doi: 10.1111/jpn.13677. Epub 2022 Jan 10.
Soybean agglutinin (SBA) is an anti-nutritional factor which decreases the mechanical barrier function in intestinal porcine jejunum epithelial cells (IPEC-J2). Eleutheroside E (EE) is a key part of Acanthopanax senticosus to exert pharmacological effects. This study aims to investigate the effects of EE on the barrier function in IPEC-J2 cells and to determine the ability of EE to enhance the protective effect of barrier function against SBA exposure. The IPEC-J2 cells were cultured in mediums with concentration of 0.1 mg/ml EE, 0.5 ml/ml SBA and 0.1 mg/ml EE pre-treated then treated with 0.5 mg/ml SBA. Then, the transepithelial electric resistance (TEER) value, inflammatory cytokines mRNA expression, tight junction mRNA and protein expression were tested by epithelial Voltohm meter, q-PCR and Western blot method respectively. The results showed that cells treated with 0.1 mg/ml EE had lower permeability (p < 0.05) while 0.5 mg/ml SBA treatment had higher permeability through tested TEER, and higher tight junction proteins (Claudin-3 and ZO-1) expressions and genes (Claudin-3, Occludin and ZO-1) expressions (p < 0.05) in 0.1 mg/ml EE group. IPEC-J2 cells pre-treated with 0.1 mg/ml EE could significantly improve the inflammatory response caused by 0.5 mg/ml SBA by up-regulation for IL-10, TGF-β, and down-regulation gene expression of IL-6, TNF-α and IFN-γ (p < 0.05). In conclusion, 0.1 mg/ml EE can improve the mechanical barrier function and could protect the effects while 0.5 mg/ml of SBA-induced barrier dysfunction in IPEC-J2.
大豆凝集素（SBA）是一种抗营养因子，它会降低猪空肠上皮细胞（IPEC-J2）的机械屏障功能。刺五加苷E（EE）是刺五加发挥药理作用的关键成分。本研究旨在探讨EE对IPEC-J2细胞屏障功能的影响，并确定EE增强屏障功能对SBA暴露的保护作用的能力。将IPEC-J2细胞培养于含有0.1mg/ml EE、0.5mg/ml SBA的培养基中，以及先用0.1mg/ml EE预处理然后再用0.5mg/ml SBA处理的培养基中。然后，分别通过上皮伏特计、q-PCR和蛋白质印迹法检测跨上皮电阻（TEER）值、炎性细胞因子mRNA表达、紧密连接mRNA和蛋白质表达。结果显示，用0.1mg/ml EE处理的细胞通透性较低（p<0.05），而通过检测TEER发现，0.5mg/ml SBA处理的细胞通透性较高，且在0.1mg/ml EE组中紧密连接蛋白（Claudin-3和ZO-1）表达及基因（Claudin-3、闭合蛋白和ZO-1）表达较高（p<0.05）。用0.1mg/ml EE预处理的IPEC-J2细胞可通过上调IL-10、TGF-β以及下调IL-6、TNF-α和IFN-γ的基因表达，显著改善0.5mg/ml SBA引起的炎症反应（p<0.05）。总之，0.1mg/ml EE可改善机械屏障功能，并能保护IPEC-J2细胞免受0.5mg/ml SBA诱导的屏障功能障碍的影响。
