Jilin Provincial Key Lab of Animal Nutrition and Feed Science, Jilin Agricultural University, Changchun, China.
College of Animal Science and Technology, Jilin Agricultural University, Changchun, China.
J Anim Physiol Anim Nutr (Berl). 2019 Jul;103(4):1174-1184. doi: 10.1111/jpn.13087. Epub 2019 Apr 16.
Eleutheroside B (EB) is a phenylpropanoid glycoside with anti-inflammatory properties, neuroprotective abilities, immunomodulatory effects, antinociceptive effects, and regulation of blood glucose. The aim of this study was to investigate the effects of EB on the barrier function in the intestinal porcine epithelial cells J2 (IPEC-J2). The IPEC-J2 cells were inoculated into 96-well plates at a density of 5 × 10 cells per well for 100% confluence. The cells were cultured in the presence of EB at concentrations of 0, 0.05, 0.10, and 0.20 mg/ml for 48 hr. Then, 0.10 mg/ml was selected as the suitable concentration for the estimation of transepithelial electric resistance (TEER) value, alkaline phosphatase activity, proinflammatory cytokines mRNA expression, tight junction mRNA and protein expression. The results of this study indicated that the supplementation of EB in IPEC-J2 cells decreased cellular membrane permeability and mRNA expression of proinflammatory cytokines, including interleukin-6 (IL-6), interferon-γ (INF-γ), and tumour necrosis factor-α (TNF-α). The supplementation of EB in IPEC-J2 cells increased tight junction protein expression and anti-inflammatory cytokines, interleukin 10 (IL-10) and transforming growth factor beta (TGF-β). In addition, the western blotting and real-time quantitative polymerase chain reaction (RT-qPCR) results indicated that EB significantly (p < 0.05) increased the mRNA and protein expression of intestinal tight junction proteins, Claudin-3, Occludin, and Zonula Occludins protein-1 (ZO-1). Therefore, dietary supplementation of EB may increase intestinal barrier function, tight junction protein expression, anti-inflammatory cytokines, and decrease proinflammatory cytokines synthesis in IPEC-J2 cells.
淫羊藿苷 B (EB) 是一种具有抗炎、神经保护、免疫调节、镇痛和调节血糖作用的苯丙素糖苷。本研究旨在探讨 EB 对肠道猪上皮细胞 J2 (IPEC-J2) 屏障功能的影响。将 IPEC-J2 细胞以 5×10 个/孔的密度接种于 96 孔板中，待细胞达到 100%汇合度时进行培养。将细胞分别在 0、0.05、0.10 和 0.20 mg/ml 的 EB 浓度下培养 48 小时。然后，选择 0.10 mg/ml 作为估计跨上皮电阻 (TEER) 值、碱性磷酸酶活性、促炎细胞因子 mRNA 表达、紧密连接 mRNA 和蛋白表达的适宜浓度。结果表明，EB 补充到 IPEC-J2 细胞中降低了细胞通透性和促炎细胞因子（包括白细胞介素 6 (IL-6)、干扰素-γ (INF-γ) 和肿瘤坏死因子-α (TNF-α)）的 mRNA 表达。EB 补充到 IPEC-J2 细胞中增加了紧密连接蛋白的表达和抗炎细胞因子白细胞介素 10 (IL-10) 和转化生长因子-β (TGF-β)。此外，Western blot 和实时定量聚合酶链反应 (RT-qPCR) 结果表明，EB 显著（p<0.05）增加了肠道紧密连接蛋白 Claudin-3、Occludin 和 Zonula Occludins 蛋白-1 (ZO-1) 的 mRNA 和蛋白表达。因此，膳食补充 EB 可能会增加 IPEC-J2 细胞的肠道屏障功能、紧密连接蛋白表达、抗炎细胞因子，并减少促炎细胞因子的合成。
