College of Animal Science and Technology, Jilin Agricultural University, Changchun 130118, China.
Int J Mol Sci. 2013 Nov 1;14(11):21689-704. doi: 10.3390/ijms141121689.
In this study, we sought to investigate the role of soybean agglutinin (SBA) in mediating membrane permeability and the mechanical barrier function of intestinal epithelial cells. The IPEC-J2 cells were cultured and treated with 0, 0.5, 1.0, 1.5, 2.0, 2.5, or 3.0 mg/mL SBA. Transepithelial electrical resistance (TEER) and alkaline phosphatase (AP) activity were measured to evaluate membrane permeability. The results showed a significant decrease in TEER values (p < 0.05) in a time- and dose-dependent manner, and a pronounced increase in AP activity (p < 0.05). Cell growth and cell morphology were used to evaluate the cell viability. A significant cell growth inhibition (p < 0.05) and alteration of morphology were observed when the concentration of SBA was increased. The results of western blotting showed that the expression levels of occludin and claudin-3 were decreased by 31% and 64% compared to those of the control, respectively (p < 0.05). In addition, immunofluorescence labeling indicated an obvious decrease in staining of these targets and changes in their localizations. In conclusion, SBA increased the membrane permeability, inhibited the cell viability and reduced the levels of tight junction proteins (occludin and claudin-3), leading to a decrease in mechanical barrier function in intestinal epithelial cells.
在这项研究中,我们试图探讨大豆凝集素(SBA)在介导肠上皮细胞的膜通透性和机械屏障功能中的作用。我们培养了 IPEC-J2 细胞,并分别用 0、0.5、1.0、1.5、2.0、2.5 或 3.0mg/mL 的 SBA 处理这些细胞。我们测量了跨上皮电阻(TEER)和碱性磷酸酶(AP)活性,以评估膜通透性。结果表明,TEER 值呈时间和剂量依赖性显著降低(p<0.05),AP 活性显著增加(p<0.05)。细胞生长和细胞形态用于评估细胞活力。当 SBA 浓度增加时,观察到明显的细胞生长抑制(p<0.05)和形态改变。Western blot 结果表明,与对照组相比,occludin 和 claudin-3 的表达水平分别降低了 31%和 64%(p<0.05)。此外,免疫荧光标记表明这些靶标的染色明显减少,其定位也发生了变化。总之,SBA 增加了膜通透性,抑制了细胞活力,并降低了紧密连接蛋白(occludin 和 claudin-3)的水平,导致肠上皮细胞的机械屏障功能下降。