关键词: 大豆球蛋白, 猪小肠上皮细胞, 信号通路

Abstract: In this study, porcine intestinal epithelial cells (IPEC-J2) were cultured in vitro to study the effect of different concentrations of glycinin (11S) on IPEC-J2. IPEC-J2 cells were divided into six groups. A was control group, in groups B (1 mg·mL^-1 11S), C (5 mg·mL^-1 11S), D (10 mg·mL^-1 11S), E (5 mg·mL^-1 11S+1 μmol·L^-1 PDTC) and F (5 mg·mL^-1 11S+1 μmol·mL^-1 L-NAME), 0, 1, 5, 10, 5, and 5 mg·mL^-1 of 11S were added respectively. In addition, 1 μmol·L^-1 PDTC and 1 μmol·mL^-1 L-NAME were added to groups E and F, respectively. After 8, 16 and 24 h, cell viability was measured by CCK-8 method, contents of NO, DAO, 5-HT, IL-6 and IL-10 were detected by ELISA, and p-NF-κB, p65, iNOS and COX-2 protein expression levels were detected by western blot, relative expression of NF-κB p65, IKKβ, iNOS, IKKα and COX-2 mRNA were determined by PCR. The results showed that 11S reduced viability of porcine intestinal epithelial cells, promoted the secretion of NO, 5-HT and IL-6, decreased the secretion of IL-10, and increased expression of p-NF-κB p65, iNOS and COX-2 proteins, and expression of NF-κB p65, IKKβ, iNOS, IKKα and COX-2 mRNA were also increased. PDTC and L-NAME could inhibit the effects of 11S. The results showed that 11S caused damage to IPEC-J2 and increased the damage with increasing concentration, PDTC and L-NAME could reduce the damage of 11S to cells.

Key words: glycinin, porcine intestinal epithelial cells, signaling pathway