7S β-伴大豆球蛋白通过NF-κB信号通路引起IPEC-J2细胞的炎性反应

doi:10.11843/j.issn.0366-6964.2019.04.020

摘要/Abstract

摘要：

采用细胞体外培养技术，研究不同浓度7S β-伴大豆球蛋白对猪小肠上皮细胞（IPEC-J2细胞）的影响。试验分为A、B、C、D、E、F 6组，其中A为对照组，其余各组中分别添加1、5、10、5、5 mg·mL^-1的β-伴大豆球蛋白，并且在E和F组分别添加1 μmol·L^-1 NF-κB（PDTC）和iNOS（L-NAME）抑制剂。用CCK-8法检测各组细胞存活率，用ELISA法检测细胞NO、DAO、5-HT、IL-6和IL-10含量，用Western blot法检测细胞p-NF-κB p65、iNOS、COX-2的蛋白表达量，用qPCR法检测细胞NF-κB p65、IKKβ、iNOS、IKKα、COX-2 mRNA的相对转录量。结果显示：β-伴大豆球蛋白降低IPEC-J2细胞存活率，添加PDTC和L-NAME增高IPEC-J2细胞存活率，促进NO、DAO、5-HT和IL-6的分泌，并降低IL-10的分泌，同时增加p-NF-κB p65、iNOS、COX-2的蛋白表达量及NF-κB p65、IKKβ、iNOS、IKKα、COX-2 mRNA的相对转录量，添加PDTC和L-NAME后抑制了β-伴大豆球蛋白的作用。结果表明，β-伴大豆球蛋白引起IPEC-J2细胞损伤，随浓度增大损伤增加，PDTC和L-NAME可以降低β-伴大豆球蛋白对细胞的作用。

关键词: β-伴大豆球蛋白, IPEC-J2细胞, NF-κB, PDTC, L-NAME

Abstract:

In this study, IPEC-J2 cells (porcine intestinal epithelial cells) were cultured in vitro to study the effect of different concentrations of 7S β-conglycinin on IPEC-J2. The experiment was designed as six groups:A, B, C, D, E, and F, A was the control group, and the other groups were added 1, 5, 10, 5 and 5 mg·mL^-1β-conglycinin, respectively, and 1 μmol·L^-1 NF-κB(PDTC) and iNOS (L-NAME) inhibitors were added to groups E and F, respectively. Cell viability was measured by CCK-8 method, the contents of NO, DAO, 5-HT, IL-6 and IL-10 were detected by ELISA, p-NF-κB, p65, iNOS and COX-2 protein expression levels were detected by Western blot, the relative transcription of NF-κB p65, IKKβ, iNOS, IKKα and COX-2 mRNA were determined by qPCR. The results showed that β-conglycinin reduced the viability of IPEC-J2 cells, promoted the secretion of NO, 5-HT and IL-6, decreased the secretion of IL-10, and increased the expression of p-NF-κB p65, iNOS and COX-2 proteins, transcription of NF-κB p65, IKKβ, iNOS, IKKα and COX-2 mRNA, PDTC and L-NAME can inhibit the effects of β-conglycinin. The results showed that β-conglycinin caused damage to IPEC-J2 and increased the damage with increasing concentration, PDTC and L-NAME can reduce the damage of β-conglycinin to cells.

Key words: β-conglycinin, IPEC-J2, NF-κB, PDTC, L-NAME

中图分类号:

Q251

彭成璐, 张瑜, 丁雪东, 李玉, 冯士彬, 王希春, 李锦春, 吴金节. 7S β-伴大豆球蛋白通过NF-κB信号通路引起IPEC-J2细胞的炎性反应[J]. 畜牧兽医学报, 2019, 50(4): 870-878.

PENG Chenglu, ZHANG Yu, DING Xuedong, LI Yu, FENG Shibin, WANG Xichun, LI Jinchun, WU Jinjie. 7S β-conglycinin Triggered Inflammatory Response in IPEC-J2 Cells through NF-κB Signaling Pathway[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2019, 50(4): 870-878.

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