血根碱通过核转录因子-κB信号通路减轻脂多糖诱导的仔猪小肠黏膜上皮细胞炎症反应  

Sanguinarine Inhibits Inflammatory Response of Piglet Small Intestinal Mucosal Epithelial Cells Induced by Lipopolysaccharide through Nuclear Factor-κB Signaling Pathway

作　　者：高欢 蒋晓旭 杨玉婷 冉津铭 沈丽艳 蒋恒鑫 李雪艳 林秋叶[1] 蒋庆文 曹振辉[1] 潘洪彬[1] GAO Huan;JIANG Xiaoxu;YANG Yuting;RAN Jinming;SHEN Liyan;JIANG Hengxin;LI Xueyan;LIN Qiuye;JIANG Qingwen;CAO Zhenhui;PAN Hongbin(Yunnan Key Laboratory of Animal Nutrition and Feed,College of Animal Science and Technology,Yunnan Agricultural University,Kunming 650201,China;Dazhou Vocational and Technical College,Dazhou 635000,China;Institute of Spices Yunnan Agricultural University,Kunming 650201,China)

机构地区：[1]云南农业大学动物科学技术学院,云南省动物营养重点实验室,昆明650201 [2]达州职业技术学院,达州635000 [3]云南农业大学香料研究所,昆明650201

出　　处：《动物营养学报》2021年第3期1675-1686,共12页CHINESE JOURNAL OF ANIMAL NUTRITION

基　　金：国家十三五重点研发计划项目(2018YFD0500401);云南省科技计划项目(2018FA021);国家自然科学基金委员会(U1802234,31760645)。

摘　　要：本试验旨在研究血根碱(SG)通过核转录因子-κB(NF-κB)信号通路调控仔猪小肠黏膜上皮细胞(IPEC-J2细胞)抗炎作用机制。在探明SG对IPEC-J2细胞促生长作用的适宜浓度和时间以及构建IPEC-J2细胞致炎模型的LPS适宜浓度和时间的基础上,检测对照组(无添加)、致炎模型细胞组(LPS组,5.0μg/mL LPS处理1 h)和适宜浓度SG+LPS致炎模型细胞组(SG+LPS组,5.0μg/mL LPS处理1 h和0.500μg/mL SG处理24 h)细胞中NF-κB信号通路相关基因和蛋白表达及其产物含量。结果表明:1)SG对IPEC-J2细胞促生长作用的适宜浓度和时间分别为0.500μg/mL和24 h,构建IPEC-J2细胞致炎模型LPS的适宜浓度和时间分别为5.0μg/mL和1 h。2)与对照组相比,LPS组白细胞介素-6(IL-6)、白细胞介素-8(IL-8)、核转录因子-κB亚基1(NF-κB1A)、核转录因子-κB2(NF-κB2)、肿瘤坏死因子-α诱导蛋白3(TNFAIP3)、核转录因子κB激酶抑制剂ε(IKKε)、核转录因子-κB1(NF-κB1)、RelB mRNA相对表达量均极显著升高(P<0.01);细胞上清液中IL-6、IL-8和肿瘤坏死因子-α(TNF-α)含量显著或极显著升高(P<0.05或P<0.01);细胞总蛋白中核转录因子-κB抑制蛋白α(IκBα)、p65蛋白表达量极显著降低(P<0.01),核蛋白中p65蛋白表达量极显著升高(P<0.01),胞浆蛋白中p65蛋白表达量极显著下降(P<0.01)。3)与LPS组相比,SG+LPS组细胞中IL-6、IL-8、NF-кB1A、NF-кB2、TNFAIP3、IKKε、NF-кB1 mRNA相对表达量显著或极显著降低(P<0.05或P<0.01);细胞上清液中IL-6和IL-8含量显著或极显著降低(P<0.05或P<0.01);细胞总蛋白中IκBα蛋白表达量极显著升高(P<0.01),核蛋白中p65蛋白表达量极显著下降(P<0.01),胞浆蛋白中p65蛋白表达量极显著升高(P<0.01)。由此可见,SG通过抑制NF-κB信号通路相关基因的活化和p65核转移,减轻IPEC-J2细胞的炎症反应。This study was to elucidate the anti-inflammatory regulate mechanism of sanguinarine(SG)by nuclear factor-κB(NF-κB)signaling pathway in piglet small intestinal mucosal epithelial cells(IPEC-J2 cells).In this study,we investigated the appropriate concentration and time for SG to promote the growth of IPEC-J2 cells and the appropriate concentration and time for LPS to establish an inflammatory model of IPEC-J2 cells.The expression of the key genes and protein expression and product content in NF-κB signal pathway among the control group(no added),inflammation model cell group(LPS group,5.0μg/mL LPS treatment for 1 h)and appropriate concentration of SG+LPS-induced inflammation model cell group(SG+LPS group,5.0μg/mL LPS treatment for 1 h and 0.500μg/mL SG treatment for 24 h)were detected.The results showed as follows:1)the appropriate concentration and time for SG to promote the growth of IPEC-J2 cells were 0.500μg/mL and 24 h,respectively,and the appropriate concentration and time for the construction of IPEC-J2 cell inflammatory model of LPS were 5.0μg/mL and 1 h,respectively.2)Compared with the control group,the mRNA relative expression levels of interleukin-6(IL-6),interleukin-8(IL-8),nuclear factor-κB subunit 1(NF-κB1A),nuclear factor-κB2(NF-κB2),tumor necrosis factor-αinducible protein 3(TNFAIP3),nuclear factor-κB kinase inhibitorsε(IKKε),nuclear factor-κB1(NF-κB1)and RelB in the LPS group were significantly increased(P<0.01);the contents of IL-6,IL-8 and TNF-αin cell supernatant were significantly increased(P<0.05 or P<0.01);the protein expression levels of nuclear factor-κB inhibitor proteinα(IκBα)and p65 in cells total protein were significantly decreased(P<0.05),the nucleoprotein p65 protein expression level was significantly increased(P<0.01),and the cytoplasmic protein p65 protein expression level was significantly decreased(P<0.01).3)Compared with the LPS group,the mRNA relative expression levels of IL-6,IL-8,NF-κB1A,NF-κB2,TNFAIP3,IKKεand NF-κB1 in the SG+LPS group were signif

关 键 词：血根碱 IPEC-J2细胞 NF-ΚB信号通路 抗炎 

分 类 号：S826[农业科学—畜牧学]