白藜芦醇对热应激诱导的山羊小肠上皮细胞炎性反应调节作用的研究  

Regulatory Effect of Resveratrol on Inflammatory Response Induced by Heat Stress in Goat Small Intestinal Epithelial Cells

作　　者：张宇[1] 徐子洁 黄晓瑜 邢晓南 张小强 赵雷云 张恩平[1] ZHANG Yu;XU Zijie;HUANG Xiaoyu;XING Xiaonan;ZHANG Xiaoqiang;ZHAO Leiyun;ZHANG Enping(College of Animal Science and Technology,Northwest A&F University,Yangling 712100,China)

机构地区：[1]西北农林科技大学动物科技学院,杨凌712100

出　　处：《畜牧兽医学报》2020年第8期1886-1894,共9页ACTA VETERINARIA ET ZOOTECHNICA SINICA

基　　金：国家重点研发计划项目(2016YFD0500508);宁夏自治区重点研发计划项目(2019BBF02001);陕西省重点研发计划项目(2017TSCXL-NY-04-02,2018ZDXM-NY-042)。

摘　　要：本试验旨在探究白藜芦醇对热应激(HS)诱导的山羊小肠上皮细胞炎性因子转录的影响。采用生长良好的第4代山羊小肠上皮细胞(GIE细胞),分为空白对照组(37℃)、热应激组(42℃)、42℃热应激加白藜芦醇组(5、15、30μmol·L-1);加药预处理6 h后进行热应激处理6 h。通过CCK-8法检测白藜芦醇细胞毒性;采用酶联免疫吸附试验(ELISA)检测白藜芦醇对热应激诱导的细胞因子分泌的影响;采用生化法检测白藜芦醇对热应激诱导的GIE细胞抗氧化性能的影响;利用实时荧光定量PCR(RT-PCR)检测白藜芦醇对热应激诱导的GIE细胞HSP70、IL-1β、TNF-α、IL-8和NF-κB基因转录的影响。结果显示:不同浓度(5、15、30μmol·L-1)的白藜芦醇可不同程度地提高热应激诱导的GIE细胞GSH-Px、SOD及T-AOC活性,极显著降低脂质过氧化物MDA含量(P<0.01);不同浓度(5、15、30μmol·L-1)的白藜芦醇能不同程度地抑制热应激诱导的GIE细胞IL-1β、IL-2、IFN-γ和TNF-α的分泌,并极显著降低炎性模型细胞IL-1β、TNF-α、IL-8、HSP 70和NF-κB的基因转录水平(P<0.01)。结果提示,白藜芦醇可调节热应激诱导GIE细胞炎性模型中HSP70基因表达和炎症因子的产生及表达,促进抗氧化酶活力,具有显著的抗炎活性,其抑炎作用可能与抑制NF-κB信号通路相关。The aim of this study was to investigate the effect of resveratrol on the transcription of inflammatory factors in heat stress(HS)-induced goat small intestinal epithelial cells(GIE cells).The fourth-generation well-growing GIE cells were used for the experiment and were divided into five groups:blank control group(37℃),heat stress group(42℃),and 42℃heat stress adding resveratrol groups(5,15,30μmol·L-1);6 hours’heat stress treatment was carried out after 6 hours’pretreatment with resveratrol supplementation.The resveratrol cytotoxicity was detected by CCK-8 method;the effect of resveratrol on the secretion of cytokines was measured by ELISA kits in HS-induced GIE cells;biochemical method was adopted to detect the effect of resveratrol on antioxidant properties of HS-induced GIE cells.Gene transcriptions of IL-1β,TNF-α,IL-8,HSP 70 and NF-κB were detected by RT-PCR in HS-induced GIE cells,respectively.The results showed that resveratrol with different concentrations(5,15,30μmol·L-1)could increase GSH-Px,SOD and T-AOC activities in HS-induced GIE cells,and extremely significantly decrease MDA content of lipid peroxide(P<0.01);inhibit the secretion of IL-1β,IL-2,IFN-γand TNF-αin GIE cells induced by heat stress,and extremely significantly reduce the gene transcription levels of IL-1β,TNF-α,IL-8,HSP 70 and NF-κB in inflammatory model cells(P<0.01).These results demonstrated that resveratrol could regulate the expression of HSP 70 gene and the production and expression of inflammatory factors in the inflammatory model of GIE cells induced by heat stress,promote the activity of antioxidant enzymes and have significant anti-inflammatory activities.Its anti-inflammatory effect might be related to the inhibition of NF-κB signaling pathway.

关 键 词：白藜芦醇 山羊小肠上皮细胞 热应激 HSP70 抗炎 NF-ΚB 

分 类 号：S827[农业科学—畜牧学]