摘要
探讨甘草多糖对氧化应激造成的肉鸡肝细胞损伤的保护作用,并对其作用机制进行研究.以体外培养的原代肉鸡肝细胞为空白对照组,同时设置H2O2诱导组、甘草多糖组、H2O2+甘草多糖组,用CCK-8试剂检测肉鸡肝细胞活性,JC-1和DAPI染色试剂盒检测细胞凋亡,相关试剂盒检测GSH-Px、SOD、CAT、MDA含量和T-AOC水平,qRT-PCR检测肉鸡肝细胞中抗氧化基因Nrf2、Keap-1、GPx1、SOD、CAT和炎症基因TLR4、MyD88、NF-κB、TNF-α、IL-6、IL-1β、IL-4、IL-10的相对表达量.结果表明,与H2O2诱导组相比,甘草多糖预处理可明显提升肉鸡肝细胞活性,减少细胞凋亡;与H2O2诱导组相比,甘草多糖预处理可明显提高细胞GSH-Px、SOD、CAT、MDA含量和T-AOC水平,抑制MDA生成;qRT-PCR检测显示,甘草多糖可显著提高Nrf2、Keap-1、GPx1、SOD、CAT、IL-4、IL-10的相对表达量,同时显著降低MyD88、NF-κB、TNF-α、IL-6、IL-1β的相对表达量.甘草多糖可能通过激活Nrf2信号通路提高抗氧化酶的活性,并抑制TLR4信号通路,从而明显改善肉鸡肝细胞的氧化应激损伤.
Abstract
The purpose of this study was to explore the protective effect of glycyrrhiza polysaccha-ride on oxidative stress-induced liver cell damage in broiler chickens,and to study its mechanism of action.Primary broiler liver cells cultured in vitro were divided into blank control group,H2 O2 in-duction group,glycyrrhiza polysaccharide group and H2O2+glycyrrhiza polysaccharide group;CCK-8 reagent was used to detect liver cell activity in broiler chickens,JC-1 and DAPI staining kits were used to detect cell apoptosis,relevant kits were used to detect GSH-Px,SOD,CAT,MDA content,and T-AOC levels.qRT-PCR was used to detect antioxidant genes Nrf2,Keap-1,GPx1,SOD,CAT and inflammatory genes TLR4,MyD88,NF-κB,TNF-α、IL-6,IL-1β、IL-4 and IL-10.The results showed that compared with the H2O2 induced group,glycyrrhiza polysaccharide pretreat-ment could significantly enhance liver cell activity and reduce cell apoptosis in broiler chickens;compared with the H2O2 induced group,glycyrrhiza polysaccharide pretreatment could significant-ly increase cell GSH-Px,SOD,CAT,MDA content and T-AOC levels,and inhibit MDA genera-tion;qRT-PCR detection showed that glycyrrhiza polysaccharide could significantly increase the relative expression levels of Nrf2,Keap-1,GPx1,SOD,CAT,IL-4 and IL-10,while significantly re-ducing MyD88,NF-κB,TNF-α,IL-6 and IL-1β.The results indicated that glycyrrhiza polysaccha-ride enhance the activity of antioxidant enzymes by activating the Nrf2 signaling pathway and in-hibit TLR4 signaling pathway transduction,significantly improving oxidative stress damage in broiler liver cells.
基金项目
国家自然科学基金资助项目(31872537)
国家重点研发计划资助项目(2022YFE0111100)
万方数据