甘草多糖对H_(2)O_(2)诱导肉鸡肝细胞氧化应激损伤的保护作用

Protective effect of glycyrrhiza polysaccharide on H_(2)O_(2)-induced oxidative stress damage in broiler liver cells

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H_(2)O_(2)诱导组、甘草多糖组、H_(2)O_(2)+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H_(2)O_(2)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H_(2)O_(2)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。

The purpose of this study was to explore the protective effect of glycyrrhiza polysaccharide 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,H_(2)O_(2)induction group,glycyrrhiza polysaccharide group and H_(2)O_(2)+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 H_(2)O_(2)induced group,glycyrrhiza polysaccharide pretreatment could significantly enhance liver cell activity and reduce cell apoptosis in broiler chickens;compared with the H_(2)O_(2)induced group,glycyrrhiza polysaccharide pretreatment could significantly increase cell GSH-Px,SOD,CAT,MDA content and T-AOC levels,and inhibit MDA generation;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 reducing MyD88,NF-κB,TNF-α,IL-6 and IL-1β.The results indicated that glycyrrhiza polysaccharide enhance the activity of antioxidant enzymes by activating the Nrf2 signaling pathway and inhibit TLR4 signaling pathway transduction,significantly improving oxidative stress damage in broiler liver cells.