槲皮素通过Nrf2信号通路缓解LPS诱导的牛乳腺上皮细胞氧化应激  

Quercetin alleviates LPS-induced oxidative stress in bovine mammary epithelial cells through the Nrf2 signaling pathway

作　　者：任晓丽 皇甫和平[1] 米俊宪[1] 宋予震[1] 靳双星[1] 王俊东 石冬梅[1] REN Xiaoli;HUANG FU Heping;MI Junxian;SONG Yuzhen;JIN Shuangxing;WANG Jundong;SHI Dongmei(Zhengzhou Key Laboratory of Animal Nutritional Metabolic and Poisoning Diseases,College of Veterinary Medicine,Henan University of Animal Husbandry of Economy,Zhengzhou 450046,China)

机构地区：[1]河南牧业经济学院动物医药学院,郑州市动物营养代谢病与中毒病重点实验室,河南郑州450046

出　　处：《中国兽医学报》2023年第12期2590-2596,共7页Chinese Journal of Veterinary Science

基　　金：河南省科技攻关资助项目(222102110110,212102110377);河南牧业经济学院校级重点学科建设资助项目(XJXK202202)。

摘　　要：旨在探究槲皮素(quercetin,QU)通过Nrf2信号通路对脂多糖(lipopolysaccharide,LPS)诱导的牛乳腺上皮细胞氧化应激的作用及其机制。以牛乳腺上皮细胞系MAC-T细胞为研究对象,用2.5、5.0和10.0μmol/L QU预处理MAC-T细胞12 h后加入1 mg/L LPS处理12 h后;利用试剂盒分别检测LDH活性、活性氧(reactive oxygen species,ROS)、脂质过氧化物丙二醛(malondialdehyde,MDA)的含量、超氧化物歧化酶(superoxide dismutase,SOD)、谷胱甘肽过氧物酶(glutathione peroxidase,GSH-Px)、过氧化氢酶(catalase,CAT)活性。Western blot和RT-qPCR方法分别检测核转录因子Nrf2相关因子(nuclear factor E2-related factor,Nrf2)、血红素加氧酶1(heme oxygenase-1,HO-1)和醌氧化还原酶NADH1(NADPH quinone oxidoreductase 1,NQO1)基因和蛋白表达变化。结果显示,与对照组相比,1 mg/L LPS组中ROS和MDA的含量显著升高(P<0.05),SOD、GSH-Px和CAT活性显著降低(P<0.05),Nrf2、HO-1和NQO1基因和蛋白相对表达水平显著降低(P<0.05);与LPS组相比,QU预处理能显著降低ROS和MDA的含量(P<0.05),而SOD和GSH-px活性显著升高,且呈剂量依赖性;此外,QU+LPS组能显著上调MAC-T细胞内Nrf2、HO-1和NQO1基因和蛋白相对表达水平(P<0.05)。结果表明,QU通过激活Nrf2信号通路缓解LPS诱导的牛乳腺上皮细胞的氧化应激。To investigate the effect and mechanism of quercetin(QU)on lipopolysaccharide(LPS)-induced oxidative stress in bovine mammary epithelial cells through nuclear factor E2-reated factor2(Nrf2)signaling pathway.The bovine mammary epithelial cells line MAC-T cells were pretreated with different concentrations of QU(2.5,5.0 and 10.0μmol/L)for 12 h,and then treated with1 mg/L LPS for 12 h.The kits were used to detect the activity of LDH,ROS level,MDA content,and the activity of SOD and GSH-Px.qRT-PCR and Western blot method were used to detect the relative expression of Nrf2,HO-1 and NQO1 mRNA and protein.The results showed that,compared with the control group,the content of ROS and MDA in LPS group were significantly increased(P<0.05),while the activity of SOD and GSH-Px were significantly decreased(P<0.05),the relative expression levels of Nrf2,HO-1 and NQO1 genes and proteins were significantly decreased;compared with LPS group,QU pretreatment significantly reduced the contents of ROS and MDA(P<0.05),while the activities of SOD and GSH-px were significantly increased in a dose-dependent manner.Moreover,QU pretreatment obviously increased the relative expression levels of Nrf2,HO-1 and NQO1 mRNA and protein in MAC-T cells(P<0.05).These results suggested that QU alleviated LPS-induced oxidative stress in bovine mammary epithelial cells by activating the Nrf2 signaling pathway.

关 键 词：槲皮素 LPS MAC-T细胞 氧化应激 Nrf2信号通路 

分 类 号：S856.9[农业科学—临床兽医学]