复配黄豆苷元-藤茶黄酮调控核因子E2相关因子2-Kelch样环氧氯丙烷相关蛋白1信号通路抑制叔丁基过氧化氢引起蛋鸡卵巢颗粒细胞氧化损伤  

Compound Daidzein-Dihydromyricetin Regulates Nuclear Factor E2-Related Factor 2-Kelch-Like Epichlorohydrin-Associated Protein 1 Signaling Pathway and Inhibits 1-Butyl Hydroperoxide Induced Oxidative Damage in Ovarian Granulosa Cells of Laying Hens

作　　者：鄂晓迪 胡友军 赵晓南 程皇座 文伟 施晓丽[2] E Xiaodi;HU Youjun;ZHAO Xiaonan;CHENG Huangzuo;WEN Wei;SHI Xiaoli(Guangdong Nuacid Nutrition Co.,Ltd.,Qingyuan 511500,China;Key Laboratory of Genetic Breeding and Reproduction in the Plateau Mountain Region,Ministry of Education,College of Animal Science,Guizhou University,Guiyang 550025,China)

机构地区：[1]广东酸动力生物科技有限公司,清远511500 [2]高原山地动物遗传育种与繁殖教育部重点实验室,贵州大学动物科学学院,贵阳550025

出　　处：《动物营养学报》2025年第4期2674-2683,共10页CHINESE JOURNAL OF ANIMAL NUTRITION

基　　金：国家重点研发计划(2023YFD1301203-04)。

摘　　要：本试验旨在研究复配黄豆苷元-藤茶黄酮(DA-DHM)是否通过介导核因子E2相关因子2(Nrf2)-Kelch样环氧氯丙烷相关蛋白1(Keap1)信号通路来缓解叔丁基过氧化氢(TBHP)引起的蛋鸡卵巢颗粒细胞(GCs)氧化损伤。以蛋鸡卵巢GCs为研究对象,利用不同浓度TBHP(0、200、400、600、800μmol/L)分别处理细胞12和24 h,利用CCK-8法检测细胞活力,确定TBHP的浓度和时间。试验分为4个组:C组在M199基础培养基处理24 h;T1组在含400μmol/L的TBHP培养基处理12 h;T2组为在含10 ng/mL的DA-DHM培养基处理12 h后,更换含400μmol/L的TBHP培养基处理12 h;T3组在含10μmol/L的ML385培养基处理12 h后,更换含10 ng/mL的DA-DHM培养基处理12 h,再更换含400μmol/L的TBHP培养基处理12 h。检测细胞活性、活性氧(ROS)含量、线粒体膜电位、钙离子(Ca^(2+))浓度以及抗氧化相关基因mRNA的相对表达量。结果表明:1)TBHP的最佳造模剂量和时间分别为400μmol/L和12 h。2)与C组相比,T1组GCs内ROS含量、Ca^(2+)浓度和Keap1的mRNA的相对表达量显著升高(P<0.05),细胞线粒体膜电位和Nrf2、过氧化氢酶(CAT)、超氧化物歧化酶1(SOD1)和谷氨酸半胱氨酸连接酶催化亚基(GCLC)的mRNA的相对表达量显著降低(P<0.05)。与T1组相比,T2组GCs内ROS含量、Ca^(2+)浓度和Keap1的mRNA的相对表达量显著降低(P<0.05),线粒体膜电位和Nrf2、SOD1、谷氨酸半胱氨酸连接酶修饰亚基(GCLM)的mRNA的相对表达量显著升高(P<0.05)。与T2组相比,T3组GCs内Nrf2、CAT、SOD1、GCLM和GCLC的mRNA相对表达量显著降低(P<0.05),Keap1的mRNA相对表达量显著提高(P<0.05)。综上所述,DA-DHM能够缓解TBHP引起的蛋鸡卵巢GCs氧化应激损伤,其潜在机制可能是通过调控Nrf2-Keap1信号通路上下游基因的表达,从而减轻GCs的氧化应激和线粒体损伤,并抑制细胞凋亡。This study aimed to explore whether the compound daidzein-daxanthin(DA-DHM)relieved 1-butyl hydroperoxide(TBHP)induced oxidative damage in ovarian granulosa cells(GCs)of laying hens by mediating nuclear factor E2-related factor 2(Nrf2)-Kelch-like epichlorohydrin-associated protein 1(Keap1)signaling pathway.Taking ovarian GCs of laying hens as the research subjects,different concentrations of TBHP(0,200,400,600 and 800μmol/L)were used to treat cells for 12 and 24 h,respectively.The cell viability was detected using the CCK-8 method,to determine the concentration and time of TBHP.The experiment was divided into 4 groups:group C was treated with M199 basal medium for 24 h;group T1 was treated with medium containing 400μmol/L TBHP for 12 h;group T2 was treated with medium containing 10 ng/mL DA-DHM for 12 h,and the medium containing 400μmol/L TBHP was replaced for 12 h;group T3 was treated with medium containing 10μmol/L ML385 for 12 h,and the medium containing 10 ng/mL DA-DHM was replaced for 12 h,and the medium containing 400μmol/L TBHP was replaced again for 12 h.The cell viability,reactive oxygen species(ROS)content,mitochondrial membrane potential,calcium ion(Ca^(2+))concentration and mRNA relative expression levels of antioxidant related genes were detected.The results showed as follows:1)the optimal modeling dose and time for TBHP were 400μmol/L and 12 h,respectively.2)Compared with group C,the ROS content,Ca^(2+)concentration and Keap1 mRNA relative expression level in GCs of group T1 were significantly increased(P<0.05),and the mitochondrial membrane potential and mRNA relative expression levels of Nrf2,catalase(CAT),superoxide dismutase 1(SOD1)and glutamate cysteine ligase catalytic subunit(GCLC)were significantly decreased(P<0.05).Compared with group T1,the ROS content,Ca^(2+)concentration and Keap1 mRNA relative expression level in GCs of group T2 were significantly decreased(P<0.05),and the mitochondrial membrane potential and mRNA relative expression levels of Nrf2,SOD1 and glutamate cysteine liga

关 键 词：蛋鸡 卵巢颗粒细胞 氧化应激 Nrf2-Keap1信号通路 黄豆苷元 藤茶黄酮 

分 类 号：S816.7[农业科学—饲料科学]