豆蔻明通过激活芳香烃受体/核因子红系2相关因子2/NOD样受体热蛋白结构域相关蛋白3信号通路减轻脂多糖诱导的猪小肠上皮细胞的氧化应激和炎症反应  

Cardamonin Alleviates Oxidative Stress and Inflammatory Response of IPEC-J2 Cells Induced by Lipopolysaccharide by Activating Aromatic Hydrocarbon Receptor/Nuclear Factor Erythroid 2-Related Factor 2/Nod-Like Receptor Pyrin Domain-Associated Protein 3 Pathway

作　　者：方懿 张亚磊 符清瑶 邓谭杰 吴小妹 荀文娟 FANG Yi;ZHANG Yalei;FU Qingyao;DENG Tanjie;WU Xiaomei;XUN Wenjuan(College of Animal Science and Technology,Hainan University,Haikou 570228,China)

机构地区：[1]海南大学动物科技学院,海口570228

出　　处：《动物营养学报》2024年第2期1193-1203,共11页CHINESE JOURNAL OF ANIMAL NUTRITION

基　　金：国家自然科学基金地区基金项目(32060757)。

摘　　要：本试验旨在研究豆蔻明(CDN)在猪小肠上皮细胞(IPEC-J2)中通过芳香烃受体/核因子红系2相关因子2/NOD样受体热蛋白结构域相关蛋白3(AhR/Nrf2/NLRP3)信号通路对抗氧化应激和抗炎的作用机制。用分别含有0、1、3、10、30、60、100μmol/L CDN的培养液处理IPEC-J2细胞2 h,通过CCK-8法检测细胞活力,确定最佳浓度。将IPEC-J2细胞分为对照组(CON组)、CDN组、脂多糖组(LPS组)、LPS+CDN组(L+C组),每组3个重复。CON组细胞不做任何处理,LPS组添加1μg/mL LPS,L+C组添加1μg/mL LPS和最佳浓度CDN一同处理,之后收集细胞和上清。利用酶联免疫吸附试验(ELISA)检测细胞因子含量,实时荧光定量PCR检测相关基因mRNA相对表达量及试剂盒检测抗氧化能力。结果表明:当CDN浓度为60μmol/L时,IPEC-J2细胞活力与CON组无显著差异(P>0.05),表明该CDN浓度对IPEC-J2细胞是安全的,因此,后续选用60μmol/L CDN处理IPEC-J2细胞。与CON组相比,LPS组IPEC-J2细胞中白细胞介素-6(IL-6)、白细胞介素-1β(IL-1β)的含量显著升高(P<0.05),白细胞介素-10(IL-10)的含量显著减少(P<0.05);丙二醛(MDA)含量显著升高(P<0.05),谷胱甘肽(GSH)含量、谷胱甘肽过氧化物酶(GSH-Px)活性和总抗氧化能力(T-AOC)显著降低(P<0.05);Nrf2、醌氧化还原酶1(NQO1)和细胞色素P4501A1(CYP1A1)mRNA相对表达量显著降低(P<0.05),Kelch样ECH关联蛋白1(Keap1)、NLRP3、凋亡相关斑点样蛋白(ASC)与半胱天冬酶-1(Caspase-1)mRNA相对表达量显著升高(P<0.05);与LPS组相比,L+C组IPEC-J2细胞中IL-10的含量显著升高(P<0.05),IL-1β的含量显著减少(P<0.05);MDA含量显著降低(P<0.05),GSH含量以及SOD、CAT、GSH-Px活性和T-AOC显著升高(P<0.05);Nrf2、NQO1、血红素加氧酶-1(HO-1)及CYP1A1 mRNA相对表达量显著升高(P<0.05),Keap1、NLRP3、ASC与Caspase-1 mRNA相对表达量显著降低(P<0.05)。综上所述,CDN在IPEC-J2细胞中通过激活AhR/Nrf2/NLRP3信号通路提高抗氧化酶活性并启动下游This study was conducted to investigate the anti-oxidative stress and anti-inflammatory effects of cardamonin(CDN)in IPEC-J2 cells through the aromatic hydrocarbon receptor/nuclear factor erythroid 2-related factor 2/Nod-like receptor pyrin domain-associated protein 3(AhR/Nrf2/NLRP3)signaling pathway.IPEC-J2 cells were treated with 0,1,3,10,30,60 or 100μmol/mL CDN for 2 h,and cell viability was detected by CCK-8 assay to determine the optimal concentration.IPEC-J2 cells were divided into control group(CON group),CDN group,lipopolysaccharide group(LPS group)and LPS+CDN group(L+C group)with 3 replicates per group.The cells in the CON group were not treated,the LPS group was treated with 1μg/mL LPS,and the L+C group was treated with 1μg/mL LPS and the optimal concentration CDN,and then the cells and supernatant were collected.Cytokines contents were determined using ELISA kit,mRNA relative expression of related genes was detected using real-time fluorescent quantitative PCR,and the antioxidant capacity was detected by kit.The results showed as follows:when the concentration of CDN was 60μmol/mL,there was no significant difference in cell viability of IPEC-J2 compared with the CON group(P>0.05),indicating that the concentration of CDN selected in this experiment was safe for IPEC-J2 cells.Compared with CON group,the contents of interleukin-6(IL-6)and interleukin-1β(IL-1β)in IPEC-J2 cells were significantly increased(P<0.05),while the content of interleukin-10(IL-10)was significantly decreased(P<0.05);malondialdehyde(MDA)content was significantly increased(P<0.05),while glutathione(GSH)content,glutathione peroxidase(GSH-Px)activity and total antioxidant capacity(T-AOC)were significantly decreased(P<0.05);the relative expression levels of Nrf2,quinone oxidoreductase 1(NQO1)and cytochrome P4501A1(CYP1A1)mRNA were significantly decreased(P<0.05),and the relative expression levels of Kelch-like ECH-associated protein 1(Keap1),NLRP3,apoptosis-associated speck-like protein(ASC)and Caspase-1 mRNA were significantly inc

关 键 词：豆蔻明 脂多糖 IPEC-J2 氧化应激 

分 类 号：S816[农业科学—饲料科学]