2,2’-偶氮二(2-脒基丙烷)二盐酸盐诱导的氧化应激对猪肠上皮细胞活力及线粒体功能的影响  

Effects of 2,2’⁃Azobis(2⁃Amidinopropane)Dihydrochloride⁃Induced Oxidative Stress on Cell Viability and Mitochondrial Function of Porcine Intestinal Epithelial Cells

作　　者：鲁慧杰 张莹 田志梅 容庭 刘志昌 邓盾 马现永 LU Huijie;ZHANG Ying;TIAN Zhimei;RONG Ting;LIU Zhichang;DENG Dun;MA Xianyong(State Key Laboratory of Livestock and Poultry Breeding,Key Laboratory of Animal Nutrition and Feed Science in South China,Ministry of Agriculture and Rural Affairs,Guangdong Key Laboratory of Animal Breeding and Nutrition,Guangdong Engineering Technology Research Center of Animal Meat Quality and Safety Control and Evaluation,Institute of Animal Science,Guangdong Academy of Agricultural Sciences,Guangzhou 510640,China;Maoming Branch,Guangdong Laboratory for Lingnan Modern Agriculture,Maoming 525000,China)

机构地区：[1]广东省农业科学院动物科学研究所,畜禽育种国家重点实验室,农业农村部华南动物营养与饲料重点实验室,广东省畜禽育种与营养研究重点实验室,广东畜禽肉品质量安全控制与评定工程技术研究中心,广州510640 [2]岭南现代农业科学与技术广东省实验室茂名分中心,茂名525000

出　　处：《动物营养学报》2021年第7期4038-4048,共11页CHINESE JOURNAL OF ANIMAL NUTRITION

基　　金：广东省基础与应用基础研究基金项目(2020A1515010591);广州市科技计划项目(202102020549);广东省农业科学院人才培养计划(R2017YJ⁃YB3007,R2018QD⁃0718,R2017PY⁃JG001);广东省现代农业产业技术体系饲料产业创新团队(2020KJ115)。

摘　　要：本试验旨在评估2,2’-偶氮二(2-脒基丙烷)二盐酸盐(AAPH)诱导的氧化应激对猪肠上皮细胞(IPEC⁃J2细胞)活力及线粒体功能的影响,以期建立一种稳定的体外研究肠道氧化应激的模型。以不同浓度(0、30、32、34 mmol/L)AAPH处理IPEC⁃J2细胞24 h后,检测细胞活力、细胞增殖、细胞内总活性氧(ROS)含量、线粒体膜电位、线粒体DNA(mtDNA)拷贝数以及线粒体功能相关基因[线粒体转录因子A(TFAM)、线粒体转录因子B1(TFB1M)、线粒体转录因子B2(TFB2M)、解耦连蛋白1(UCP1)、解耦连蛋白2(UCP2)、解耦连蛋白3(UCP3)]表达的变化。结果表明:AAPH对IPEC⁃J2细胞活力的影响呈剂量及作用时间依赖关系。与0 mmol/L AAPH处理相比,不同浓度(30、32、34 mmol/L)AAPH处理均显著抑制了IPEC⁃J2细胞增殖(P<0.05),细胞活力显著下降(P<0.05),细胞内总ROS含量显著升高(P<0.05),细胞线粒体功能相关基因(TFAM、TFB1M、TFB2M、UCP1、UCP3)表达显著下调(P<0.05),mtDNA拷贝数显著降低(P<0.05)。综上所述,AAPH诱导的氧化应激通过增加IPEC⁃J2细胞内总ROS含量引起细胞线粒体功能损伤,进而导致细胞活力下降,抑制细胞增殖。The aim of this study was to evaluate the effects of 2,2’⁃azobis(2⁃amidinopropane)dihydrochlo⁃ride(AAPH)⁃induced oxidative stress on cell viability and mitochondrial function of porcine intestinal epitheli⁃al cells(IPEC⁃J2 cells),and to establish a stable in vitro oxidative damage model.IPEC⁃J2 cells were treated with different concentrations(0,30,32 and 34 mmol/L)of AAPH for 24 h,the cell viability,cell prolifera⁃tion,intracellular total reactive oxygen species(ROS)content,mitochondria membrane potential,mitochon⁃drial DNA(mtDNA)copy number and the expression of mitochondrial function⁃related genes[mitochondrial transcription factor A(TFAM),mitochondrial transcription factor B1(TFB1M),mitochondrial transcription factor B2(TFB2M),uncoupling protein 1(UCP1),uncoupling protein 2(UCP2)and uncoupling protein 3(UCP3)]were measured.The results showed that AAPH had a dosage and time dependent relationship on the viability of IPEC⁃J2 cells.Compared with the 0 mmol/L AAPH treatment,different concentrations(30,32 and 34 mmol/L)of AAPH treatment significantly inhibited the proliferation of IPEC⁃J2 cells(P<0.05),the cell viability significantly decreased(P<0.05),the intracellular total ROS content significantly increased(P<0.05),the mRNA expression of mitochondrial function⁃related genes(TFAM,TFB1M,TFB2M,UCP1 and UCP3)significantly down⁃regulated(P<0.05),and the copy number of mtDNA significantly decreased(P<0.05).In summary,AAPH⁃induced oxidative damage leads to cell mitochondria dysfunction,resulting in de⁃crease of cell viability and inhibited cell proliferation of IPEC⁃J2 cells.

关 键 词：AAPH 氧化应激 肠上皮细胞 线粒体 

分 类 号：S811.2[农业科学—畜牧学]