奶山羊FTO基因对乳腺上皮细胞脂代谢相关基因表达及甘油三酯合成的影响  

Effects of FTO on the Expression of Lipid Metabolism-related Genes and Triglyceride Synthesis in Dairy Goat Mammary Epithelial Cells

作　　者：姚大为[1] 郭晓飞 刘玉 李玉鹏[1] 陈龙宾[1] 张金龙[1] 张效生[1] YAO Dawei;GUO Xiaofei;LIU Yu;LI Yupeng;CHEN Longbin;ZHANG Jinlong;ZHANG Xiaosheng(Institute of Animal Science and Veterinary,Tianjin Academy of Agricultural Sciences/Tianjin Key Laboratory of Animal Molecular Breeding and Biotechnology/Tianjin Engineering Research Center of Animal Healthy Farming,Tianjin 300381,China)

机构地区：[1]天津市农业科学院畜牧兽医研究所/天津市畜禽分子育种与生物技术重点实验室/天津市畜禽健康养殖工程技术中心,天津300381

出　　处：《家畜生态学报》2025年第1期17-25,共9页Journal of Domestic Animal Ecology

基　　金：天津市科技计划项目(24YDTPJC00480,24YDTPJC00290);畜禽生物育种全国重点实验室开放课题(2024SKLAB6-4,2024SKLAB6-11);甘肃省科技计划项目(24CXND004);天津市自然科学基金(23JCYBJC00260);天津市重点研发计划(23YFZCSN00170);天津市农业科学院财政创新种业研究项目(2024ZYCX012)。

摘　　要：试验旨在通过过表达和干扰技术探究脂肪与肥胖相关基因(fat mass and obesity associated,FTO)对奶山羊乳腺上皮细胞脂代谢相关基因的表达及甘油三酯合成的影响。通过克隆得到奶山羊(Capra hircus)乳腺组织FTO基因的CDS区序列,构建FTO过表达载体,并设计合成靶向FTO的siRNA(small interference RNA),利用实时荧光定量PCR(RT-qPCR)技术检测过表达或干扰FTO基因后奶山羊乳腺上皮细胞中脂代谢相关基因及甘油三酯含量的变化。结果表明,奶山羊FTO基因的CDS区全长1518 bp,与绵羊(Ovis aries)(NM_001104931.1)、牛(Bos taurus)(NM_001098142.1)和水牛(Bubalus bubalis)(XM_006043050.4)的序列相似性分别为99%、97%和96%。在乳腺上皮细胞中过表达FTO能够显著上调固醇调节元件结合蛋白1(sterol-regulatory element binding proteins 1,SREBF1)、脂肪酸合成酶(fatty acid synthase,FASN)、二酰甘油酰基转移酶2(diacylglycerol O-acyltransferase 2,DGAT2)的mRNA水平(P<0.05),显著下调甘油三酯水解相关基因如激素敏感脂酶(hormone-sensitive lipase,HSL)的表达水平(P<0.05),同时细胞中甘油三酯含量显著增加。在奶山羊乳腺上皮细胞中干扰FTO得到与之相反的结果。综上,FTO基因在调节奶山羊乳腺上皮细胞脂代谢中发挥重要的作用。This study aims to investigate the effects of the fat mass and obesity-associated gene(FTO)on the expression of lipid metabolism-related genes and triglyceride(TAG)synthesis in mammary epithelial cells(DMECs)of dairy goats through overexpression and interference techniques.The complete coding sequence(CDS)of the FTO gene in the mammary tissue of diary goats(Capra hircus)was cloned,and an FTO overexpression vector was constructed.Additionally,small interference RNA(siRNA)targeting FTO were designed and synthesized.RT-qPCR was used to detect changes in the expression of lipid metabolism-related genes and triglyceride content in DMECs after overexpressing or interfering with the FTO gene.The results indicated that the full length of the FTO gene CDS region in dairy goat was 1518 bp,with sequence homology of 99%,97% and 96% to sheep(Ovis aries)(NM_001104931.1),cattle(Bos taurus)(NM_001098142.1)and buffalo(Bubalus bubalis)(XM_006043050.4),respectively.Overexpression of FTO in DMECs significantly upregulated the mRNA levels of sterol-regulatory element binding protein 1(SREBF1),fatty acid synthase(FASN),and diacylglycerol O-acyltransferase 2(DGAT2)(P<0.05),while significantly downregulating the expression levels of triglyceride hydrolysis-related genes such as hormone-sensitive lipase(HSL)(P<0.05).Furthermore,the TAG content in the cells significantly increased.Conversely,interference with FTO in DMECs yielded opposite results.In conclusion,FTO gene plays an important role in regulating lipid metabolism in DMECs of dairy goats,providing a theoretical basis for improving the regulatory network of lipid metabolism in mammary gland of dairy goat.

关 键 词：奶山羊 乳腺上皮细胞 FTO 脂代谢 甘油三酯 

分 类 号：S827[农业科学—畜牧学]