bta-miR-29d-3p对奶牛乳腺上皮细胞乳脂代谢相关基因表达及甘油三酯含量的影响  

Effect of bta-miR-29d-3p on the expression of genes related to milk lipid metabolism and triglyceride content in dairy cows mammary epithelial cells

作　　者：赵淑颖 姚大为[2,3,4] 赵欣 杨春蕾 曹彦怡[6] 张漫 丁向彬 马毅 ZHAO Shuying;YAO Dawei;ZHAO Xin;YANG Chunlei;CAO Yanyi;ZHANG Man;DING Xiangbin;MA Yi(College of Animal Science and Animal Medicine,Tianjin Agricultural University,Xiqing,Tianjin 300384,China;Institute of Animal Science and Veterinary,Tianjin Academy of Agricultural Sciences,Xiqing,Tianjin 300381,China;Tianjin Key Laboratory of Animal Molecular Breeding and Biotechnology,Xiqing,Tianjin 300381,China;Tianjin Engineering Research Center of Animal Healthy Farming,Xiqing,Tianjin 300381,China;College of Life Science,Nankai University,Nankai,Tianjin 300071,China;School of Integrative Medicine,Tianjin University of Traditional Chinese Medicine,Jinghai,Tianjin 301600,China;Institute of Agricultural Quality Standards and Testing Technology,Tianjin Academy of Agricultural Sciences,Xiqing,Tianjin 300381,China)

机构地区：[1]天津农学院动物科学与动物医学学院,天津西青300384 [2]天津市农业科学院畜牧兽医研究所,天津西青300381 [3]天津市畜禽分子育种与生物技术重点实验室,天津西青300381 [4]天津市畜禽健康养殖工程技术中心,天津西青300381 [5]南开大学生命科学学院,天津南开300071 [6]天津中医药大学中西医结合学院,天津静海301600 [7]天津市农业科学院农业质量标准与检测技术研究所,天津西青300381

出　　处：《中国饲料》2023年第11期13-19,共7页China Feed

基　　金：天津市农业科学院青年科研人员创新研究与实验项目(2021010);国家自然科学基金(31702095);天津市自然科学基金(18JCYBJC30200);天津市科技计划项目(19ZXZYSN00020,19ZXZYSN00130);种业创新研究(2022ZYCX010);2021年古浪县东西部协作科技创新项目-高产奶牛养殖技术与集成示范(2021-ZMZX-059)。

摘　　要：为探索bta-miR-29d-3p与奶牛乳腺上皮细胞乳脂代谢相关基因表达及甘油三酯含量之间的调控关系,试验设计并合成bta-miR-29d-3p的模拟物和抑制物,通过瞬时转染技术将bta-miR-29d-3p的模拟物和抑制物转染到奶牛乳腺上皮细胞中,利用实时荧光定量PCR(RT-qPCR)技术检测bta-miR-29d-3p抑制或过表达后奶牛乳腺上皮细胞乳脂合成相关基因表达水平的变化,细胞内甘油三酯含量用试剂盒检测。结果显示:bta-miR-29d-3p过表达后甘油三酯合成相关基因二酰甘油酰基转移酶(DGAT1)、线粒体甘油-3-磷酸酰基转移酶(GPAM),脂肪酸合成相关基因乙酰辅酶A羧化酶A(ACACA)、脂肪酸合成酶(FASN),转录因子固醇调节元件结合蛋白1(SREBF1)的表达量显著降低(P<0.01),肝X受体Α(LXRA)表达量显著上升(P<0.01)。同时过表达bta-miR-29d-3p奶牛乳腺上皮细胞中甘油三酯含量显著降低(P<0.05)。干扰bta-miR-29d-3p显著增加了DGAT1、ACACA、FASN、GPAM、转录因子过氧化物酶体增殖物激活受体γ(PPARG1)(P<0.01)和SREBF1(P<0.05)的表达量,同时甘油三酯含量显著增加(P<0.05)。说明bta-miR-29d-3p通过改变乳脂代谢相关基因和甘油三酯水平,从而调节奶牛乳脂代谢。In order to explore the regulatory relationship between bta-miR-29d-3p and the expression of genes related to milk fat metabolism and triglyceride content in dairy cow mammary epithelial cells,the mimics and inhibitors of bta-miR-29d-3p were designed and synthesized.The simulants and repressor of bta-miR-29d-3p were transfected into cow mammary epithelial cells by transient transfection technology.The expression levels of milk fat synthesis related genes in cow mammary epithelial cells after bta-miR-29d-3p inhibition or overexpression were detected by real-time fluorescent quantitative PCR(RT-qPCR).The intracellular triglyceride content was detected by kit.The results showed that after overexpression of bta-miR-29d-3p,triglyceride synthesis-related genes diacylglycerol acyltransferase(DGAT1),mitochondrial glycerol 3-phosphate acyltransferase(GPAM),fatty acid synthesis-related genes acetyl-CoA carboxylase A(ACACA)and fatty acid synthetase(FASN),the expression of sterol regulatory element binding protein 1(SREBF1)was significantly decreased(P<0.01),the expression level of liver X receptorα(LXRA)increased significantly(P<0.01).At the same time,the triglyceride content in mammary epithelial cells of cows overexpressing bta-miR-29d-3p significantly decreased(P<0.05).Interference with bta-miR-29d-3p significantly increased DGAT1,ACACA,FASN,GPAM,peroxisome proliferator-activated receptorγ-1(PPARG1)(P<0.01)and SREBF1(P<0.05),and triglyceride content was significantly increased(P<0.05).These results indicated that bta-miR-29d-3p regulates milk fat metabolism in dairy cows by changing the genes related to milk fat metabolism and triglyceride levels.

关 键 词：bta-miR-29d-3p 乳脂代谢 甘油三酯 奶牛乳腺上皮细胞 

分 类 号：S811[农业科学—畜牧学]