miRNA-206对牛乳腺上皮细胞脂质合成相关基因表达及脂肪酸组成的影响  

Effects of miRNA-206 on the Expression of Lipid Synthesis-related Genes and Fatty Acid Composition in Bovine(Bos taurus)Mammary Epithelial Cells

作　　者：姚大为[1] 王添祯 马静 杨春蕾[1] 陈丽丽 孙欢 白海 宋文芹[2] 马毅[1] YAO Da-Wei;WANG Tian-Zhen;MA Jing;YANG Chun-Lei;CHEN Li-Li;SUN Huan;BAI Hai;SONG Wen-Qin;MA Yi(Tianjin Institute of Animal Husbandry and Veterinary Medicine,Tianjin 300381,China;College of Life Sciences,Nankai University,Tianjin 300071.China;College of Animal Science and Technology,Hebei Agricultural University,Baoding 071000,China;College of Life Sciences,Shanxi Datong University,Datong 037009,China)

机构地区：[1]天津市畜牧兽医研究所,天津300381 [2]南开大学生命科学学院,天津300071 [3]河北农业大学动物科技学院,保定071000 [4]山西大同大学生命科学学院,大同037009

出　　处：《农业生物技术学报》2020年第11期1985-1993,共9页Journal of Agricultural Biotechnology

基　　金：天津市自然科学基金(18JCYBJC30200);国家自然科学基金(31702095)。

摘　　要：牛奶中的脂肪酸含量及成分决定着牛奶的品质,而miRNA能够调控哺乳动物乳腺中脂质代谢途径,进而影响脂质含量及其成分。本研究旨在通过miRNA转染技术揭示miR-206对牛(Bos taurus)乳腺上皮细胞(bovine mammary epithelial cells,BMECs)中脂质合成相关基因表达、甘油三酯含量及脂肪酸组成的影响。在牛乳腺上皮细胞中转染60 nmol/L miR-206模拟物和抑制物,应用qRT-PCR技术和生化方法分别检测脂质合成相关基因的表达和细胞内甘油三酯含量,使用气相色谱检测脂肪酸组成的变化。结果表明,miR-206模拟物抑制甘油三酯合成相关基因线粒体甘油-3-磷酸酰基转移酶(glycerol-3-phosphate acyltransferase mitochondrial,GPAM)、磷酸甘油酰基转移酶6(1-acylglycerol-3-phosphate O-acyltransferase6,AGPAT6)、二酰甘油酰基转移酶1(diacylglycerol O-acyltransferase 1,DGAT1)、脂素1(lipin 1,LPIN1)的表达(P<0.01),细胞内甘油三酯含量显著减少(P<0.05);固醇调节元件结合蛋白1(sterol-regulatory element binding proteins 1,SREBP1)、脂肪酸合成酶(fatty acid synthase,FASN)、乙酰辅酶A羧化酶α(acetyl-CoA carboxylaseα,ACACA)和长链脂肪酸延长酶6(long chain fatty acid elongase 6,ELOVL6)的表达量也有所下调,脂肪酸成分发生改变,其中C14∶0(P<0.01)和C22∶6n3(P<0.05)含量显著降低,C16∶1含量显著增加(P<0.01)。miR-206抑制物则促进GPAM、AGPAT6、DGAT1和LPIN1表达(P<0.01),细胞内甘油三酯含量增加,SREBP1、FASN、ACACA和ELOVL6表达量显著上调(P<0.01),但脂肪酸成分未发生改变。综上所述,miR-206对牛乳腺上皮细胞脂质合成相关基因的表达具有负调控作用,导致细胞内甘油三酯含量减少以及脂肪酸组成改变,提示miR-206在牛乳腺脂代谢中可能具有重要作用,本研究为深入探讨miR-206调控牛乳腺上皮细胞脂质代谢的分子机理提供基础资料。The content and components of fatty acids in milk determine the quality of milk,while miRNAs can regulate the lipid metabolism pathway in mammary glands of mammals,thereby affect lipid content and components.The purpose of this study is to reveal the effects of miR-206 on the expression of genes related to lipid synthesis,triglyceride content and fatty acid composition in bovine(Bos taurus)mammary epithelial cells(BMECs)by miRNA transfection.In this study,60 nmol/L miR-206 mimics and inhibitor were transfected into bovine mammary epithelial cells,then qRT-PCR and biochemical experiment were used to detect the mRNA level of lipid synthesis-related genes,intracellular triglyceride content,respectively.and the changes of fatty acid composition were detected by gas chromatography.The results showed that when transfected with miR-206 mimics,the mRNA level of triglyceride synthesis-related genes including mitochondrial glycerol-3-phosphate acyltransferase(GPAM),1-acylglycerol-3-phosphate O-acyltransferase 6(AGPAT6),diacylglycerol O-acyltransferase 1(DGAT1)and lipin 1(LPIN1)were significantly down-regulated(P<0.01)with significant reduction of intracellular triglyceride content(P<0.05).The expression of sterol-regulatory element binding proteins 1(SREBP1),fatty acid synthase(FASN),acetyl-CoA carboxylase α(ACACA)and long chain fatty acid elongase 6(ELOVL6)were down-regulated and fatty acid composition changed,in which the content of C14∶0(P<0.01)and C22∶6n3(P<0.05)were significantly reduced,and the content of C16∶1(P<0.01)was significantly increased.When transfected with miR-206 inhibitor in bovine mammary epithelial cells,the expression of GPAM,AGPAT6,DGAT1 and LPIN1 were significantly up-regulated(P<0.01)and the intracellular triglyceride content increased,combined with up-regulated SREBP1,FASN,ACACA and ELOVL6(P<0.01),but the fatty acid composition did not change.In summary,miR-206 had a negative regulatory effect on the expression of lipid synthesis-related genes in bovine mammary epithelial cells,also led to

关 键 词：牛乳腺上皮细胞(BMEC) miR-206 脂代谢 

分 类 号：S287[农业科学—农业水土工程] S813.3[农业科学—农业工程]