Effects of miRNA-206 on the Expression of Lipid Synthesis-related Genes and Fatty Acid Composition in Bovine (Bos taurus) Mammary Epithelial Cells
YAO Da-Wei1, WANG Tian-Zhen1,2, MA Jing1,2, YANG Chun-Lei1, CHEN Li-Li1, SUN Huan1,3, BAI Hai4, SONG Wen-Qin2, MA Yi1*
1. Tianjin Institute of Animal Husbandry and Veterinary Medicine,Tianjin 300381,China; 2. College of Life Sciences,Nankai University,Tianjin 300071,China; 3. College of Animal Science and Technology,Hebei Agricultural University,Baoding 071000,China; 4. College of Life Sciences,Shanxi Datong University,Datong 037009,China
Abstract：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 decreased intracellular triglyceride content and changed fatty acid composition,which suggested that miR-206 might play an important role in bovine fat metabolism. This study provided basic information for further study on the molecular mechanism of miR-206 in regulating lipid metabolism in bovine breast epithelial cells.
姚大为, 王添祯, 马静, 杨春蕾, 陈丽丽, 孙欢, 白海, 宋文芹, 马毅. 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[J]. 农业生物技术学报, 2020, 28(11): 1985-1993.
YAO Da-Wei, WANG Tian-Zhen, MA Jing, YANG Chun-Lei, CHEN Li-Li, SUN Huan, BAI Hai, SONG Wen-Qin, MA Yi. Effects of miRNA-206 on the Expression of Lipid Synthesis-related Genes and Fatty Acid Composition in Bovine (Bos taurus) Mammary Epithelial Cells. 农业生物技术学报, 2020, 28(11): 1985-1993.
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