College of Animal Science and Veterinary Medicine, Heilongjiang Bayi Agricultural University, Daqing, China.
Agricultural College, Guangdong Ocean University, Zhanjiang, China.
J Anim Physiol Anim Nutr (Berl). 2019 Sep;103(5):1365-1373. doi: 10.1111/jpn.13162. Epub 2019 Jul 29.
According to our previous studies, bta-miR-152, PRKAA1 and UCP3 are differentially expressed in mammary gland tissues of high milk fat and low milk fat cows, and the trend in bta-miR-152 expression is opposite from those of PRKAA1 and UCP3. To further identify the function and regulatory mechanism of bta-miR-152 in milk fat metabolism, we investigated the effect of bta-miR-152 on cellular triglyceride content in bovine mammary epithelial cells cultured in vitro, on the basis of bta-miR-152 overexpression and inhibition assays. The target genes of bta-miR-152 were identified through qPCR, Western blotting and dual luciferase reporter gene detection. Compared with that in the control group, the expression of UCP3 was significantly lower in the bta-miR-152 mimic group, the expression of PRKAA1 was decreased, and the intracellular TAG content was significantly increased. After transfection with bta-miR-152 inhibitor, the expression of UCP3 increased significantly, and the expression of PRKAA1 decreased, but the difference was not significant; in addition, the intracellular TAG content decreased significantly. Therefore, we concluded that bta-miR-152 affects the intracellular TAG content by targeting UCP3.
根据我们之前的研究,bta-miR-152、PRKAA1 和 UCP3 在高脂肪奶和低脂肪奶奶牛的乳腺组织中表达存在差异,bta-miR-152 的表达趋势与 PRKAA1 和 UCP3 相反。为了进一步确定 bta-miR-152 在乳脂代谢中的功能和调节机制,我们基于 bta-miR-152 的过表达和抑制实验,研究了 bta-miR-152 对体外培养的牛乳腺上皮细胞中细胞内甘油三酯含量的影响。通过 qPCR、Western blot 和双荧光素酶报告基因检测鉴定了 bta-miR-152 的靶基因。与对照组相比,bta-miR-152 模拟物组 UCP3 的表达显著降低,PRKAA1 的表达降低,细胞内 TAG 含量显著增加。转染 bta-miR-152 抑制剂后,UCP3 的表达显著增加,PRKAA1 的表达降低,但差异不显著;此外,细胞内 TAG 含量显著降低。因此,我们得出结论,bta-miR-152 通过靶向 UCP3 影响细胞内 TAG 含量。