Gansu Key Laboratory of Herbivorous Animal Biotechnology, Faculty of Animal Science and Technology, Gansu Agricultural University, Lanzhou 730070, China.
Gene-Marker Laboratory, Faculty of Agriculture and Life Science, Lincoln University, Lincoln 7647, New Zealand.
Food Funct. 2021 Oct 4;12(19):9432-9442. doi: 10.1039/d1fo01260f.
The microRNA (miR)-432 is differentially expressed in the mammary gland of two breeds of lactating sheep with different milk production traits, and between the non-lactating and peak-lactation periods, but there have been no reports describing the molecular mechanisms involved. In this study, the effect of miR-432 on the proliferation of ovine mammary epithelial cells (OMECs) and the target genes of miR-432 were investigated. The effects of miR-432 on the expression of the target genes and the content of triglycerides in the OMECs were also analyzed. Transfection with a miR-432 mimic was found using CCK8 and Edu assays, to inhibit the viability of OMECs and reduce the number of proliferated OMECs. In contrast, a miR-432 inhibitor had the opposite effect to the miR-432 mimic, and together these results suggest that miR-432 inhibits the proliferation of OMECs. A dual luciferase assay revealed that the genes for stearoyl-CoA desaturase () and lipoprotein lipase () are targeted by miR-432. The transfection of miR-432 mimic into OMECs resulted in decreases in the expression of and , and three other milk fat synthesis marker genes; , and . The mimic also decreased the content of triglycerides. The miR-432 inhibitor had the opposite effect to the mimic on the expression of these genes and the level of triglycerides. This is the first study to reveal the biological mechanisms by which miR-432 inhibits milk fat synthesis in sheep.
miR-432 在产奶量不同的两种泌乳绵羊的乳腺中以及非泌乳期和泌乳高峰期都有差异表达,但尚未有报道描述其涉及的分子机制。本研究旨在探讨 miR-432 对绵羊乳腺上皮细胞(OMEC)增殖的影响及其靶基因。还分析了 miR-432 对靶基因表达和 OMEC 中甘油三酯含量的影响。CCK8 和 Edu 检测发现,miR-432 模拟物转染可抑制 OMEC 的活力并减少增殖的 OMEC 数量。相反,miR-432 抑制剂的作用与 miR-432 模拟物相反,这些结果表明 miR-432 抑制 OMEC 的增殖。双荧光素酶报告基因检测表明,硬脂酰辅酶 A 去饱和酶()和脂蛋白脂肪酶()基因是 miR-432 的靶基因。miR-432 模拟物转染 OMEC 后,和三个其他乳脂肪合成标记基因、和的表达水平降低。该模拟物还降低了甘油三酯的含量。miR-432 抑制剂对这些基因和甘油三酯水平的表达产生了与模拟物相反的影响。这是首次揭示 miR-432 抑制绵羊乳脂肪合成的生物学机制的研究。