MicroRNA-432 inhibits milk fat synthesis by targeting SCD and LPL in ovine mammary epithelial cells
MicroRNA-432 inhibits milk fat synthesis by targeting SCD and LPL in ovine mammary epithelial cells
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MicroRNA-432 通过靶向绵羊乳腺上皮细胞中的 SCD 和 LPL 抑制乳脂合成
DOI：
10.1039/d1fo01260f
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发表时间：
2021-08-16
期刊：
影响因子：
6.1
通讯作者：
Huang, Zhaochun
中科院分区：
文献类型：
--
作者：
Hao, Zhiyun;Luo, Yuzhu;Huang, Zhaochun
微小RNA（miR）-432在具有不同产奶性状的两个品种的泌乳绵羊乳腺中以及在非泌乳期和泌乳高峰期之间差异表达，但尚未有报道描述所涉及的分子机制。在本研究中，研究了miR - 432对绵羊乳腺上皮细胞（OMECs）增殖的影响以及miR - 432的靶基因。还分析了miR - 432对靶基因表达和OMECs中甘油三酯含量的影响。通过CCK8和Edu检测发现，用miR - 432模拟物转染可抑制OMECs的活力并减少增殖的OMECs数量。相反，miR - 432抑制剂与miR - 432模拟物的作用相反，这些结果共同表明miR - 432抑制OMECs的增殖。双荧光素酶检测显示，硬脂酰 - 辅酶A去饱和酶（SCD）和脂蛋白脂肪酶（LPL）的基因是miR - 432的靶基因。将miR - 432模拟物转染到OMECs中导致SCD和LPL以及其他三个乳脂合成标记基因FABP4、LPIN1和ACACA的表达降低。该模拟物还降低了甘油三酯含量。miR - 432抑制剂对这些基因的表达和甘油三酯水平的作用与模拟物相反。这是第一项揭示miR - 432抑制绵羊乳脂合成的生物学机制的研究。
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 (SCD) and lipoprotein lipase (LPL) are targeted by miR-432. The transfection of miR-432 mimic into OMECs resulted in decreases in the expression of SCD and LPL, and three other milk fat synthesis marker genes; FABP4, LPIN1 and ACACA. 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.