Yang W C, Guo W L, Zan L S, Wang Y N, Tang K Q
J Anim Sci. 2017 Jul;95(7):2898-2906. doi: 10.2527/jas.2017.1504.
Milk fat determines the quality of milk and is also a main targeted trait in dairy cow breeding. Recent studies have revealed important regulatory roles of microRNAs (miRNA) in milk fat synthesis in the mammary gland. However, the role of miRNA in bovine mammary epithelial cells (BMEC) remains largely unknown. In this study, we found that the overexpression of miR-130a significantly decreased cellular triacylglycerol (TAG) levels and suppressed lipid droplet formation, whereas the inhibition of miR-130a resulted in greater lipid droplet formation and TAG accumulation in BMEC. MiR-130a also significantly affected mRNA expression related to milk fat metabolism. Specifically, the overexpression of miR-130a reduced the mRNA expression of , , , and , whereas the downregulation of miR-130a increased the mRNA expression of , , , , , and . Furthermore, western blot analysis revealed the protein level of PPARG in miR-130a mimic and inhibitor transfection groups to be consistent with the mRNA expression response. Finally, luciferase reporter assays verified that PPARG was the direct target of miR-130a. This study provides the first experimental evidence that miR-130a directly affects TAG synthesis in BMEC by targeting PPARG, suggesting that miR-130a potentially could be used to improve beneficial milk components in dairy cows.
乳脂肪决定了牛奶的品质,也是奶牛育种中的一个主要目标性状。最近的研究揭示了微小RNA(miRNA)在乳腺乳脂肪合成中的重要调控作用。然而,miRNA在牛乳腺上皮细胞(BMEC)中的作用仍 largely未知。在本研究中,我们发现miR-130a的过表达显著降低了细胞三酰甘油(TAG)水平并抑制了脂滴形成,而抑制miR-130a则导致BMEC中脂滴形成增加和TAG积累。miR-130a还显著影响与乳脂肪代谢相关的mRNA表达。具体而言,miR-130a的过表达降低了 、 、 、 和 的mRNA表达,而miR-130a的下调增加了 、 、 、 、 和 的mRNA表达。此外,蛋白质印迹分析显示,在miR-130a模拟物和抑制剂转染组中PPARG的蛋白质水平与mRNA表达反应一致。最后,荧光素酶报告基因测定证实PPARG是miR-130a的直接靶标。本研究提供了首个实验证据,即miR-130a通过靶向PPARG直接影响BMEC中的TAG合成,表明miR-130a可能可用于改善奶牛的有益乳成分。