Jin Xiayang, Hao Zhiyun, Zhao Mengli, Shen Jiyuan, Ke Na, Song Yize, Qiao Lirong, Lu Yujie, Hu Liyan, Wu Xinmiao, Wang Jiqing, Luo Yuzhu
Gansu Key Laboratory of Herbivorous Animal Biotechnology, Faculty of Animal Science and Technology, Gansu Agricultural University, Lanzhou 730070, China.
Animals (Basel). 2021 Mar 15;11(3):820. doi: 10.3390/ani11030820.
MicroRNAs (miRNAs) have been found to be involved in lipid deposition and metabolism. However, there have been no reports on the roles of miR-148a in the proliferation and adipogenesis of preadipocytes in sheep. In this study, the expression of miR-148a was profiled in the eight tissues of Tibetan ewes and differentiated preadipocytes, and the role of miR-148a in differentiation and proliferation of ovine preadipocytes was investigated using Oil Red O staining, CCK-8, EdU staining, cell cycle detection, and RT-qPCR. The effect of on the differentiation of ovine preadipocytes was also investigated. The miR-148a was widely expressed in the eight tissues investigated and had significantly increased expression in liver, spleen and subcutaneous adipose tissues, and the heart. The expression of miR-148a continued to increase with the differentiation of ovine preadipocytes. The over-expression of miR-148a significantly promoted differentiation but inhibited the proliferation of ovine preadipocytes. The inhibition of miR-148a had the opposite effect on the differentiation and proliferation of ovine preadipocytes with over-expressed miR-148a. The results from the dual luciferase reporter assays showed that miR-148a mimic significantly decreased the luciferase activity of -3'UTR dual luciferase reporter vector, suggesting that is a target gene of miR-148a. In over-expressed- preadipocytes, the number of lipid droplets remarkably decreased, and the expression levels of adipogenesis marker genes , , , , and were also significantly down-regulated. These results suggest that miR-148a accelerated the adipogenic differentiation of ovine preadipocytes by inhibiting expression, and also inhibited the proliferation of ovine preadipocytes.
微小RNA（miRNA）已被发现参与脂质沉积和代谢。然而，关于miR-148a在绵羊前体脂肪细胞增殖和脂肪生成中的作用尚无报道。在本研究中，检测了miR-148a在藏羊八个组织和分化的前体脂肪细胞中的表达情况，并采用油红O染色、CCK-8法、EdU染色、细胞周期检测和RT-qPCR技术研究了miR-148a在绵羊前体脂肪细胞分化和增殖中的作用。还研究了[此处原文缺失相关内容]对绵羊前体脂肪细胞分化的影响。miR-148a在所检测的八个组织中广泛表达，在肝脏、脾脏、皮下脂肪组织和心脏中表达显著增加。随着绵羊前体脂肪细胞的分化，miR-148a的表达持续增加。miR-148a的过表达显著促进了绵羊前体脂肪细胞的分化，但抑制了其增殖。抑制miR-148a对过表达miR-148a的绵羊前体脂肪细胞的分化和增殖产生相反的影响。双荧光素酶报告基因检测结果表明，miR-148a模拟物显著降低了[此处原文缺失相关内容]-3'UTR双荧光素酶报告基因载体的荧光素酶活性，表明[此处原文缺失相关内容]是miR-148a的靶基因。在过表达[此处原文缺失相关内容]的前体脂肪细胞中，脂滴数量显著减少，脂肪生成标记基因[此处原文缺失相关内容]、[此处原文缺失相关内容]、[此处原文缺失相关内容]、[此处原文缺失相关内容]、[此处原文缺失相关内容]和[此处原文缺失相关内容]的表达水平也显著下调。这些结果表明，miR-148a通过抑制[此处原文缺失相关内容]的表达加速了绵羊前体脂肪细胞的成脂分化，同时也抑制了绵羊前体脂肪细胞的增殖。
