He Hongbing, Cai Mingcheng, Zhu Jiaoyue, Xiao Wudian, Liu Buwei, Shi Yu, Yang Xue, Liang Xiaohu, Zheng Tianhao, Hu Shenqiang, Jia Xianbo, Chen Shiyi, Wang Jie, Qin Yinghe, Lai Songjia
Farm Animal Genetic Resources Exploration and Innovation Key Laboratory of Sichuan Province, Sichuan Agricultural University, Sichuan, Chengdu, 611130, China.
College of Animal Science and Technology, China Agricultural University, Yuanmingyuan West Road, Haidian District, 100193, Beijing, People's Republic of China.
In Vitro Cell Dev Biol Anim. 2018 Mar;54(3):241-249. doi: 10.1007/s11626-018-0232-z. Epub 2018 Feb 9.
Although emerging data support crucial roles for microRNAs (miRNAs) during adipogenesis, the detailed mechanisms remain largely unknown. In this study, it was shown that in rabbits, levels of miR-148a-3p not only increased in white adipose tissue during early stages of growth but also during in vitro cultured preadipocyte differentiation. Furthermore, overexpression of miR-148a-3p significantly upregulated the mRNA levels of PPARγ, C/EBPα, and FABP4, as well as the protein levels of PPARγ, as indicated by qPCR and western blotting analyses. Overexpression of miR-148a-3p also promoted intracellular triglyceride accumulation. In contrast, downregulation of miR-148a-3p inhibited the differentiation of rabbit preadipocytes. Next, based on target gene prediction and a luciferase reporter assay, we further demonstrated that miR-148a-3p directly targeted one of the 3' untranslated regions of PTEN. Finally, it was observed inhibition of PTEN by siRNA promoted rabbit preadipocyte differentiation. Taken together, our results suggested that miR-148a-3p could be involved in regulating rabbit preadipocyte differentiation through inhibiting expression of PTEN, which further highlighted the importance of miRNAs during adipogenesis.
尽管新出现的数据支持微小RNA(miRNA)在脂肪生成过程中发挥关键作用,但详细机制仍 largely 未知。在本研究中,结果表明,在兔子中,miR-148a-3p的水平不仅在生长早期的白色脂肪组织中升高,而且在体外培养的前脂肪细胞分化过程中也升高。此外,如qPCR和蛋白质印迹分析所示,miR-148a-3p的过表达显著上调了PPARγ、C/EBPα和FABP4的mRNA水平,以及PPARγ的蛋白质水平。miR-148a-3p的过表达还促进了细胞内甘油三酯的积累。相反,miR-148a-3p的下调抑制了兔子前脂肪细胞的分化。接下来,基于靶基因预测和荧光素酶报告基因检测,我们进一步证明miR-148a-3p直接靶向PTEN的3'非翻译区之一。最后,观察到通过siRNA抑制PTEN可促进兔子前脂肪细胞分化。综上所述,我们的结果表明miR-148a-3p可能通过抑制PTEN的表达参与调节兔子前脂肪细胞分化,这进一步突出了miRNA在脂肪生成过程中的重要性。