Bai Jinhui, Xu Hongyan, Fang Jiachen, Zhang Congcong, Song Jixuan, Zhang Xinxin, Hao Beibei, Yin Baozhen, Xia Guangjun
Agriculture College, Yanbian University, Yanji, China.
Engineering Research Center of North-East Cold Region Beef Cattle Science & Technology Innovation, Ministry of Education, Yanbian University, Yanji, China.
Anim Biotechnol. 2023 Dec;34(7):2343-2352. doi: 10.1080/10495398.2022.2088552. Epub 2022 Jun 22.
MicroRNAs (miRNAs) are small, single-stranded, noncoding RNAs of approximately 21 to 23 nucleotides in length. Owing to their regulation of gene expression and many physiological processes including fat metabolism, they have become a popular research topic in recent years; however, the exact functional mechanisms by which they regulate fat metabolism have not been fully elucidated. Here, we identified miR-15a, which specifically acquired the 3' untranslated region (UTR) containing 4-aminobutyrate aminotransferase (ABAT), and validated the regulation of its expression and involvement in adipogenesis mechanisms. We used a dual-luciferase reporter assay and transfection-mediated miR-15a overexpression and inhibition in Yanbian yellow cattle preadipocytes to investigate the role of miR-15a in adipogenesis. The results showed that miR-15a directly targets the 3'UTR of ABAT and downregulates its expression. Additionally, at the protein and mRNA levels, miR-15a overexpression using a miRNA mimic inhibited triglyceride accumulation and downregulated lipogenic peroxisome proliferator-activated receptor γ and CCAAT enhancer-binding protein α, whereas miR-15a inhibition had the opposite effect. The above results indicated that miR-15a regulated the differentiation of Yanbian yellow cattle preadipocytes by inhibiting the expression of ABAT. Furthermore, our findings suggested that miR-15a and its target gene(s) might represent new targets for investigating intramuscular fat deposits in cattle and treating human obesity.
微小RNA(miRNA)是长度约为21至23个核苷酸的小单链非编码RNA。由于它们对基因表达以及包括脂肪代谢在内的许多生理过程具有调控作用,近年来已成为热门研究课题;然而,它们调控脂肪代谢的确切功能机制尚未完全阐明。在此,我们鉴定出了特异性获得包含4-氨基丁酸转氨酶(ABAT)的3'非翻译区(UTR)的miR-15a,并验证了其表达调控及其在脂肪生成机制中的作用。我们使用双荧光素酶报告基因检测以及在延边黄牛前体脂肪细胞中转染介导的miR-15a过表达和抑制,来研究miR-15a在脂肪生成中的作用。结果表明,miR-15a直接靶向ABAT的3'UTR并下调其表达。此外,在蛋白质和mRNA水平上,使用miRNA模拟物过表达miR-15a可抑制甘油三酯积累,并下调脂肪生成的过氧化物酶体增殖物激活受体γ和CCAAT增强子结合蛋白α,而抑制miR-15a则产生相反的效果。上述结果表明,miR-15a通过抑制ABAT的表达来调控延边黄牛前体脂肪细胞的分化。此外,我们的研究结果表明,miR-15a及其靶基因可能是研究牛肌肉内脂肪沉积和治疗人类肥胖症的新靶点。