College of Animal Science and Veterinary Medicine, Henan Agricultural University, Zheng Zhou 450002, Henan, China.
Int J Mol Sci. 2019 Aug 20;20(16):4063. doi: 10.3390/ijms20164063.
Our previous studies showed that microRNA-15a (miR-15a) was closely related to intramuscular fat (IMF) deposition in chickens; however, its regulatory mechanism remains unclear. Here, we evaluated the expression characteristics of miR-15a and its relationship with the expression of acetyl-CoA acyltransferase 1 (), acyl-CoA oxidase 1 () and sterol carrier protein 2 () by qPCR analysis in Gushi chicken breast muscle at 6, 14, 22, and 30 weeks old, where we performed transfection tests of miR-15a mimics in intramuscular preadipocytes and verified the target gene of miR-15a in chicken fibroblasts (DF1). The miR-15a expression level at 30 weeks increased 13.5, 4.5, and 2.7-fold compared with the expression levels at 6, 14, and 22 weeks, respectively. After 6 days of induction, miR-15a over-expression significantly promoted intramuscular adipogenic differentiation and increased cholesterol and triglyceride accumulation in adipocytes. Meanwhile, 48 h after transfection with miR-15a mimics, the expression levels of , and genes decreased by 56.52%, 31.18% and 37.14% at the mRNA level in intramuscular preadipocytes. In addition, the co-transfection of miR-15a mimics and , and 3'UTR (untranslated region) dual-luciferase vector significantly inhibited dual-luciferase activity in DF1 cells. Taken together, our data demonstrate that miR-15a can reduce fatty acid oxidation by targeting , , and , which subsequently indirectly promotes the differentiation of chicken intramuscular preadipocytes.
我们之前的研究表明,microRNA-15a(miR-15a)与鸡的肌内脂肪(IMF)沉积密切相关;然而,其调控机制尚不清楚。在这里,我们通过 qPCR 分析评估了 miR-15a 在固始鸡胸肌中的表达特征及其与乙酰辅酶 A 酰基转移酶 1()、酰基辅酶 A 氧化酶 1()和固醇载体蛋白 2()表达的关系,在 6、14、22 和 30 周龄时,我们在肌内前体脂肪细胞中转染 miR-15a 模拟物,并在鸡成纤维细胞(DF1)中验证了 miR-15a 的靶基因。与 6、14 和 22 周龄时的表达水平相比,30 周龄时 miR-15a 的表达水平分别增加了 13.5、4.5 和 2.7 倍。诱导 6 天后,miR-15a 过表达显著促进肌内脂肪生成分化,并增加脂肪细胞中胆固醇和甘油三酯的积累。同时,在转染 miR-15a 模拟物 48 小时后,肌内前体脂肪细胞中、和基因的表达水平分别下降了 56.52%、31.18%和 37.14%。此外,miR-15a 模拟物与、和 3'UTR(非翻译区)双荧光素酶载体的共转染显著抑制了 DF1 细胞中的双荧光素酶活性。综上所述,我们的数据表明,miR-15a 可以通过靶向、和,间接减少脂肪酸氧化,从而促进鸡肌内前体脂肪细胞的分化。