College of Animal Science and Technology, Henan Agricultural University, Zhengzhou, Henan Province, 450002, P. R. China.
Mol Biol Rep. 2021 Jan;48(1):491-502. doi: 10.1007/s11033-020-06080-4. Epub 2021 Jan 4.
Our previous studies have shown that miR-125b-5p was highly expressed and significantly upregulated during abdominal fat deposition in chickens. However, the role of miR-125b in the regulation of adipogenesis is not clear in chickens. Therefore, we evaluated the effects of miR-125b-5p on preadipocyte proliferation and differentiation and the interaction between miR-125b-5p and the acyl-CoA synthetase bubblegum family member 2 (ACSBG2) gene in adipogenesis in chicken abdominal adipose tissue. Here, transfection tests of miR-125b-5p mimic/inhibitor were performed in preadipocytes, and the effects of miR-125b-5p on preadipocytes proliferation and differentiation were analyzed. The target site of miR-125b-5p in the 3'UTR (untranslated region) of ACSBG2 were verified by a luciferase reporter assay. Our results showed that miR-125b-5p overexpression inhibited proliferation and reduced the number of cells in S phase and G2/M phase in preadipocytes; conversely, miR-125b-5p inhibition promoted the proliferation and increased the number of cells in S phase and G2/M phase. In adipocytes after induction, miR-125b-5p overexpression led to a notable increase in the accumulation of lipid droplets as well as in the concentration of triglycerides, while miR-125b-5p inhibition had the opposite effect. Furthermore, miR-125b-5p could directly bind to the 3'UTR of ACSBG2, and its overexpression could significantly repress the mRNA and protein expression of ACSBG2. These results indicate that miR-125b-5p can inhibit preadipocyte proliferation and can promote preadipocyte differentiation to affect adipogenesis in chicken abdominal adipose tissues, at least partially by downregulating ACSBG2.
我们之前的研究表明,miR-125b-5p 在鸡腹部脂肪沉积过程中高度表达且显著上调。然而,miR-125b 在鸡脂肪生成中的调节作用尚不清楚。因此,我们评估了 miR-125b-5p 对鸡腹部脂肪组织前体脂肪细胞增殖和分化的影响,以及 miR-125b-5p 与酰基辅酶 A 合成酶泡泡糖家族成员 2(ACSBG2)基因在脂肪生成中的相互作用。在这里,我们在前体脂肪细胞中进行了 miR-125b-5p 模拟物/抑制剂的转染试验,并分析了 miR-125b-5p 对前体脂肪细胞增殖和分化的影响。通过荧光素酶报告基因试验验证了 miR-125b-5p 在 ACSBG2 的 3'UTR(非翻译区)中的靶位点。结果表明,miR-125b-5p 过表达抑制增殖并减少前体脂肪细胞 S 期和 G2/M 期的细胞数量;相反,miR-125b-5p 抑制促进增殖并增加 S 期和 G2/M 期的细胞数量。在诱导后的脂肪细胞中,miR-125b-5p 过表达导致脂滴积累显著增加,甘油三酯浓度也升高,而 miR-125b-5p 抑制则产生相反的效果。此外,miR-125b-5p 可以直接结合 ACSBG2 的 3'UTR,其过表达可以显著抑制 ACSBG2 的 mRNA 和蛋白表达。这些结果表明,miR-125b-5p 可以抑制前体脂肪细胞增殖,促进前体脂肪细胞分化,从而影响鸡腹部脂肪组织的脂肪生成,至少部分是通过下调 ACSBG2 实现的。