Wuxi Fisheries College, Nanjing Agricultural University, Wuxi 214081, China; Key Laboratory of Freshwater Fisheries and Germplasm Resources Utilization, Ministry of Agriculture, Freshwater Fisheries Research Center, Chinese Academy of Fishery Sciences, Wuxi 214081, China.
Key Laboratory of Freshwater Fisheries and Germplasm Resources Utilization, Ministry of Agriculture, Freshwater Fisheries Research Center, Chinese Academy of Fishery Sciences, Wuxi 214081, China.
Gene. 2018 Jun 20;660:1-7. doi: 10.1016/j.gene.2018.03.064. Epub 2018 Mar 21.
MicroRNAs (miRNAs) are non-coding RNAs that function as post-transcriptional gene regulators and that play vital roles controlling lipid metabolism. miR-205 is an important miRNA related to adipogenesis and lipid metabolism. However, little is known about the potential role of miR-205-5p in genetically improved farmed tilapia (GIFT, Oreochromis niloticus). In this study, we used miRanda software to search for potential miR-205-5p target genes and found a lipid-metabolism-related gene called acetyl-CoA carboxylase β (ACACβ). Quantitative real-time polymerase chain reaction data indicated that there may be a negative regulation relationship between miR-205-5p and ACACβ gene expression under HFD rearing. Using luciferase reporter assays, we verified the binding site of miR-205-5p in the 3'-untranslated region of the ACACβ mRNA. Furthermore, an in vivo functional analysis of miR-205-5p was performed by injecting GIFT juveniles with a miR-205-5p antagomir. Reduced levels of miR-205-5p in GIFT liver increased ACACβ mRNA expression 12 h post-injection. miR-205-5p suppression also increased fatty acid synthase and peroxisome proliferator-activated receptor-α mRNA levels 48 h and 120 h post-injection, respectively. Taken together, our results indicate that miR-205-5p negative regulates hepatic ACACβ mRNA expression, and may serve as an important regulator in controlling hepatic lipid metabolism in GIFT.
微小 RNA（miRNA）是一类非编码 RNA，作为转录后基因调控因子发挥作用，在控制脂质代谢方面发挥着重要作用。miR-205 是一种与脂肪生成和脂质代谢相关的重要 miRNA。然而，关于 miR-205-5p 在遗传改良养殖罗非鱼（GIFT，Oreochromis niloticus）中的潜在作用知之甚少。在这项研究中，我们使用 miRanda 软件搜索潜在的 miR-205-5p 靶基因，发现了一个与脂质代谢相关的基因，称为乙酰辅酶 A 羧化酶 β（ACACβ）。定量实时聚合酶链反应数据表明，在高脂肪饮食饲养下，miR-205-5p 和 ACACβ 基因表达之间可能存在负调控关系。通过荧光素酶报告基因检测，我们验证了 miR-205-5p 在 ACACβ mRNA 3'-非翻译区的结合位点。此外，通过向 GIFT 幼鱼注射 miR-205-5p 拮抗剂，对 miR-205-5p 进行了体内功能分析。GIFT 肝脏中 miR-205-5p 水平降低可使 ACACβ mRNA 表达在注射后 12 小时增加。miR-205-5p 抑制还分别使脂肪酸合酶和过氧化物酶体增殖物激活受体-α mRNA 水平在注射后 48 小时和 120 小时增加。综上所述，我们的结果表明，miR-205-5p 负调控肝 ACACβ mRNA 表达，可能作为控制 GIFT 肝脂质代谢的重要调节因子。
