MicroRNAs (miRNAs) play an important role in regulating gene expression, and myostatin (MSTN) has been widely recognized as a key gene for muscle growth and development. Through high-throughput sequencing to study the effects of starvation on miRNA transcriptomes in Larimichthys crocea muscle tissue, we found that the expression of miR-2014, miR-1231 and miR-1470 were significantly different between fasting and normal feeding Larimichthys crocea. Bioinformatics analysis predicted that miR-2014, miR-1231 and miR-1470 target MSTN mRNA 3’UTR. To verify the accuracy of predictions, we constructed double luciferase plasmids containing MSTN 3’UTR and confirmed that miR-2014-5p and miR-1231-5p can inhibit MSTN expression by targeting MSTN 3’UTR using double luciferase experiments, while miR-1470 is not involved in regulation. Subsequent site-directed mutation experiments reflected the specificity of the target sequence. In addition, quantitative PCR experiments revealed that miR-2014-5p and miR-1231-5p may participate in the regulation of MSTN expression in fasting and refeeding period, respectively. These results implied that miRNA may take part in muscle growth regulation during starvation. It provides some insights into the molecular regulation mechanism of MSTN in response to starvation stress in fish.

微小RNA（miRNA）在调节基因表达中起重要作用，而肌肉生长抑制素（MSTN）已被广泛认为是肌肉生长发育的关键基因。通过高通量测序研究饥饿对Larimichthys crocea肌肉组织中miRNA转录组的影响，我们发现禁食和正常喂养的Larimichthys crocea之间的miR-2014，miR-1231和miR-1470的表达存在显着差异。生物信息学分析预测，miR-2014，miR-1231和miR-1470靶向MSTN mRNA 3'UTR。为了验证预测的准确性，我们构建了包含MSTN 3'UTR的双重荧光素酶质粒，并确认miR-2014-5p和miR-1231-5p可以抑制MSTN表达通过靶向MSTN使用双荧光素酶实验3'UTR，而的miR-1470未在参与调节。随后的定点突变实验反映了靶序列的特异性。此外，定量PCR实验表明，miR-2014-5p和miR-1231-5p可能分别在禁食期和补料期参与MSTN表达的调控。这些结果表明，miRNA可能在饥饿期间参与肌肉生长调节。它提供了对MSTN响应鱼类饥饿压力的分子调控机制的一些见解。