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circ01592 通过吸附 miR-218 调控奶牛乳腺上皮细胞不饱和脂肪酸代谢。

circ01592 regulates unsaturated fatty acid metabolism through adsorbing miR-218 in bovine mammary epithelial cells.

机构信息

College of Animal Science and Technology, Yangzhou University, Yangzhou 225009, P. R. China.

Joint International Research Laboratory of Agriculture & Agri-Product Safety, Ministry of Education, Yangzhou University, Yangzhou, China.

出版信息

Food Funct. 2021 Nov 29;12(23):12047-12058. doi: 10.1039/d1fo02797b.

Abstract

The composition of fatty acids plays a key role in regulating milk flavor and quality. Therefore, to improve the quality of milk, it is particularly important to study the regulatory mechanism of fatty acid metabolism in dairy cows. In this study, the expression profiles at non-lactation, peak-lactation, mid-lactation and late-lactation were constructed by high-throughput sequencing. Considering non-lactation as the control group and the other points as the experimental groups, the differentially expressed genes were screened. ELOVL5 was significantly upregulated and was selected for subsequent analyses. Bioinformatics prediction, a dual-luciferase assay, qPCR analysis and western blot analysis were used for verification. The results showed that ELOVL5 was a downstream target gene of miR-218 that regulated milk fat metabolism. A dual-luciferase assay and expression level analysis showed that circ01592 can directly bind to miR-218 and that overexpression of circ01592 (pcDNA-circ01592) significantly reduced the expression of miR-218 and enhanced the expression of ELOVL5, the target gene of miR-218 in BMECs. A functional study of BMECs showed that circ01592 promoted the synthesis of TAG and increased the content of UFA. The function of miR-218 was opposite to that of circ01592. Overall, the data showed that circ01592 promoted TAG synthesis and fatty acid composition by binding miR-218, alleviating the inhibitory effect of miR-218 on ELOVL5 expression. These mechanisms provide a new research approach and theoretical basis for improving milk quality.

摘要

脂肪酸的组成在调节乳味和品质方面起着关键作用。因此,为了提高牛奶的质量,研究奶牛脂肪酸代谢的调节机制尤为重要。本研究通过高通量测序构建了非泌乳期、泌乳高峰期、泌乳中期和泌乳后期的表达谱,以非泌乳期为对照组,其他点为实验组,筛选差异表达基因。ELOVL5 显著上调,被选为后续分析。采用生物信息学预测、双荧光素酶报告基因检测、qPCR 分析和 Western blot 分析进行验证。结果表明,ELOVL5 是 miR-218 调节乳脂代谢的下游靶基因。双荧光素酶报告基因检测和表达水平分析表明,circ01592 可直接与 miR-218 结合,circ01592 的过表达(pcDNA-circ01592)显著降低了 miR-218 的表达,增强了 miR-218 的靶基因 ELOVL5 在 BMECs 中的表达。BMECs 的功能研究表明,circ01592 促进 TAG 的合成,增加 UFA 的含量。miR-218 的功能与 circ01592 相反。综上所述,数据表明 circ01592 通过与 miR-218 结合促进 TAG 合成和脂肪酸组成,缓解 miR-218 对 ELOVL5 表达的抑制作用。这些机制为改善牛奶质量提供了新的研究方法和理论依据。

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