miRNA-224 通过靶向 LPL 抑制牛前体脂肪细胞的成脂分化。

MicroRNA-224 impairs adipogenic differentiation of bovine preadipocytes by targeting LPL.

机构信息

Animal Husbandry and Veterinary Research Institute, Shanghai Academy of Agricultural Sciences, Shanghai, China.

College of Animal Science and Technology, National Beef Cattle Improvement Center, Northwest A&F University, Yangling, Shaanxi, China.

出版信息

Mol Cell Probes. 2019 Apr;44:29-36. doi: 10.1016/j.mcp.2019.01.005. Epub 2019 Jan 28.

DOI:10.1016/j.mcp.2019.01.005

PMID:30703449

Abstract

In this study, the potential functions of miR-224 in regulating adipogenic differentiation were explored in bovine preadipocytes. Comparative transcriptome analysis between castrated male cattle with increased intramuscular fat (IMF) and intact male cattle revealed that miR-224 and LPL were abnormally expressed, correlating negatively, and LPL was a predicted target of miR-224. A dual luciferase reporter assay confirmed the negative targeting regulatory relationship between miR-224 and LPL. When miR-224 was either overexpressed or silenced, qRT-PCR showed a negative regulatory effect on LPL. mRNA expression levels of the fat-formation-related biomarkers C/EBPα, C/EBPβ, PPARγ, FASN and PLIN1 decreased when miR-224 was overexpressed, while the opposite effect occurred and adipogenic differentiation followed when miR-224 was inhibited. Oil Red O staining. Triglyceride (TG) levels and immunostaining revealed that the accumulation of lipid droplets decreased or increased accordingly. Taken together, our data demonstrated that miR-224 regulated the adipogenic differentiation of bovine preadipocytes by targeting LPL. This provides insight into the molecular basis of IMF deposition in beef cattle.

摘要

在这项研究中，我们探讨了 miR-224 在调控牛前体脂肪细胞成脂分化中的潜在功能。对肌肉内脂肪（IMF）增加的去势公牛肉牛与完整公牛肉牛进行比较转录组分析，结果显示 miR-224 和 LPL 异常表达，呈负相关，LPL 是 miR-224 的预测靶标。双荧光素酶报告基因实验证实了 miR-224 与 LPL 之间的负向靶向调控关系。当 miR-224 过表达或沉默时，qRT-PCR 显示其对 LPL 具有负调控作用。当 miR-224 过表达时，脂肪形成相关生物标志物 C/EBPα、C/EBPβ、PPARγ、FASN 和 PLIN1 的 mRNA 表达水平降低，而当 miR-224 被抑制时，则出现相反的效果，脂肪细胞分化随之发生。油红 O 染色、三酰甘油（TG）水平和免疫染色显示，脂滴的积累相应减少或增加。综上所述，我们的数据表明，miR-224 通过靶向 LPL 调控牛前体脂肪细胞的成脂分化。这为了解肉牛 IMF 沉积的分子基础提供了线索。