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脂肪酸延长酶 7 通过 SP1 调控,参与牛乳腺上皮细胞的脂类积累。

Fatty acid elongase7 is regulated via SP1 and is involved in lipid accumulation in bovine mammary epithelial cells.

机构信息

Shaanxi Key Laboratory of Molecular Biology for Agriculture, College of Animal Science and Technology, Northwest A&F University, Yangling, Shaanxi, P.R. China.

College of Veterinary Medicine, Northwest A&F University, Yangling, Shaanxi, P.R. China.

出版信息

J Cell Physiol. 2018 Jun;233(6):4715-4725. doi: 10.1002/jcp.26255. Epub 2018 Jan 15.

Abstract

Fatty Acid Elongase 7 (ELOVL7) is the newly discovered protein on human that catalyzes the rate-limiting step towards the synthesis of very long-chain fatty acids and exhibits the highest activity toward C18: 3 (n-3) acyl-CoAs, which is the precursor of eicosapentaenoic acid (EPA, 20: 5n-3). However, in ruminants, an overall understanding of ELOVLs gene family and the transcriptional regulation of ELOVL7 remain unknown. The purpose of this study is to investigate the transcriptional regulation and the influence of bovine ELOVL7 in bovine mammary epithelial cells (bMECs). Quantitative real-time PCR analysis demonstrated that ELOVLs gene family had differential expression patterns in bMECs, and bovine ELOVL7 was expressed in a tissue-specific manner, which was high in kidney, followed by in abdominal fat and in bMECs. Promoter analysis of bovine ELOVL7, including bioinformatics analyzes, dual-luciferase reporter assays, protein pull-down assay, Western blot assay, over-expression and RNA interference assay, have independently and synthetically demonstrated that transcription factor Sp1 (SP1) specifically interacted with the GC-box at -143 to -128 base pair on ELOVL7 promoter. Furthermore, the exogenous α-linolenic acid (ALA, 18: 3n-3), strengthened the binding of SP1 to the ELOVL7 proximal promoter, resulting in the accumulation of lipid droplets in bMECs. In conclusion, these data suggest that the transcription of bovine ELOVL7 is affected by the binding of SP1 and the treatment of ALA, moreover, enlightening us the profound role of SP1 in modulating lipid synthesis of the mammary gland in cattle.

摘要

脂肪酸延长酶 7(ELOVL7)是在人体中发现的一种新蛋白,它催化长链脂肪酸合成的限速步骤,并对 C18:3(n-3)酰基辅酶 A 表现出最高的活性,这是二十碳五烯酸(EPA,20:5n-3)的前体。然而,在反刍动物中,对 ELOVLs 基因家族的整体理解和 ELOVL7 的转录调控仍不清楚。本研究旨在探讨转录调控和牛 ELOVL7 对牛乳腺上皮细胞(bMECs)的影响。定量实时 PCR 分析表明,ELOVLs 基因家族在 bMECs 中表现出不同的表达模式,牛 ELOVL7 呈组织特异性表达,在肾脏中表达最高,其次是腹部脂肪和 bMECs。牛 ELOVL7 的启动子分析,包括生物信息学分析、双荧光素酶报告基因检测、蛋白 pull-down 实验、Western blot 分析、过表达和 RNA 干扰实验,独立且综合地证明了转录因子 Sp1(SP1)特异性地与 ELOVL7 启动子上-143 到-128 碱基的 GC 盒相互作用。此外,外源性 α-亚麻酸(ALA,18:3n-3)增强了 SP1 与 ELOVL7 近端启动子的结合,导致 bMECs 中脂质滴的积累。总之,这些数据表明,牛 ELOVL7 的转录受 SP1 结合和 ALA 处理的影响,此外,还启发了我们 SP1 在调节牛乳腺脂质合成中的深刻作用。

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