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研究和的表达分析及其转录因子对牛脂肪细胞分化的协调调控。

Study of expression analysis of and the coordinate regulation of bovine adipocyte differentiation by and its transcription factors.

机构信息

College of Animal Science and Technology, Northwest A&F University, Yangling 712100, Shaanxi, China.

National Beef Cattle Improvement Center, Northwest A&F University, Yangling 712100, Shaanxi, China.

出版信息

Biosci Rep. 2018 Dec 14;38(6). doi: 10.1042/BSR20181705. Print 2018 Dec 21.

DOI:10.1042/BSR20181705
PMID:30442871
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC6294651/
Abstract

Sirtuins, NAD-dependent deacylases and ADP-ribosyltransferases, are critical regulators of metabolism involved in many biological processes, and are involved in mediating adaptive responses to the cellular environment. SIRT4 is a mitochondrial sirtuin and has been shown to play a critical role in maintaining insulin secretion and glucose homeostasis. As a regulator of lipid homeostasis, SIRT4 can repress fatty acid oxidation and promote lipid anabolism in nutrient-replete conditions. Using real-time quantitative PCR (qPCR) to explore the molecular mechanisms of transcriptional regulation of bovine during adipocyte differentiation, we found that bovine is expressed at high levels in bovine subcutaneous adipose tissue. knockdown led to decreased expression of adipogenic differentiation marker genes during adipocyte differentiation. The core promoter of bovine was identified in the -402/-60 bp region of the cloned 2-kb fragment containing the 5'-regulatory region. Binding sites were identified in this region for E2F transcription factor-1 (E2F1), CCAAT/enhancer-binding protein β (CEBPβ), homeobox A5 (HOXA5), interferon regulatory factor 4 (IRF4), paired box 4 (PAX4), and cAMP responsive element-binding protein 1 (CREB1) by using Electrophoretic mobility shift assay (EMSA) and luciferase reporter gene assay. We also found that E2F1, CEBPβ, and HOXA5 transcriptionally activate expression, whereas, IRF4, PAX4, and CREB1 transcriptionally repress expression. We further verified that knockdown could affect the ability of these transcription factors (TFs) to regulate the differentiation of bovine adipocytes. In conclusion, our results shed light on the mechanisms underlying the transcriptional regulation of expression in bovine adipocytes.

摘要

去乙酰化酶和 ADP-核糖基转移酶依赖 NAD 的 Sirtuins 是参与许多生物过程的代谢关键调节因子,并且参与介导细胞环境的适应性反应。SIRT4 是一种线粒体 Sirtuin,已被证明在维持胰岛素分泌和葡萄糖稳态中起关键作用。作为脂质稳态的调节剂,SIRT4 可以抑制脂肪酸氧化并促进营养丰富条件下的脂质合成。使用实时定量 PCR(qPCR)探索牛在脂肪细胞分化过程中转录调控的分子机制,我们发现牛在牛皮下脂肪组织中表达水平较高。在脂肪细胞分化过程中,沉默会导致脂肪生成分化标记基因的表达减少。克隆的 2-kb 片段包含 5'-调控区,在该克隆的-402/-60 bp 区域鉴定出牛的核心启动子。在该区域鉴定出 E2F 转录因子-1(E2F1)、CCAAT/增强子结合蛋白β(CEBPβ)、同源盒 A5(HOXA5)、干扰素调节因子 4(IRF4)、配对盒 4(PAX4)和 cAMP 反应元件结合蛋白 1(CREB1)的结合位点通过使用电泳迁移率变动分析(EMSA)和荧光素酶报告基因分析。我们还发现 E2F1、CEBPβ 和 HOXA5 转录激活表达,而 IRF4、PAX4 和 CREB1 转录抑制表达。我们进一步验证了沉默可以影响这些转录因子(TFs)调节牛脂肪细胞分化的能力。总之,我们的结果阐明了牛脂肪细胞中表达的转录调控机制。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cd2c/6294651/3238e18aafd1/bsr-38-bsr20181705-g6.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cd2c/6294651/cb85dda2ef08/bsr-38-bsr20181705-g1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cd2c/6294651/5dfdd7bb79f2/bsr-38-bsr20181705-g2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cd2c/6294651/879b0a4feb52/bsr-38-bsr20181705-g3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cd2c/6294651/8c0782955346/bsr-38-bsr20181705-g4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cd2c/6294651/021281bac174/bsr-38-bsr20181705-g5.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cd2c/6294651/3238e18aafd1/bsr-38-bsr20181705-g6.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cd2c/6294651/cb85dda2ef08/bsr-38-bsr20181705-g1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cd2c/6294651/5dfdd7bb79f2/bsr-38-bsr20181705-g2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cd2c/6294651/879b0a4feb52/bsr-38-bsr20181705-g3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cd2c/6294651/8c0782955346/bsr-38-bsr20181705-g4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cd2c/6294651/021281bac174/bsr-38-bsr20181705-g5.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cd2c/6294651/3238e18aafd1/bsr-38-bsr20181705-g6.jpg

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