Leptin过表达对猪前脂肪细胞脂滴形成的研究

doi:10.13560/j.cnki.biotech.bull.1985.2020-0072

生物技术通报 ›› 2020, Vol. 36 ›› Issue (8): 111-119.doi: 10.13560/j.cnki.biotech.bull.1985.2020-0072

Leptin过表达对猪前脂肪细胞脂滴形成的研究

胡濒月^1,2,4, 胡杨^1,3, 成文敏^1,2,4, 赵素梅², 赵红业^1,3,4, 魏红江^1,4

1.云南省动物基因编辑与体细胞克隆技术重点实验室,昆明 650201;
2.云南农业大学动物科学技术学院,昆明 650201;
3.云南农业大学植物保护学院,昆明 650201;
4.云南生物资源保护与利用国家重点实验室,昆明 650201

收稿日期:2020-01-19 出版日期:2020-08-26 发布日期:2020-08-27
作者简介:胡濒月,女,硕士,研究方向：动物分子营养与代谢调控;E-mail：binyue_hu@163.com;胡杨同为本文第一作者
基金资助:
云南省生物医药重大研发平台构建（2018ZF018）,云南省应用基础研究计划重点项目（2016FA010）,云南省院士（专家）工作站（2018IC064）

Lipid Droplet Formation in the Pre-adipocytes of Leptin-overexpressed Pig

HU Bin-yue^1,2,4, HU Yang^1,3, CHENG Wen-min^1,2,4, ZHAO Su-mei², ZHAO Hong-Ye^1,3,4, WEI Hong-Jiang^1,4

1. Key Laboratory of Animal Gene Editing and Cell Cloning in Yunnan Province,Kunming 650201;
2. College of Animal Science and Technology,Yunnan Agricultural University,Kunming 650201;
3. College of Plant Protection,Yunnan Agricultural University,Kunming 650201;
4. State Key Laboratory for Conservation and Utilization of Bio-Resources in Yunnan,Kunming 650201

Received:2020-01-19 Published:2020-08-26 Online:2020-08-27

摘要/Abstract

摘要： 研究Leptin过表达对猪前脂肪细胞脂滴形成的影响,旨为进一步研究Leptin与脂质代谢相关的分子机制奠定理论基础。选取Leptin过表达与野生型猪皮下脂肪组织,在无菌条件下分离前脂肪细胞进行传代培养并诱导分化形成脂滴,通过油红O和Bodipy染色后观察脂滴面积并分析脂质的含量,利用Q-PCR检测脂滴形成相关基因mRNA的表达水平。诱导4 d后可分化形成脂滴,油红O和Bodipy染色的结果显示,Leptin过表达猪前脂肪细胞脂滴数量和甘油三酯含量显著低于野生型（P<0.05）;且脂质合成相关基因PPARγ、SCAP、SREPB1和PLIN2的表达水平显著低于野生型（P<0.01）。结果表明,过表达Leptin可促使猪前脂肪细胞中PPARγ、SREPB1、SCAP、PLIN2基因的表达下调,进而抑制脂滴形成。

关键词: leptin, 猪前脂肪细胞, 脂滴, 脂质合成代谢相关基因

Abstract: This work aims to investigate the effect of endogenous leptin overexpression on lipid droplet formation in pig adipocytes and to provide a theoretical basis for studying the molecular mechanism of Leptin regulating lipid metabolism. The subcutaneous adipocytes of leptin-overexpressed and wild-type pig were isolated and cultured under aseptic condition. Then these cells were induced to be differentiated to produce lipid droplets（LDs）. The area of a lipid droplet and the content of the lipid were observed by microscope after Oil Red O and Bodipy staining. Then the expression levels of lipid droplet formation-related genes were determined by Q-PCR. The cells differentiated to form LDs after 4 d of induction. The results from the Oil Red O and Bodipy staining showed that the number of lipid droplets and lipid contents in the leptin-overexpressed porcine adipocytes were significantly lower than those in the wild type（P<0.05）. Furthermore,the expression levels of lipogenesis-related genes PPAR γ,SCAP,SREPB1 and PLIN2 in the leptin-overexpressed porcine adipocytes were significantly lower compared to the wild type（P<0.01）. Leptin overexpression inhibited the formation of LDs by down-regulating PPARγ,SREPB1,SCAP and PLIN2 genes expression in porcine pre-adipocytes.

Key words: leptin, porcine adipocytes, lipid droplets, lipogenesis and metabolism-related genes

胡濒月, 胡杨, 成文敏, 赵素梅, 赵红业, 魏红江. Leptin过表达对猪前脂肪细胞脂滴形成的研究[J]. 生物技术通报, 2020, 36(8): 111-119.

HU Bin-yue, HU Yang, CHENG Wen-min, ZHAO Su-mei, ZHAO Hong-Ye, WEI Hong-Jiang. Lipid Droplet Formation in the Pre-adipocytes of Leptin-overexpressed Pig[J]. Biotechnology Bulletin, 2020, 36(8): 111-119.

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Leptin过表达对猪前脂肪细胞脂滴形成的研究

Lipid Droplet Formation in the Pre-adipocytes of Leptin-overexpressed Pig

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