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

作　　者：胡濒月 胡杨 成文敏 赵素梅[2] 赵红业[1,3,4] 魏红江 HU Bin-yue;HU Yang;CHENG Wen-min;ZHAO Su-mei;ZHAO Hong-Ye;WEI Hong-Jiang(Key Laboratory of Animal Gene Editing and Cell Cloning in Yunnan Province,Kunming 650201;College of Animal Science and Technology,Yunnan Agricultural University,Kunming 650201;College of Plant Protection,Yunnan Agricultural University,Kunming 650201;State Key Laboratory for Conservation and Utilization of Bio-Resources in Yunnan,Kunming 650201)

机构地区：[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

出　　处：《生物技术通报》2020年第8期111-119,共9页Biotechnology Bulletin

基　　金：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)。

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。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 PPARγ,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.

关 键 词：LEPTIN 猪前脂肪细胞 脂滴 脂质合成代谢相关基因 

分 类 号：S828[农业科学—畜牧学]