过表达UCP3基因对萨湖羊前体脂肪细胞分化的影响  

Effect of Overexpression of UCP3 Gene on the Differentiation of Sahu Sheep Preadipocytes

作　　者：刘林丽[2] 彭雪兰 李博 程乐凡 次仁拉姆 张恩平[1] LIU Linli;PENG Xuelan;LI Bo;CHENG Lefan;CIREN Lamu;ZHANG Enping(College of Animal Science and Technology,Northwest A&F University,Yangling 712100,China;College of Life Science,Northwest A&F University,Yangling 712100,China)

机构地区：[1]西北农林科技大学动物科技学院,杨凌712100 [2]西北农林科技大学生命科学学院,杨凌712100

出　　处：《畜牧兽医学报》2023年第8期3275-3285,共11页ACTA VETERINARIA ET ZOOTECHNICA SINICA

基　　金：陕西省肉羊产业技术体系建设项目(NYKJ-2020-YL-14,NYKJ-2021-YL(XN)43)。

摘　　要：旨在阐明萨湖羊解偶联蛋白3(UCP3)基因的组织、细胞表达模式,阐明过表达UCP3对揭示对萨湖羊前体脂肪细胞分化的影响。本研究通过实时荧光定量PCR(real-time quantitative PCR,qPCR)技术检测UCP3基因在3只萨湖羊心脏、肝脏、脾脏等组织中的表达分布及在成脂诱导分化不同时期的表达水平;过表达UCP3后,采用qPCR验证过表达效率;利用油红O染色、脂质提取分析及脂肪细胞分化标志基因的检测,分别从细胞形态学和生物学角度明确过表达UCP3对萨湖羊脂肪细胞脂质生成的影响。结果表明:UCP3基因在萨湖羊各组织中广泛表达,在心脏和脾脏中相对表达量较高;时序表达谱显示,在萨湖羊前体脂肪细胞分化过程中UCP3基因表达量呈上升的趋势(P<0.05);过表达UCP3后,萨湖羊前体脂肪细胞的脂滴明显增多,极显著增加了脂质含量(P<0.01);脂肪细胞分化标志基因PPARγ、C/EBPα、LEP mRNA水平极显著上升(P<0.01),FAS、UCP1 mRNA水平显著上升(P<0.05)。本研究揭示了萨湖羊UCP3的表达模式,验证了过表达UCP3能够促进萨湖羊前体脂肪细胞的成脂分化,推测UCP3是萨湖羊前体脂肪细胞的正调控因子,可能是通过调控PPARγ、C/EBPα和FAS等分化关键基因的表达来实现的。The purpose of this study was to clarify the tissue and cell expression pattern of uncoupling protein 3(UCP3)gene in Sahu sheep,and to clarify the effect of overexpression of UCP3 on revealing the differentiation of preadipocytes in Sahu sheep.This study used real-time quantitative PCR(qPCR)technology to detect the expression distribution of UCP3 gene in the heart,liver,spleen and other tissues of 3 Sahu sheep,as well as its expression levels at different stages of adipogenic differentiation;After overexpression of UCP3,qPCR was used to verify the overexpression efficiency,using Oil Red O staining,lipid extraction analysis,and adipocyte differentiation marker genes testing,clarify the effects of overexpression of UCP3 on lipid production in Sahu sheep preadipocytes from both cellular morphology and biological perspectives.The results showed that the UCP3 gene was widely expressed in various tissues of Sahu sheep,with relatively higher expression levels in the heart and spleen;The temporal expression profile showed that the expression level of UCP3 gene showed an upward trend during the differentiation of preadipocytes in Sahu sheep(P<0.05);After overexpression of UCP3,the number of lipid droplets in the preadipocytes of Sahu sheep increased significantly(P<0.01);In addition,the mRNA levels of PPARγ,C/EBPαand LEP were extremely significantly up-regulated(P<0.01),and FAS and UCP 1 mRNA levels were significantly up-regulated(P<0.05).This study revealed the expression pattern of UCP3 in Sahu sheep and verified that overexpression of UCP3 can promote adipogenic differentiation of Sahu sheep preadipocytes.It is speculated that UCP3 is a positive regulatory factor of Sahu sheep preadipocytes,it possibly regulates the differentiation of preadipocytes through regulating the expression of PPARγ,C/EBPαand FAS genes.

关 键 词：萨湖羊 前体脂肪细胞 pcDNA3.1-flag UCP3 无缝克隆 

分 类 号：S826.2[农业科学—畜牧学]