NR1H3基因调控猪前体脂肪细胞分化的研究  

Study on NR1H3 Gene Regulating Differentiation of Porcine Preadipocyte

作　　者：史明月 张雪莲[1] 杨晓奋 牛瑾 邢建东 路畅 高鹏飞[1] 郭晓红[1] 李步高[1] 曹果清[1] SHI Mingyue;ZHANG Xuelian;YANG Xiaofen;NIU Jin;XING Jiandong;LU Chang;GAO Pengfei;GUO Xiaohong;LI Bugao;CAO Guoqing(College of Animal Science, Shanxi Agricultural University, Taigu 030801, China;The Center for Modern Agricultural Development, Bureau of Agriculture and Rural Affairs of Jincheng City, Jincheng 048000, China)

机构地区：[1]山西农业大学动物科学学院,太谷030801 [2]晋城市农业农村局现代农业发展中心,晋城048000

出　　处：《畜牧兽医学报》2022年第7期2094-2103,共10页ACTA VETERINARIA ET ZOOTECHNICA SINICA

基　　金：国家自然科学基金(31872336);三晋学者支持计划专项经费资助(2016,2017)。

摘　　要：旨在探讨NR1H3基因在猪脂肪组织中的发育性表达规律及对猪前体脂肪细胞成脂分化的影响,以确定其在脂肪沉积过程中的主要功能。本研究采用qRT-PCR方法检测30、90、240日龄马身猪皮下脂肪组织中NR1H3的发育性表达规律;采集5日龄杜长大仔猪背部脂肪组织,分离猪前体脂肪细胞;通过细胞免疫荧光技术检测细胞中Adiponectin含量以鉴定细胞的纯度;构建猪NR1H3基因的过表达载体,设计NR1H3 siRNA序列,分别转染分离得到的猪前体脂肪细胞,采用qRT-PCR、Western blot和油红O染色等方法检测过表达和干扰效率及它们对成脂分化关键基因表达的影响。结果表明,马身猪皮下脂肪组织中NR1H3的表达量随日龄增加呈上升趋势,30日龄时表达量最低,240日龄时表达量最高,差异极显著(P<0.01)。与对照组相比,猪前体脂肪细胞中过表达NR1H3,脂肪细胞的脂滴数明显增多,下游靶标SREBP-1c和ChREBP的表达量极显著提高(P<0.01),且成脂关键基因FAS、C/EBPβ、PPARγ和FABP4的mRNA表达量极显著提高(P<0.01),促进成脂过程;相反,干扰NR1H3基因,脂滴数明显减少,下游靶标及成脂关键基因的mRNA表达量极显著下调(P<0.01),抑制成脂过程。本研究表明,NR1H3基因是猪前体脂肪细胞成脂分化的正调节剂,通过影响其下游靶标SREBP-1c和ChREBP的表达而影响成脂分化,研究结果对阐明猪脂肪沉积的分子机理、改善肉质品质有重要意义。The purpose of this study was to investigate the developmental expression pattern of NR1H3 gene in subcutaneous adipose tissues of pigs at different ages and the effects of NR1H3 gene on adipogenic differentiation of porcine preadipocytes,and determine its primary function in fat deposition.qRT-PCR was used to detect the expression of NR1H3 in subcutaneous adipose tissues of Mashen pigs aged 30,90 and 240 days.Porcine preadipocytes were isolated from dorsal subcutaneous adipose tissues of 5-day-old DLY(Duroc×(Landrace×Yorksire))piglets.Cellular immunofluorescence technique was used to detect the Adiponectin protein content in the cells to identify the purity of preadipocytes.The over-expression vector of porcine NR1H3 gene was constructed and the NR1H3 siRNA sequences were designed,and then both of them were transfected into porcine preadipocytes.The overexpression and interference efficiency and their effects on expression of key genes related to adipogenic differentiation were investigated by qRT-PCR,Western blot and Oil red O staining methods.The results showed that the expression of NR1H3 in subcutaneous adipose tissues increased as the age increased,the expression level was the highest at 240 days of age and the lowest at 30 days of age,with extremely significant difference between them(P<0.01).Compared with negative control(NC),over-expression(OE)of NR1H3 significantly increased the number of lipid droplets in adipocytes,the expression levels of downstream targets SREBP-1c and ChREBP extremely significantly increased(P<0.01),and the mRNA expression levels of key adipogenic genes FAS,C/EBPβ,PPARγand FABP4 extremely significantly increased(P<0.01).On the contrary,interfering with NR1H3 significantly reduced the number of lipid droplets.The mRNA expression of downstream target genes and key adipogenic genes extremely significantly reduced(P<0.01),which inhibited the adipogenesis process.The results showed that NR1H3 gene was a positive regulator of the adipogenic differen-tiation of porcine preadipocytes,a

关 键 词：猪 NR1H3 猪前体脂肪细胞 成脂分化 

分 类 号：S828.2[农业科学—畜牧学]