晋南牛SREBP1基因调控前体脂肪细胞分化的研究  

Regulation of Preadipocyte Differentiation by SREBP 1 Gene in Jinnan Cattle

作　　者：张唯玉 程景[1] 许家宝 王静 陶薪燕 李博 张亚伟 张丹丹[1] 张宁[1] 郝振凯 周琛帛 张元庆 ZHANG Weiyu;CHENG Jing;XU Jiabao;WANG Jing;TAO Xinyan;LI Bo;ZHANG Yawei;ZHANG Dandan;ZHANG Ning;HAO Zhenkai;ZHOU Chenbo;ZHANG Yuanqing(College of Animal Science,Shanxi Agricultural University,Taigu 030801,China)

机构地区：[1]山西农业大学动物科学学院,太谷030801

出　　处：《畜牧兽医学报》2024年第11期5003-5017,共15页ACTA VETERINARIA ET ZOOTECHNICA SINICA

基　　金：山西省基础研究计划自然科学研究面上项目(20210302123424);肉牛边鸡地方特色畜禽种质资源创新及品种选育(202201140601026);山西省现代农业产业技术体系牛体系(2023CYJSTX13)。

摘　　要：旨在探究SREBP1基因对晋南牛前体脂肪细胞分化的影响,并通过RNA-Seq技术探究该基因在晋南牛前体脂肪细胞分化过程中的调控机制。本研究利用SREBP1基因的过表达载体和siRNA转染晋南牛前体脂肪细胞,油酸诱导分化后采用油红O染色观察脂滴累积情况,检测甘油三酯(triglyceride,TG)含量以探究SREBP1基因对其分化的影响。利用RT-qPCR和蛋白免疫印迹技术(Western blotting,WB)检测过表达SREBP1基因的前体脂肪细胞中相关基因mRNA和蛋白水平变化。RNA-Seq检测干扰SREBP1基因的脂肪细胞,筛选差异表达基因(differentially expressed genes,DEGs)进行KEGG通路富集分析,并对差异表达基因进行验证,以探究SREBP1基因参与调控晋南牛前体脂肪细胞分化相关的潜在靶基因。每个处理均设置3个重复。结果表明:1)过表达SREBP1基因能够促进前体脂肪细胞内脂滴的累积、极显著提高细胞内TG含量(P<0.01),干扰SREBP1基因则会抑制脂滴形成。2)过表达SREBP1基因后FABP4的mRNA表达量显著升高(P<0.05),FABP7、LPL的表达量极显著升高(P<0.01),WB结果显示FABP4蛋白表达水平明显升高;3)RNA-Seq共筛选到227个DEGs,其中包括67个上调基因和160个下调基因。京都基因和基因组百科全书(KEGG)富集分析结果表明差异基因富集到PPAR信号通路和不饱和脂肪酸生物合成等相关通路,干扰SREBP1基因后,差异表达基因FABP4、LPL和FABP7的mRNA表达量均极显著降低(P<0.01),FABP4的蛋白表达水平也明显降低。由此可知,SREBP1基因对于晋南牛前体脂肪细胞分化具有促进作用,过表达该基因可促进细胞内TG累积,并且极有可能是通过对与脂质分化相关的PPAR信号通路和不饱和脂肪酸生物合成通路的调控实现的。本研究结果为探究SREBP1基因在晋南牛前体脂肪细胞分化过程中的调控机制提供理论参考。This study aimed to investigate the effect of the SREBP 1 gene on the differentiation of Jinnan cattle precursor adipocytes and to explore the regulatory mechanism of this gene in the differentiation of Jinnan cattle precursor adipocytes by RNA-Seq technology.We transfected Jinnan cattle preadipocytes with an overexpression plasmid of SREBP 1 gene and siRNA,and after differentiation induced by oleic acid,oil red O staining was used to observe the accumulation of lipid droplets,and triglyceride(TG)content was tested to explore the influence of SREBP 1 gene on their differentiation.RT-qPCR and Western blotting(WB)were used to detect changes at mRNA and protein levels of related genes in preadipocytes overexpressing the SREBP 1 gene.RNA-Seq was used to detect adipocytes that interfered with the SREBP 1 gene,screened differentially expressed genes(DEGs),performed enrichment analysis of the KEGG pathway,and verified the differentially expressed genes.The potential target genes of the SREBP 1 gene involved in regulating the differentiation of precursor adipocytes of Jinnan cattle were explored.Three replicates were set up for each treatment.The results showed as follows:1)Compared with the control group,overexpression of SREBP 1 gene could promote the accumulation of lipid droplets in precursor adipocytes and significantly increase the intracellular triglyceride content(P<0.01),knockdown of SREBP 1 gene could inhibit lipid droplet formation.2)FABP 4 mRNA expression significantly increased after overexpression of the SREBP 1 gene(P<0.05),the expression of FABP 7 and LPL were significantly increased(P<0.01),WB results showed that FABP4 protein expression level increased obviously;3)A total of 227 DEGs were detected by RNA-Seq,including 67 up-regulated genes and 160 down-regulated genes.The Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment analysis showed that differentially expressed genes were enriched into PPAR signaling pathway and unsaturated fatty acid biosynthesis pathway,the mRNA expression of differentially

关 键 词：前体脂肪细胞 SREBP1 诱导分化 RNA-SEQ PPAR 

分 类 号：S823.2[农业科学—畜牧学]