摘要
为了研究固醇调节元件结合蛋白(sterol regulatory element binding factor 1,SREBP1)基因的表达与细胞脂肪形成的关系,本研究构建了pEGFP-C1-SREBP1重组质粒,利用脂质体介导法将重组质粒pEGFP-C1-SREBP1及pEGFP-C1空质粒转染至牛胎儿骨骼肌成纤维细胞培养48h,实时荧光定量PCR和Western blotting分析目标基因表达水平的变化,并采用油红O染色检测细胞内脂滴。结果发现,pEGFP-C1-SREBP1重组质粒转染细胞内SREBP1mRNA的表达水平显著高于空白对照组和阴性对照组(P<0.05);SREBP1蛋白也出现明显上调;空白对照组和pEGFP-C1阴性对照组细胞内均未见有红染细胞,而pEGFP-C1-SREBP1重组质粒转染组中出现了清晰的红染细胞。说明SREBP1的表达可促进牛胎儿骨骼肌成纤维细胞脂肪合成,证实牛胎儿成纤维细胞中SREBP1表达和脂滴形成之间存在直接关系。
The objective of this study was to investigate the relationship between sterol regulatory element binding factor 1（SREBP1） gene expression and cell fat formation,recombinant plasmid pEGFP-C1-SREBP1 was constructed and transferred into bovine fetal fibroblasts with Lipofectamine 2000,and then the cells were cultured for 48 h.Furthermore,the expressions of SREBP1 mRNA and protein were detected by Real-time PCR and Western blotting.Oil red O staining was used to explore cellular lipid droplet formation.The results showed that an up-regulation of SREBP1 mRNA in recombinant plasmid transfection group compared with normal control group and pEGFP-C1 transfected group,SREBP1 protein also appeared obvious rise.Oil red O found no lipid droplet in normal control group and pEGFP-C1 transfected group,but marked lipid droplets were detected in pEGFP-C1-SREBP1 transfected group.In conclusion,the up-regulation of SREBP1 directly resulteds in cellular lipid droplet formation in bovine fetal fibroblasts.
出处
《中国畜牧兽医》
CAS
北大核心
2013年第5期133-139,共7页
China Animal Husbandry & Veterinary Medicine
基金
“十二五”国家科技支撑计划“优质肉牛新品种(系)选育与关键技术研究及示范(2011BAD28B04)”
所基本科研业务费专项资金项目“基于高密度SNP建立中国肉牛群体全基因组选择优化方案(2010jc-2)”
农业部专项“现代农业(肉牛)产业技术体系岗位科学家(CARS-38)”
关键词
固醇调节元件结合蛋白
过表达
牛胎儿成纤维细胞
脂滴形成
质粒转染
sterol regulatory element binding factor 1
overexpression
bovine fetal fibroblasts
lipid droplet formation
plasmid transfection
