固醇调节元件结合蛋白基因的过表达对促进牛胎儿成纤维细胞脂肪形成的影响

›› 2013, Vol. ›› Issue (5): 133-139.

固醇调节元件结合蛋白基因的过表达对促进牛胎儿成纤维细胞脂肪形成的影响

徐丽, 郭立平, 朱淼, 高雪, 张路培, 高会江, 李俊雅, 许尚忠

中国农业科学院北京畜牧兽医研究所, 农业畜禽遗传资源与利用重点开放实验室, 北京 100193

修回日期:2013-04-19 出版日期:2013-05-20 发布日期:2013-05-27
通讯作者: 许尚忠,研究员,博士生导师,研究方向:肉牛遗传育种。E-mail:simmenta@vip.sina.com E-mail:simmenta@vip.sina.com
作者简介:徐丽(1986-),女,河北人,硕士生,研究方向:生物技术与动物遗传育种。
基金资助:
"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)"。

Effect of the Overexpression on Sterol Regulatory Element Binding Factor 1 Gene Increases Lipid Droplet Formation in Bovine Fetal Fibroblasts

XU Li, GUO Li-ping, ZHU Miao, GAO Xue, ZHANG Lu-pei, GAO Hui-jiang, LI Jun-ya, XU Shang-zhong

Key Laboratory of Farm Animal Genetic Resources and Utilization of Ministry of Agriculture, Institute of Animal Sciences, Chinese Academy of Agricultural Sciences, Beijing 100193, China

Revised:2013-04-19 Online:2013-05-20 Published:2013-05-27

摘要/Abstract

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48 h,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SREBP1 mRNA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。

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

Abstract: 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.

Key words: sterol regulatory element binding factor 1; overexpression; bovine fetal fibroblasts; lipid droplet formation; plasmid transfection

中图分类号:

S816.79

徐丽, 郭立平, 朱淼, 高雪, 张路培, 高会江, 李俊雅, 许尚忠. 固醇调节元件结合蛋白基因的过表达对促进牛胎儿成纤维细胞脂肪形成的影响[J]. , 2013, (5): 133-139.

XU Li, GUO Li-ping, ZHU Miao, GAO Xue, ZHANG Lu-pei, GAO Hui-jiang, LI Jun-ya, XU Shang-zhong. Effect of the Overexpression on Sterol Regulatory Element Binding Factor 1 Gene Increases Lipid Droplet Formation in Bovine Fetal Fibroblasts[J]. , 2013, (5): 133-139.

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