siRNA特异性抑制犊牛原代肝细胞SREBP-1c基因的表达  

Specific inhibition effect of siRNA on SREBP-1c gene expression in calf primary hepatocyte

作　　者：邓清华[1] 付世新[1,2] 刘国文[1] 刘磊[1] 王建国[1] 白鸽[1] 朱晓岩[1] 李慧萍[3] 李小兵[1] 王哲[1] 

机构地区：[1]吉林大学畜牧兽医学院,吉林长春130062 [2]黑龙江八一农垦大学动物科技学院,黑龙江大庆163319 [3]吉林大学人兽共患病研究所,吉林长春130062

出　　处：《中国兽医学报》2011年第7期1050-1053,1058,共5页Chinese Journal of Veterinary Science

基　　金：国家自然科学基金资助项目(30972224;30871897)

摘　　要：固醇调节元件结合蛋白-1c(SREBP-1c)是脂肪合成基因的重要转录调节因子,又称脂肪细胞决定和分化因子1。SREBP-1c主要在动物肝脏和脂肪细胞中表达,是脂肪代谢中重要的核转录因子,它可以通过调节脂肪代谢相关酶基因的表达来调控动物体内的脂肪合成。本试验设计合成了SREBP-1c的siRNA,通过脂质体转染将siRNA转染进犊牛原代肝细胞,用实时荧光相对定量PCR检测siRNA对SREBP-1c基因表达量的影响。结果显示:siRNA通过脂质体成功转入犊牛原代肝细胞;与对照组相比,一组转染细胞在mRNA水平检测到较低的SREBP-1c基因表达量。这表明siRNA可以稳定地在犊牛原代肝细胞中表达,并且对靶基因呈现出抑制作用。该研究为在基因水平防治奶牛肝脂沉积提供了基础。Sterol regulatory element binding protein-1c （SREBP-1c） is; an important lipogenic gene transcription factor,also known as adipocyte determination and differentiation factor 1 （adipocyte determination and differentiation factor 1）. SREBP-1c,mainly found in liver and fat cells,the expression of which is an important nuclear transcription factor,can be related to fat metabolism by regulating the expression of genes regulating fat synthesis in animals. In the present study,the specific siRNA of SREBP lc was designed and synthesized,and then was transfected into pri mary liver cells of a calf by liposome. Real-time fluorescence relative quantitative PCR （RT-PCR） was employed to detecte the expression of SREBP-1c gene expression. The results showed that, siRNA could enter into the hepato cytes and the mRNA level of SREBP-1c of transfected cells was lower than that of the control group,indicating that siRNA could be stabilized in the primary liver cells of cows and displayed a certain extent of inhibition effect on the target genes. The research lay a foundation of the possibility of gene therapy for fatty deposits in cows.

关 键 词：犊牛原代肝细胞 SIRNA RNA干扰 固醇调节元件结合蛋白-1C 

分 类 号：S856.5[农业科学—临床兽医学]