脂联素siRNA对3T3-L1细胞基础葡萄糖转运的影响(英文)  

The Effects of Adiponectin siRNA on the Glucose Transport in 3T3-L1 Adipocytes

作　　者：孟凡萍[1] 郝坡[1] 王长本[1] 李良琼[1] 

机构地区：[1]重庆三峡中心医院临检科,重庆万州404000

出　　处：《现代生物医学进展》2011年第23期4412-4416,共5页Progress in Modern Biomedicine

基　　金：supported by a grant of General Program of Health Service of Chongqing(2008-2-387,2011-2-414);a grant of Science and Technique Foundation of Wanzhou,Chongqing(20084009,200903006)~~

摘　　要：目的:构建携带小鼠脂联素(Acrp30)siRNA腺病毒载体,并检测其对小鼠脂肪细胞Acrp30表达以及对3T3-L1脂肪细胞基础葡萄糖转运的影响。方法:设计并化学合成小鼠脂肪细胞Acrp30 siRNA片断,将其亚克隆入AdEaxy XL腺病毒载体系统,在293细胞内包装扩增为重组腺病毒。用此重组腺病毒感染3T3-L1脂肪细胞,用RT-PCR和ELISA检测其Acrp30 mRNA和蛋白表达。采用2 Deoxy-[3H]D-glucose掺入法测定脂肪细胞葡萄糖转运。结果:设计并构建了小鼠Acrp30基因特异性siRNA腺病毒载体,该载体感染脂肪细胞后,能显著抑制Acrp30 mRNA和蛋白表达,影响3T3-L1脂肪细胞基础葡萄糖的转运,与对照组相比,差异有显著性意义(P<0.05)。结论:构建的Acrp30基因特异性siRNA腺病毒载体能有效的抑制脂联素在3T3-L1脂肪细胞中的表达,从而影响3T3-L1脂肪细胞基础葡萄糖转运。Objective： To construct the RNA interference eukaryotic expression vector specific for Adiponectin （Acrp30） gene, and to investigate its effect on the Acrp30 expression and glucose transport in 3T3-L1 adipocytes. Methods： Mice Acrp30 siRNA fragment were designed, synthesized and cloned into the adenovirus vector. 3T3-L1 cells were infected with the two recombinant adenovirus- es respectively, The mRNA expression and protein levels of Acrp30 in these cells were evaluated by RT-PCR and ELISA. And Glucose transport is measured by 2-Deoxy- [3HI-D-glucose incorporation method. Results： The recombinant adenovirus were successfully constructed. They can remarkably down-regulate the expression of Acrp30 at both the mRNA and protein levels in 3T3-L1 cells, and decrease the glucose transport in 3T3-L1 adipocytes. （P〈0.05）. Conclusion： The siRNA expression vectors can effectively inhibited the expression of Acrp30 in 3T3-L1 adipocytes, and decrease the glucose transport in them.

关 键 词：RNA干扰腺病毒载体 脂联素 葡萄糖转运 

分 类 号：R589.2[医药卫生—内分泌]