脂联素siRNA对3T3-L1细胞基础葡萄糖转运的影响

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2Deoxy-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5C（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。

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

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.