脂联素siRNA对3T3-L1细胞基础葡萄糖转运的影响(英文)

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。

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 adenoviruses 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- 3H]-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.