脂联素siRNA对3T3-L1细胞基础葡萄糖转运的影响

Effects of adiponectin siRNA on the glucose transport in 3T3-L1 adipocytes

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片段,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,差异显著(P<0.05)。结论:构建的Acrp30基因特异性siRNA腺病毒载体能有效地抑制脂联素在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。

AIM： To construct the adenovirus vector with adiponectin （Acrp30） siRNA, and to observe its effect on the Acrp30 expression and glucose transport in 3T3 -L1 adipocytes. METHODS： Mouse Acrp30 siRNA fragment was 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. Glucose transport was measured by 2 - Deoxy - [ ^3H ] - D - glucose incorporation method. RESULTS： The recombinant adenoviruses were successfully constructed. They remarkably downregulated the expression of Acrp30 at both mRNA and protein levels in 3T3 -L1 cells, and decreased the glucose transport in 3T3 -L1 adipocytes （P 〈0. 05）. CONCLUSION： The siRNA expression vectors effectively inhibit the expression of Acrp30 in 3T3 - L1 adipocytes, and decrease the glucose transport.