过表达Sirt2抑制猪前体脂肪细胞的分化

本文旨在利用过表达技术研究Sirt2在猪前体脂肪细胞分化中的作用.首先将Sirt2插入腺病毒穿梭载体pAdTrack-CMV,并与骨架载体pAdEasy-1在大肠杆菌BJ5183中同源重组,重组体用Lipofectamine2000包装转染HEK293细胞系,成功获得重组腺病毒vAd-Sirt2.用vAd-Sirt2感染猪前体脂肪细胞,48h后油红O染色法观察脂肪细胞分化情况,RT-PCR检测脂肪细胞分化标志基因PPARγ和aP2的表达.结果显示,过表达Sirt2促使细胞中脂滴减少,同时标志基因PPARγ、aP2mRNA水平显著降低,说明Sirt2抑制猪前体脂肪细胞分化,这为控制猪体脂沉积提供依据以及为人类肥胖和相关疾病的治疗和预防奠定基础.

Overexpression of Sirt2 Suppresses Porcine Preadipocyte Differentiation

The effect of Sirt2 on porcine preadipocyte differentiation was evaluated. ASirt2 overexpression plasmid was constructed with pAdTrack CMV, then linearized and shuttled into the BJ5183 bacteria strain harboring a supercoiled pAdEasy 1 backbone vector for recombination. The PacⅠ digested recombinant adenoviral DNA were transfected into HEK 293 cells by Lipofectamine 2000 to produce the vAd Sirt2 adenoviruse . Porcine preadipocytes were infected with vAd Sirt2 for 48 hours, Oil Red O staining and RT PCR for adipocyte differentiation marker genes (like PPARγand aP2 ) were performed. We found that the overexpression of Sirt2 significantly decreased the numbers of lipid droplets and the mRNA levels of adipocyte differentiation. The fact that Sirt2 to suppress porcine preadipocyte differentiation suggested that it might be a regulatory target to control the body fat deposition in pigs or to treat/prevent obesity related disorders in humans.