·糖尿病基础研究·

　　【摘要】 目的 探讨锌α2糖蛋白(ZAG)对脂肪细胞线粒体生物合成相关因子mRNA和蛋白表达的影响。 方法 构建重组小鼠ZAG真核表达质粒pcDNA3.1(-)-mZAG，经脂质体转染3T3-L1细胞，24 h后提取RNA和蛋白质。RT-PCR法及Western Blot法检测过氧化物酶增殖型受体γ辅助活化因子1α(PGC-1α)、核呼吸因子1(NRF-1)、核呼吸因子2(NRF-2)、线粒体转录因子A(mtTFA) mRNA和蛋白的表达。 结果 与未转染组和pcDNA3.1(-)组比较，pcDNA3.1(-)-mZAG组3T3-L1细胞PGC-1α、NRF-1、NRF-2、mtTFA的mRNA和蛋白表达均上调(P<0.01)。 结论 ZAG对脂肪细胞线粒体生物合成相关因子PGC-1α、NRF-1，NRF-2、mtTFA的mRNA和蛋白表达均有促进作用。

　　【关键词】 锌α2糖蛋白;脂肪细胞;线粒体生物合成

Effect of zinc-α2-glycoprotein on the expression of relevant factors of mitochondria biogenesis in adipocytes XIAO Xin-hua, ZHANG Huan-li, RAN Li, et al. Department of Endocrinology, The First Affiliated Hospital, University of South China, Hengyang 421001, China

Corresponding author: LIU Jiang-hua, E-mail: jianghua990@hotmail.com

　　【Abstract】Objective To investigate the effect of zinc-α2-glycoprotein (ZAG) on the expression of relevant factors of mitochondria biogenesis in adipocytes. Methods The total RNA was extracted from mouse liver cells and the sequence of ZAG was amplified with RT-PCR. The PCR product was cloned into the prokaryotic expression vector pcDNA3.1(-) after enzyme by XbaІ and HindШ. After identification by double restriction enzyme digestion and DNA sequencing, the recombinant eukaryotic expression plasmid pcDNA3.1(-)-mZAG was transfected into 3T3-L1 cells by lipofectamine 2000. The stable transfected 3T3-L1 cell line was established after selection with G418. After 24 hours of transfection, the mRNA and protein expressions of PGC-1α, NRF-1, NRF-2, and mtTFA were detected by RT-PCR and Western blot respectively. Results The pcDNA3.1(-)-mZAG plasmid was constructed and transfected into the 3T3-L1 cells successfully. Compared with the untransfected group and pcDNA3.1(-) transfected group, the level of mRNA and protein expressions of PGC-1α, NRF-1, NRF-2, and mtTFA were significantly up-regulated in the pcDNA3.1(-)-mZAG transfected group (P<0.01). Conclusion The recombinant eukaryotic expression vector pcDNA3.1(-)-mZAG and 3T3-L1 cell line stable over-expression of ZAG were established. ZAG promotes mRNA and protein expressions of mitochondria biogenesis genes.

　　【Key words】 Zinc-α2-glycoprotein; Adipocyte; Mitochondria biogenesis