JAZF1基因抑制对3T3-L1脂肪细胞糖、脂代谢的影响  

Effects of JAZF1 gene inhibition on glucose and lipid metabolism in 3T3-L1 adipocytes and hepatocytes

作　　者：冉文侠[1] 李伶[1] 杨刚毅[1] 杜林[1] 罗小河[1] 贾彦军[1] 张亚丽[1] 罗妤[1] 

机构地区：[1]重庆医科大学检验医学院临床检验诊断学教育部重点实验室,重庆400016

出　　处：《中国老年学杂志》2013年第16期3894-3896,共3页Chinese Journal of Gerontology

基　　金：国家自然科学基金项目(30771037;81070640;30871199;30971388);教育部博士点基金(20105503110002);重庆市自然科学基金重点项目(cstc2012 jjB10022)

摘　　要：目的探讨JAZF1基因抑制对3T3-L1脂肪细胞糖、脂代谢相关基因的影响。方法构建JAZF1小发夹RNA(shRNA)表达载体并转染3T3-L1细胞,实时荧光定量PCR(RT-QPCR)和蛋白印迹法检测JAZF1 mRNA和蛋白水平的表达;氢三放射示踪法检测3T3-L1细胞糖摄取率;蛋白印记法检测糖、脂代谢相关基因蛋白水平;油红O染色检测脂肪细胞甘油三酯(TG)含量变化。结果成功构建JAZF1-shRNA;转染脂肪细胞48 h后,JAZF1 mRNA和蛋白水平明显低于对照组(P<0.05);氢3放射性示踪法显示转染组葡萄糖摄取率明显降低(P<0.05);PPAR-γ蛋白表达升高(P<0.05),激素敏感脂肪酶(HSL)、内脏脂肪素(Visfatin)、胰岛素诱导基因-2(Insig-2)蛋白表达降低(均P<0.05);油红O染色显示JAZFl转染组细胞内脂质积聚明显,比对照组升高约25%(P<0.05)。结论 JAZF1基因抑制可减少基础糖转运,增加脂质与胆固醇合成,减少脂质分解并减少相关脂肪细胞因子的表达。Objective To investigate the effects of juxtaposed with another zinc finger gene 1（JAZF1） suppression on glycolipids metabolism in 3T3-L1 adipocytes.Methods Short hairpin RNA（shRNA） targeting to JAZF1 was constructed and transfected into 3T3-L1 adipocytes.The mRNA and protein levels of JAZF1 were determined by RT-QPCR and Western blot.Glucose uptake was determined by 3H radioactive tracing;the expression of proteins related to glycolipids metabolism was measured by Western blot.Intracellular lipid accumulation was detected by oil red O staining method.Results The recombinant plasmid containing JAZF1 shRNA was constructed successfully.In JAZF1-shRNA transfected adipocytes,JAZF1 mRNA and protein levels were significantly（P0.05） lower than those of control.Glucose uptake was decreased significantly in transfected JAZF1-shRNA adipocytes（P0.05） after 48 h.The PPAR-γ protein level was increased significantly（P0.05）.The protein levels of HSL,visfatin and insulin induced gene（Insig）-2 were decreased compared to negative control（P0.05）.Oil red O staining showed intracellular lipid accumulation was increased significantly compared with that of negative control.Conclusions The suppression of JAZF1 in 3T3-L1 cells could increase lipid and cholesterol synthesis and improve lipid accumulation,reduce glucose transport and lipidolysis.

关 键 词：JAZF1 基因抑制 3T3-L1细胞 糖、脂代谢 

分 类 号：R333.6[医药卫生—人体生理学]