未知功能基因JAZF1过表达对3T3-L1脂肪细胞糖脂代谢的影响  

Effect of novel gene JAZF1 overexpression on glucose and lipid metabolism in 3T3-L1 adipocytes

作　　者：杨艳[1] 李伶[1] 杨刚毅[2] 孙滨[1] 卢春敏[1] 苗宗玉[1] 

机构地区：[1]重庆医科大学检验系临床生化教研室和教育部临床检验诊断学重点实验室,400016 [2]重庆医科大学附属第二医院内分泌科

出　　处：《中华内分泌代谢杂志》2010年第7期595-598,共4页Chinese Journal of Endocrinology and Metabolism

基　　金：国家自然科学基金项目（30771037,30871199,30971388）;重庆医科大学校级重点项目（XBZD200704）

摘　　要：目的 观察JAZF1基因（Juxtaposed with another zincfinger gene 1）过表达对3T3-L1脂肪细胞糖脂代谢相关基因的影响.方法 采用实时荧光定量PCR（RT-QPCR）法检测JAZF1 mRNA在健康C57BL/6J小鼠多种组织表达分布情况;构建JAZF1真核表达载体并瞬时转染3T3-L1细胞,RT-QPCR法检测JAZF1、糖脂代谢相关基因mRNA的表达;用Western印迹法测定各组细胞JAZF1蛋白水平;油红O染色比色法检测细胞内脂质积聚的变化.结果 转染48 h后,JAZF1转染组脂肪细胞中,JAZF1 mRNA及蛋白表达明显高于阴性对照和空载组,激素敏感脂肪酶（HSL）mRNA表达水平明显增加（P〈0.05）,脂肪酸合成酶（FAS）、乙酰辅酶A羧化酶（ACC）、类固醇调节元件结合蛋白1（SREBP1）mRNA相对表达量明显降低（均P〈0.01）,脂肪细胞甘油三酯酶（ATGL）、葡萄糖转运子1（GLUT1）、GLUT4 mRNA表达并无明显改变;油红O染色显示JAZF1转染组细胞内脂质积聚明显降低（P〈0.05）.结论 JAZFl在C57BL/6J小鼠多种组织均有表达,提示其可能在维持正常生理功能中起着一定作用.3T3-L1细胞过表达JAZF1可减少脂质合成、增加脂解,并可明显改善脂质积聚,可能是肥胖和糖尿病治疗的一个潜在靶点.Objective To observe the effects of JAZF1 （Juxtaposed with another zinc finger gene 1 ） overexpression on glucose and lipid metabolism in 3T3-L1 adipocytes. Methods The tissue distribution of JAZF1 in healthy C57BL/6J mice was detected by real-time quantitative PCR（ RT-QPCR）. Expression vector for JAZF1 gene was constructed and transfected into 3T3-L1 adipocytes. The mRNA levels of JAZF1, GLUT1, GLUT4, FAS, ACC, SREBP1, ATGL, and HSL implicated in glucose and lipid metabolism were determined by RT-QPCR; JAZF1 protein level was measured by Western blot. Intracelluar lipid accumulation were measured by oil red O staining method. Results In JAZF1-transfected adipocytes, JAZF1 mRNA and protein levels were significantly higher than control cells after 48 h. The mRNA level of HSL was increased significantly （P〈0. 05） in JAZF1 transfection group compared with negative control and empty vector group, and the expressions of FAS, ACC, SREBP1 mRNA were decreased significantly（all P〈0.01）. However, the mRNA levels of ATGL, GLUT1, GLUT4 were not changed. Intracelluar lipid accumulation was decreased significantly （P〈0.05 ） by oil red O staining and colorimetric in JAZF1 -transfected cells compared with negative control and empty vector group. Conclusions There was an extensive expression of JAZF1 in various tissues of C57BL/6J mice,indicating that JAZF1 might play a role in maintaining normal physiological function. These results show that overexpression of JAZF1 in 3T3-L1 cells can reduce lipid synthesis, increase lipolysis, and improve lipid accumulation. JAZF1 might provide a new potential therapeutic target for obesity and diabetes.

关 键 词：JAZF1 3T3-L1细胞 糖代谢 脂代谢 实时荧光定量PCR 脂质积聚 

分 类 号：R587.1[医药卫生—内分泌]