NYGGF4基因过表达对脂肪细胞线粒体的影响  

Effects of NYGGF4 overexpression on mitochondria in adipocyte

作　　者：徐广峰[1] 赵亚萍[1] 陈小慧[2] 高春林[2] 王建国[1] 郭锡熔[2] 

机构地区：[1]中国人民解放军第82医院检验科,江苏淮安223001 [2]南京医科大学附属南京妇幼保健院,南京210004

出　　处：《临床检验杂志》2011年第4期276-278,共3页Chinese Journal of Clinical Laboratory Science

基　　金：国家自然科学基金(30571978;30973231);江苏省医学领军人才项目(LJ200624)

摘　　要：目的观察NYGGF4基因过表达对脂肪细胞线粒体的影响。方法构建NYGGF4基因表达载体,转染3T3-L1前体脂肪细胞,G418筛选表达NYGGF4细胞株后,用RT-PCR及western blot技术验证,建立NYGGF4稳定过表达细胞株。用透射电镜观察NYGGF4基因过表达对脂肪细胞线粒体形态的影响,实时荧光定量PCR检测NYGGF4基因过表达及对照空载脂肪细胞中核转录因子PGC1α、NRF1、mtTFA mRNA含量。结果 RT-PCR及western blot均提示NYGGF4稳定过表达细胞株建立成功。NYGGF4基因过表达可以导致脂肪细胞线粒体形态异常,显著上调脂肪细胞PGC1αmRNA表达,对NRF1、mtTFA mRNA水平无明显影响。结论 NYGGF4基因过表达可以导致脂肪细胞线粒体损伤。Objective To observe the effects of NYGGF4 overexpression on mitochondria in 3T3-L1 adipocyte.Methods The coding sequence of human NYGGF4 gene was inserted into the pcDNA3.1Myc/His B vector to become recombinant expression plasmid NYGGF4-pcDNA3.1/myc-His B which was subsequently transferred into 3T3-L1 pre-adipocyte.The transfected 3T3-L1 cell expressing NYGGF4 was selected with neomycin（G418）,and identified by RT-PCR and western blot.The effect of NYGGF4 overexpression on mitochondrial morphology in transfected 3T3-L1 was evaluated by transmission electron microscope.The levels of peroxisome proliferator-actived receptor co-activator 1α（PGC1α）,nuclear respiratory factor 1（NRF1） and mitochondrial transcription factor A（mtTFA） mRNA in the transfected 3T3-L1 and control cells were detected by real-time quantitative RT-PCR.Results RT-PCR and western blot demonstrated that the 3T3-L1 strain stably overexpressing NYGGF4 was established successfully.The overexpression of NYGGF4 may lead to abnormal mitochondrial morphology and increased PGC-1α mRNA level in adipocytes but no increased NRF1 and mtTFA mRNA was found.Conclusions The overexpression of NYGGF4 may damage mitochondia in adipocytes.

关 键 词：NYGGF4基因 脂肪细胞 线粒体 

分 类 号：R723.14[医药卫生—儿科]