NYGGF4基因过表达对脂肪细胞线粒体的影响
Effects of NYGGF4 overexpression on mitochondria in adipocytes
投稿时间:2010-04-29  修订日期:2010-10-18
DOI:
中文关键词:  NYGGF4基因  脂肪细胞  线粒体
英文关键词:NYGGF4 gene  adipocyte  mitochondria
基金项目:国家自然科学基金(30571978,30973231);江苏省医学领军人才项目(LJ200624)
作者单位E-mail
徐广峰 江苏淮安解放军第82医院检验科 123248978@qq.com 
摘要点击次数: 2428
全文下载次数: 15
中文摘要:
      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,采用RT-PCR及westernblot技术进行验证,建立NYGGF4稳定过表达细胞株。采用透射电镜观察NYGGF4基因过表达对脂肪细胞线粒体形态的影响,荧光定量RT-PCR技术检测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(1)RT-PCR及westernblot检测均提示NYGGF4稳定过表达细胞株建立成功;(2)NYGGF4基因过表达可以导致脂肪细胞线粒体形态异常;(3)NYGGF4基因过表达可以显著上调脂肪细胞核转录因子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。
英文摘要:
      【Abstract】 Objective To observe the effects of NYGGF4 overexpression on mitochondria in 3T3-L1 adipocytes. Methods The coding sequence of human NYGGF4 was subcloned into the pcDNA3.1Myc/His (B) vector. Expression vectors carrying the NYGGF4 coding sequence or empty vectors were transfected into 3T3-L1 preadipocytes. Neomycin (G418) was added to the medium to select for transfected cells. The cells expressing human NYGGF4 was identified by RT-PCR and Westernblot. The mitochondrial morphology was observed on a transmission electron microscope. levels of PGC1α、NRF1、mtTFA mRNA expression were evaluated by real-time quantitative RT-PCR. Results (1) The 3T3-L1 cell-lines stably transfected with NYGGF4 gene has been established successfully. (2) NYGGF4 overexpression led to abnormal mitochondrial morphology in adipocytes; (3) NYGGF4 overexpression increased PGC-1α mRNA expression in adipocytes, but not affected NRF1, mtTFA mRNA levels. Conclusions NYGGF4 overexpression could impair mitochondia in adipocytes.
  查看/发表评论  下载PDF阅读器
关闭