NYGGF4基因表达沉默对3T3-L1脂肪细胞线粒体动力学的影响

Effects of NYGGF4 gene silencing on mitochondrial dynamics in 3T3-L1 adipocytes

目的探讨NYGGF4基因表达沉默对3T3-L1成熟脂肪细胞线粒体动力学的影响。方法以3T3-L1前体脂肪细胞为工具,分别构建NYGGF4基因表达沉默细胞株和空载对照细胞株,通过地塞米松(DEX)、1-甲基-3-异丁基黄嘌呤(MIX)、胰岛素联合方案将前体脂肪细胞诱导分化为成熟脂肪细胞。采用实时荧光定量PCR检测成熟脂肪细胞中线粒体融合基因(Mfn)1/2、线粒体动态相关基因(Drp)1、线粒体分裂基因(Fis)1的mRNA表达水平和线粒体DNA(mtDNA)拷贝数。结果 NYGGF4基因表达沉默成熟脂肪细胞的Mfn1/2、Drp1表达水平与对照组相仿。NYGGF4基因表达沉默成熟脂肪细胞的Fis1的表达水平显著高于对照组(P<0.05)。结论在3T3-L1脂肪细胞中,NYGGF4基因表达沉默可显著上调Fis1基因的表达水平,提示NYGGF4基因表达沉默可能通过影响线粒体分裂从而影响成熟脂肪细胞线粒体动力学。

Objective To explore the effects of NYGGF4 gene silencing on mitochondrial dynamics in adipocytes.Methods With 3T3-L1 preadipocytes the blank expression vector（group A） and NYGGF4 gene silencing vector（group B） were constructed,which was induced to differentiate into mature adipocytes.The expressions of Mfn1/2,Drp1 and Fis1 mRNA and mitochondrial DNA copy number were detected by real-time fluorescent quantitative PCR.Results There was no significant difference in the levels of Mfn1/2 mRNA and Drp1 mRNA between two groups.Fis1 mRNA expression was remarkably higher in the adipocytes of group B than those of group A（P〈0.05）.Conclusion In the adipocytes with NYGGF4 gene silencing,the expression of Fis1 mRNA is up-regulated,indicating that mitochondrial dynamics of mature adipocytes is influenced by NYGGF4 gene silencing.