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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 investigate the effect of NYGGF4 gene silencing on the mitochondrial dynamics of 3T3-L1 mature adipocytes. Methods 3T3-L1 preadipocytes were used as tools to construct NYGGF4 gene silencing cell lines and control cell lines, respectively. Dexamethasone (DEX), 1-methyl-3-isobutylxanthine The insulin combination regimen induced the differentiation of precursor adipocytes into mature adipocytes. Mfn 1/2, Drp 1, Fis 1 mRNA and mitochondrial DNA (mtDNA) copy number of mature adipocytes were detected by real-time fluorescence quantitative PCR. Results The expression of Mfn1 / 2 and Drp1 in NYGGF4 silenced mature adipocytes was similar to that in control group. The expression of Fis1 in NYGGF4 gene silencing mature adipocytes was significantly higher than that in control group (P <0.05). Conclusion Silencing of NYGGF4 gene may up-regulate the expression of Fis1 gene in 3T3-L1 adipocytes, suggesting that silence of NYGGF4 gene may affect the mitochondrial dynamics of mature adipocytes by affecting mitochondrial division.