论文部分内容阅读
【目的】观察NYGGF4(PID1)基因过表达对脂肪细胞线粒体代谢的影响。【方法】以3T3-L1前体脂肪细胞为研究对象,体外培养稳定转染NYGGF4(PID1)基因(NYGGF4-pcDNA3.1(+)/myc-His B)的3T3-L1前体脂肪细胞,以转染空载体[pcDNA3.1(+)/myc-His B]的3T3-L1前体脂肪细胞为对照,经1-甲基-3异丁基黄嘌呤(MIX)加地塞米松、胰岛素方案诱导分化为成熟脂肪细胞,采用Real-time PCR检测线粒体代谢酶指标:己糖激酶(Hexokinase,HKI)、乙酰辅酶A羧化酶(Acetyl-CoA,ACC)、柠檬酸合成酶(citrate synthase,CS)、肉毒碱棕榈酰转移酶1(Carnitine palmitoyl-transferase 1,CPT1)、细胞色素C(Cytochrome c,Cyc-c)基因表达水平。【结果】NYGGF4(PID1)过表达显著降低3T3-L1脂肪细胞己糖激酶、乙酰辅酶A羧化酶、柠檬酸合成酶、肉毒碱棕榈酰转移酶1、细胞色素C基因表达水平,差异有统计学意义。【结论】NYGGF4(PID1)基因过表达,下调3T3-L1脂肪细胞中定位于线粒体的代谢关键酶,提示NYGGF4(PID1)基因过表达能影响3T3-L1脂肪细胞的线粒体代谢。
【Objective】 To investigate the effect of NY1GFP4 over-expression on mitochondrial metabolism in adipocytes. 【Method】 The 3T3-L1 preadipocytes were cultured in vitro and stably transfected with 3T3-L1 preadipocytes stably transfected with the NYGGF4 gene (NYGGF4-pcDNA3.1 (+) / myc-His B) The 3T3-L1 preadipocytes transfected with empty vector [pcDNA3.1 (+) / myc-His B] served as controls and were induced by 1-methyl-3 isobutylxanthine (MIX) plus dexamethasone To differentiate into mature adipocytes, Real-time PCR was used to detect mitochondrial metabolic enzyme indexes: hexokinase (HKI), acetyl-CoA (ACC), citrate synthase (CS) , Carnitine palmitoyl-transferase 1 (CPT1), and Cytochrome c (Cyc-c) gene expression. 【Results】 Overexpression of NYGGF4 (PID1) significantly decreased the expression of hexokinase, acetyl-CoA carboxylase, citrate synthase, carnitine palmitoylase 1 and cytochrome C in 3T3-L1 adipocytes Statistical significance. 【Conclusion】 Overexpression of NYGGF4 (PID1) gene down-regulates the mitochondrial metabolic key enzyme in 3T3-L1 adipocytes, suggesting that overexpression of NYGGF4 (PID1) gene may affect mitochondrial metabolism in 3T3-L1 adipocytes.