NYGGF4基因表达沉默对3T3-L1脂肪细胞线粒体代谢的影响

Effect of NYGGF4 Gene Silencing on Mitochondrial Metabolism in 3T3-L1 Adipocytes

目的探讨NYGGF4(又称PID1)基因沉默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基因沉默载体(pGPU6/GFP/Neo-shRNA,NYGGF4-RNAi)的3T3-L1前体脂肪细胞,以转染空载体(pG-PU6/GFP/Neo-shRNA,NC shRNA)的3T3-L1前体脂肪细胞作为对照,经1-甲基-3异丁基黄嘌呤加地塞米松、胰岛素方案诱导分化为成熟脂肪细胞,采用实时定量RCR技术检测成熟脂肪细胞中线粒体代谢酶指标: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。结果 NYGGF4表达沉默与NYGGF4基因过表达相比,可以部分逆转3T3-L1脂肪细胞中己糖激酶、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,差异均有统计学意义。结论NYGGF4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基因的动态平衡对维持3T3-L1脂肪细胞的线粒体代谢具有重要作用。

Objective To explore the effect of knockdown NYGGF4 （ PID1 ） gene on mitochondrial metabolism in 3T3 - L1 adipocytes. Methods With 3T3 -L1 preadipocytes as a subject, which transfected with a NYGGF4 expression vector, an empty expression vector and NYGGF4 gene silencing vector and then the preadipoertes were induced to differentiate into adipocvtes using MIX, DXM and insulin. The ex- pression levels of metabolic enzymes in mitochondria hexokinase （HKI）,acetyl -CoA（ACC） ,citrate synthase（CS）, camitine palmitoyl - transferasel （cirri） ,cytochrome C （Cyc -C） were detected by real - time PCR. Results Knockdown NYGGF4 could reverse the level of metabolic enzymes in mitochondria HKI, ACC, CS, cirri, Cyc - c in NYGGF4 - overexperssion adipocytes. Conclusions In adipocytes with NYGGF4 gene overexpression, mitochondria metabolic enzymes mRNA is reversed in NYGGF4 gene silencing. It can implicate that mito- chondrial metabolism in 3T3 - L1 adipocytes is influenced by NYGGF4 gene.