Miao X X, Ma L J, Cao J Y, Wang C C, Nie Y Z, Liu K G
Department of Digestive Medicine, First Affiliated Hospital of Xi'an Medical College, Xi'an 710077.
Xijing Hospital of Digestive Disease & State Key of Laboratory of Cancer Biology, Fourth Military Medical University, Xi'an 710032.
Zhonghua Gan Zang Bing Za Zhi. 2020 Feb 20;28(2):147-151. doi: 10.3760/cma.j.issn.1007-3418.2020.02.009.
To investigate the effect of knockdown of -GlcNAc transferase (OGT) on hepatocyte fat synthesis. Liver cell line L02 were used to established the model of hepatic steatosis. The levels of OGT and -GlcNAc protein were detected by Western blot. The OGT knockdown cell line of L02 cells was established, and its lipid formation ability was detected after induction of oleic acid (OA). Real-time quantitative PCR (qRT-PCR) and Western blot were used to detect mRNA and protein expression of enzymes related to fat synthesis. An independent sample t test was used. Western blot showed that the expression of OGT and -GlcNAc was increased in L02 cells after adipogenesis ( < 0.05). After shOGT lentivirus infects L02 cells, OGT mRNA levels were down-regulated ( < 0.01). Oil red O staining showed that the lipid in L02 shOGT cells decreased, qRT-PCR showed that the mRNA expressions of fat synthase (ACC1), (FASN) and (SCD1) were decreased, the difference was statistically significant ( < 0.05), protein Expression is consistent with mRNA expression. Knockdown of OGT can inhibit hepatocyte fat synthesis by reducing -GlcNAc levels.
为研究敲低O-连接的N-乙酰葡糖胺转移酶(OGT)对肝细胞脂肪合成的影响。采用肝细胞系L02建立肝脂肪变性模型。通过蛋白质免疫印迹法检测OGT和O-连接的N-乙酰葡糖胺蛋白水平。建立L02细胞的OGT敲低细胞系,诱导油酸(OA)后检测其脂质形成能力。采用实时定量聚合酶链反应(qRT-PCR)和蛋白质免疫印迹法检测脂肪合成相关酶的mRNA和蛋白表达。采用独立样本t检验。蛋白质免疫印迹法显示,成脂后L02细胞中OGT和O-连接的N-乙酰葡糖胺的表达增加(P<0.05)。慢病毒载体shOGT感染L02细胞后,OGT mRNA水平下调(P<0.01)。油红O染色显示,L02 shOGT细胞中的脂质减少,qRT-PCR显示,脂肪合成酶(ACC1)、(FASN)和(SCD1)的mRNA表达降低,差异具有统计学意义(P<0.05),蛋白表达与mRNA表达一致。敲低OGT可通过降低O-连接的N-乙酰葡糖胺水平抑制肝细胞脂肪合成。