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目的 探讨敲减N-乙酰氨基葡萄糖转移酶(OGT)对肝细胞脂肪合成的影响. 方法 建立L02正常肝细胞脂肪变模型,蛋白免疫印迹法(Western blot)检测OGT及O-连接糖基化(O-GlcNAc)表达.建立L02细胞OGT敲减细胞系,油酸诱导后检测其脂质形成能力.实时荧光定量PCR (qRT-PCR)和Western blot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.结果 Western blot显示L02细胞脂肪变后OGT及O-GlcNAc表达均升高(P＜0.05).shOGT慢病毒感染L02细胞后,OGT mRNA相对表达水平明显下调(P＜0.01).油红O染色显示,L02 shOGT细胞内脂质减少,qRT-PCR显示系列脂肪合成酶:乙酰辅酶A羟化酶、脂肪酸合成酶和硬脂酰辅酶A去饱和酶mRNA相对表达水平均降低,差异具统计学意义(P值均＜0.05),蛋白水平与mRNA相对表达量一致.结论 敲减OGT后可通过降低O-GlcNAc水平抑制肝细胞脂肪合成.“,”Objective To investigate the effect of knockdown of O-GlcNAc transferase (OGT) on hepatocyte fat synthesis.Methods Liver cell line L02 were used to established the model of hepatic steatosis.The levels of OGT and O-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.Results Western blot showed that the expression of OGT and O-GlcNAc was increased in L02 cells after adipogenesis (P ＜ 0.05).After shOGT lentivirus infects L02 cells,OGT mRNA levels were down-regulated (P ＜ 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 (SCDl) were decreased,the difference was statistically significant (P ＜ 0.05),protein Expression is consistent with mRNA expression.Conclusion Knockdown of OGT can inhibit hepatocyte fat synthesis by reducing O-GlcNAc levels.