敲减OGT对肝细胞脂肪合成的作用研究  

Effect of knockdown of O-Glcnac transferase on hepatocyte fat synthesis

作　　者：苗向霞 马澜婧[2] 曹嘉谊 王晨晨 聂勇战[2] 刘凯歌[1] Miao xiangxia;Ma Lanjing;Cao Jiayi;Wang Chenchen;Nie Yongzhan;Liu Kaige(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)

机构地区：[1]西安医学院第一附属医院消化内科,710077 [2]第四军医大学西京消化病医院肿瘤生物学国家重点实验室,西安710032

出　　处：《中华肝脏病杂志》2020年第2期147-151,共5页Chinese Journal of Hepatology

基　　金：陕西省科技计划社会发展攻关项目(2016SF-327);陕西省教育厅科研资助项目(11JK0708)。

摘　　要：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(SCD1)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.

关 键 词：脂肪肝 非酒精性 脂肪合成酶 O-连接的β-N-乙酰葡糖胺化 N-乙酰氨基葡萄糖转移酶 

分 类 号：R575[医药卫生—消化系统]