Cideb促进棕榈酸诱导的HepG2细胞脂质沉积  

Cideb promotes palmitic acid-induced lipid deposition in HepG2 cells

作　　者：杨林泉 佟斯羽 朱慧明 张会欣 王超[1] YANG Linquan;TONG Siyu;ZHU Huiming;ZHANG Huixin;WANG Chao(Key Laboratory of Metabolic Disease,Hebei General Hospital,Shijiazhuang 050051,China;Biochemistry Major at University of Wisconsin-Madison,Madison,WI 53706,USA;Chemical Engineering Institute,Shijiazhuang University,Shijiazhuang 050035,China)

机构地区：[1]河北省人民医院代谢病重点实验室,河北石家庄050051 [2]威斯康星大学麦迪逊分校,美国威斯康星州麦迪逊市53706 [3]石家庄学院化工学院,河北石家庄050035

出　　处：《中国病理生理杂志》2025年第3期509-517,共9页Chinese Journal of Pathophysiology

基　　金：国家自然科学基金面上项目(No.82170878);河北省卫健委科研计划项目(No.20190283)。

摘　　要：目的:探讨诱导细胞死亡DNA片段化因子α(DNA fragmentation factor alpha,DFFA)样效应因子B(cell death-inducing DFFA-like effector B,Cideb)促进棕榈酸(palmitic acid,PA)诱导的人肝母细胞瘤HepG2细胞脂质沉积的作用及机制。方法:将HepG2细胞分为对照组、模型组、沉默对照(siR-NC)组和Cideb沉默(siR-Cideb)组,采用PA诱导HepG2细胞建立脂质沉积细胞模型,利用特异性siRNA沉默Cideb的表达,检测细胞内脂质含量,油红O染色观察脂质沉积,RT-qPCR和Western blot法检测细胞中Cideb、固醇调节元件结合蛋白1c(sterol regulatory element-binding protein 1c,SREBP1c)、乙酰辅酶A羧化酶1(acetyl-CoA carboxylase 1,ACC1)、脂肪酸合成酶(fatty acid synthase,FAS)、硬脂酰辅酶A去饱和酶1(stearoyl-CoA desaturase 1,SCD1)和AMP活化蛋白激酶α(AMP-activated protein kinaseα,AMPKα)表达。通过转染Cideb真核表达质粒,观察过表达Cideb对HepG2细胞的脂质沉积影响。结果:与对照组比较,PA模型组出现脂质沉积,甘油三酯(triglyceride,TG)和总胆固醇(total cholesterol,TC)含量显著上升,Cideb、SREBP1c、ACC、FAS和SCD1表达水平升高,AMPKα表达水平降低(P<0.05);与siR-NC组比较,siR-Cideb能够抑制HepG2细胞脂质沉积,降低细胞内TG和TC含量,降低SREBP1c、ACC、FAS和SCD1表达水平,增加AMPKα表达(P<0.05);Cideb过表达能够增加HepG2细胞脂质沉积及TG和TC含量,增加SREBP1c、ACC、FAS和SCD1表达,降低AMPKα表达水平。结论:降低Cideb水平可通过下调SREBP1c/ACC/FAS/SCD1通路,上调AMPKα,从而抑制PA诱导的HepG2细胞脂质沉积。AIM:To investigate the role and mechanism of cell death-inducing DNA fragmentation factor al-pha(DFFA)-like effector B(Cideb)in promoting palmitic acid(PA)-induced lipid deposition in human hepatoblastoma HepG2 cells.METHODS:The HepG2 cells were divided into control group,model group,negative control silencing(siR-NC)group and Cideb silencing(siR-Cideb)group.The cells were treated with PA to establish a cellular model of lipid deposition.Specific siRNAs were used to silence the expression of Cideb,and the intracellular lipid content was mea-sured.Lipid deposition was observed by oil red O staining.RT-qPCR and Western blot were used to detect the expression levels of Cideb,sterol regulatory element-binding protein 1c(SREBP1c),acetyl-CoA carboxylase 1(ACC1),fatty acid synthase(FAS),stearoyl-CoA desaturase 1(SCD1)and AMP-activated protein kinaseα(AMPKα)in the cells.The ef-fects of Cideb overexpression on HepG2 cells were observed by transfection with a Cideb eukaryotic expression plasmid.RESULTS:Compared with control group,the cells in PA model group presented lipid deposition,significantly increased triglyceride(TG)and total cholesterol(TC)content,elevated expression levels of Cideb,SREBP1c,ACC,FAS and SCD1,and decreased expression of AMPKα(P<0.05).Compared with siR-NC group,the cells in siR-Cideb group pre-sented decreased lipid deposition,reduced intracellular TG and TC content,decreased SREBP1c,ACC,FAS and SCD1 expression levels,and increased AMPKαexpression(P<0.05).The overexpression of Cideb increased the lipid deposi-tion,TG and TC content,and the expression levels of SREBP1c,ACC,FAS and SCD1 in HepG2 cells,but decreased the expression of AMPKα(P<0.05).CONCLUSION:Reduction of Cideb alleviates PA-induced lipid deposition in HepG2 cells by down-regulating the SREBP1c/ACC/FAS/SCD1 pathway and up-regulating the AMPKαpathway.

关 键 词：Cideb蛋白 非酒精性脂肪性肝病 HEPG2细胞 脂质沉积 SREBP1c/ACC/FAS/SCD1通路 AMPKα通路 

分 类 号：R966[医药卫生—药理学] Q291[医药卫生—药学] R363.2[生物学—细胞生物学]