利拉鲁肽通过Sirt1/AMPK信号通路改善肝细胞脂肪变性机制研究

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中文摘要：目的探讨利拉鲁肽(Lira)干预棕榈酸(PA)诱导的人肝癌细胞对沉默信息调节因子1(Sirt1)/腺苷酸活化蛋白激酶(AMPK)/固醇调节元件结合蛋白(SREBP1c)信号通路的影响。方法取HepG2细胞，分为5组，以PA干预诱导肝细胞脂肪变制备模型，再分别以PA/Lira、PA/Sirt1或PA/Sirt1/Lira干预组。采用油红O染色观察细胞脂滴，采用试剂盒测定肝细胞培养上清液烟酰胺腺嘌呤二核苷酸氧化态/还原态比值(NAD+/NADH)、丙氨酸氨基转移酶(ALT)、天冬氨酸氨基转移酶(AST)和肝细胞甘油三酯(TG)含量，采用RT-qPCR法检测肝激酶B1(LKB1)、游离脂肪酸(FFA)、乙酰辅酶A羧化酶(ACC)和甘油三酯脂肪酶(ATGL)mRNA水平，采用蛋白质印迹法检测细胞p-AMPK、p-Sirt1和p-SREBP1c蛋白表达。结果 PA处理组肝细胞内见大量脂滴，PA/Sirt1组脂滴更大，而PA/Lira处理组和PA/Sirt1/Lira处理组细胞脂滴数量显著减少，提示模型制备成功并显示Lira的防治作用;PA组细胞FFA和ACC基因水平显著高于对照组(P均＜0。01)，PA/Lira处理组LKB1和ATGL基因水平显著高于PA处理组(P均＜0。001)，而FFA和ACC基因水平显著低于PA组(P均＜0。01)，PA/Sirt1/Lira组LKB1和ATGL基因水平显著高于(P均＜0。01)，而FFA基因水平显著低于PA/Sirt1处理组(P＜0。05);与对照组比，PA组细胞p-AMPK和p-Sirt1蛋白表达下调，而SREBP1c表达上调;与PA组比，PA/Lira组p-AMPK蛋白表达上调而SREBP1c蛋白表达下调;与PA/Sirt1组比，PA/Sirt1/Lira组细胞p-AMPK表达上调。结论 Lira改善HepG2细胞脂肪蓄积可能是通过直接上调Sirt1/AMPK信号通路或部分激活LKB1升高AMPK蛋白表达，抑制SREBP1c蛋白表达，降低了脂质合成分子水平。

外文标题：Liraglutide ameliorates hepatocyte steatosis by regulation Sirt1/AMPK pathway in vitro

外文摘要：Objective This experiment was conducted to explore effect of liraglutide(Lira)on steatosis in HepG2 cells in vitro.Methods The HepG2 cells were divided into five groups,cell steatosis model were induced by 0.25 mmol/L palmitic acid(PA)incubation,and then intervened by silent information regulator 1(Sirt1)inhibitor,Lira or Sirt1 and Lira combination.Oil red O was stained for lipid droplets,and nicotinamide adenine dinucleotide oxidation/reduced state ratio(NAD+/NADH),ALT,AST and triglyceride(TG)contents were determined.Liver kinase B1(LKB1),free fatty acid(FFA),acetyl-CoA carboxylase(ACC)and adiposetriglyceride lipase(ATGL)mRNA loads were detected by RT-qPCR,and p-adenosine monophosphate-activated protein kinase(AMPK)/sterol regulatory element binding protein 1c(SREBP1c)and p-Sirt1 expression were detected by Western blot.Results Intrahepatocellular lipid droplets were clearly observed in PA-intervened HepG2 cells,which were even more severe in PA/Sirt1-intervened cells,while the lipid droplets obviously reduced in PA/Lira-and PA/Sirt1/Lira-intervened cells,suggesting the model establishment successful and protective effect of Lira;FFA and ACC mRNA loads in PA-intervened cells increased greatly compared to that in control cells(P＜0.01),LKB1 and ATGL mRNA loads in PA/Lira-treated cells increased greatly compared to that in PA-treated cells(P＜0.001),while FFA and ACC mRNA loads decreased greatly as compared to that in PA-treated cells(P＜0.01);LKB1 and ATGL mRNA loads in PA/Sirt1/Lira-treated cells were much higher(P＜0.01),while FFA mRNA loads was much lower than in PA/Sirt1-treated cells(P＜0.05);p-AMPK and p-Sirt1 protein expression in PA-intervened cells were down-regulated,while SREBP1c expression was up-regulated as compared to that in control cells;p-AMPK expression in PA/Lira-treated cells was up-regulated,while SREBP1c expression was down-regulated compared to that in PA-treated cells;p-AMPK expression in PA/Sirt1/Lira-treated cells was up-regulated compared to that in PA/Sirt1-treated cells.Conclusion Lira could attenuate intracellular fat accumulation in HepG2 cells in vitro,which might be ascribed to direct up-regulation of SIRT1/AMPK signaling pathway or to partially activation of LKB1 leading to increased expression of AMPK protein and inhibition of SREBP1c expression.

外文关键词：

Hep G2 CellsLiraglutideSilent information regulator 1Adenosine monophosphate-activated protein kinaseSteatosisIn vitro

作者：

路瑶、焦谊、古丽阿依木

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作者单位：

830000 乌鲁木齐市新疆医科大学第二附属医院内分泌科

830000 乌鲁木齐市新疆医科大学基础学院生物化学与分子生物学教研室

关键词：

HepG2细胞利拉鲁肽沉默信息调节因子1 腺苷酸活化蛋白激酶脂肪变体外

基金：

新疆维吾尔自治区自然科学基金资助项目省部共建中亚高发病成因与防治国家重点实验室基金资助开放课题新疆维吾尔自治区创新团队基金资助项目

项目编号：

2020D01C183SKL-HIDCA-2022-DX62022D14009

出版年：

2024

DOI：

10.3969/j.issn.1672-5069.2024.05.007

实用肝脏病杂志

中华医学会安徽分会

实用肝脏病杂志

CSTPCD

影响因子：1.362

ISSN：1672-5069

年,卷(期)：2024.27(5)