橄榄苦苷通过抑制PPARγ稳定糖脂代谢调控NAFLD的作用机制
Mechanism of Oleuropein on NAFLD by stabilizing glycolipid metabolism in NAFLD through inhibiting PPARγ
罗明珠 1王怡婷 1楚薇 1马琰岩 1王靖怡 1李晶哲 1刘长振1
作者信息
- 1. 中国中医科学院医学实验中心,北京 100700
- 折叠
摘要
目的:基于PPARγ信号通路探索橄榄苦苷对非酒精性脂肪肝病(NAFLD)细胞模型以及糖脂代谢的作用及可能机制.方法:采用油酸诱导AML12小鼠肝细胞构建NAFLD细胞模型.分成对照组、NAFLD组、GW9662组和橄榄苦苷组.用油红染色以及油红定量检测药物对NAFLD细胞模型脂质沉积的影响.基于表面等离子共振技术配体结合检测法和Luciferase体外转录激活实验,检测橄榄苦苷能否与PPARγ直接结合以及对其作用特性.采用qPCR法检测橄榄苦苷对PPARγ下游脂代谢相关靶基因的影响.采用流式细胞分析和生化分析检测橄榄苦苷对糖摄取的作用.采用3T3-L1前脂肪细胞研究橄榄苦苷对脂肪生成的影响.结果:与对照组比较,NAFLD组经油酸诱导后产生大量脂质沉积(P＜0.01),PPARγ及下游靶基因mRNA表达增加,如脂肪酸摄取和脂肪从头合成基因.与NAFLD组比较,GW9662组和橄榄苦苷组显著改善油酸诱导的脂肪堆积(P＜0.05).橄榄苦苷可以直接结合PPARγ并抑制其转录活性.橄榄苦苷降低PPARγ/α及其下游脂代谢相关靶基因mRNA表达水平(P＜0.05),如CD36、Fatp1、Fabp3、SCD1、Acc1和Fasn等.橄榄苦苷还可以在一定程度上促进AML12的糖摄取能力(P＜0.05),抑制3T3-L1细胞成脂分化(P＜0.05).结论:橄榄苦苷可以直接靶向PPARγ并抑制其转录活性,稳定糖脂代谢改善NAFLD.
Abstract
Objective:To investigate the effects and its potential mechanism and of oleuropein on non-alcoholic fatty liver disease(NAFLD)cell model and glucolipid metabolism via PPARγ signal pathway.Methods:Mouse hepatic AML12 cells induced by oleic acid were selected as NAFLD models.Cells were divided into control group,NAFLD group,positive control group and oleuropein group.The lipid accumulation of every group evaluated by oil red O staining.Binding and interaction with oleuropein and PPARγ were detected by ligand binding assay and luciferase transcriptional activity experiment in vitro.Lipid metabolic related downstream gene of PPARγ were determined by qPCR.Glucose intake level were detected by flow cytometry and biochemical analysis.Adipocyte differentiation of 3T3-L1 cells were evaluated.Results:Compared with control group,NAFLD group showed mounting lipid deposition(P＜0.01),PPARγ and its downstream gene mRNA increased significantly,including fatty acids uptake and fat de novo synthesis.Germpared with NAFLD group,the lipid droplets were reduced in positive control group and oleuropein group(P＜0.05).Oleuropein could indirect bind PPARγ and inhibit its transcriptional activity,decrease mRNA expression levels of PPARγ/α and its downstream genes,such as CD36,Fatpl,Fabp3,SCD1,Accl and Fasn.In addition,oleuropein promoted the glucose uptake to a certain extent(P＜0.05),and inhibit adipocyte differentiation of 3T3-L1 cells(P＜0.05).Conclusion:Oleuropein could stabilize glucolipid metabolism in NAFLD cell model via directly bind PPARγ and inhibit its transcriptional activity.
关键词
橄榄苦苷/非酒精性脂肪肝病/PPARγ/AML12小鼠肝细胞/3T3-L1前脂肪细胞Key words
Oleuropein/Non-alcoholic fatty liver disease(NAFLD)/PPARγ/AML12 cells/3T3-L1 cells引用本文复制引用
基金项目
北京市自然科学基金项目(7212187)
中国中医科学院科技创新工程项目(C12021A05021)
中央级公益性科研院所基本科研业务专项资金(JBGS2021004)
出版年
2024
NETL
NSTL
万方数据