穿心莲内酯拮抗雌激素相关受体α活性影响小肠脂质分泌的作用机制  

Andrographolide Inhibits Intestinal Lipid Secretion via Antagonizing Estrogen-Related ReceptorαActivity

作　　者：李利[1,2] 高军 王紫辉 张敏仪 吴尚英 姚冬生 管敏[2] LI Li;GAO Jun;WANG Zihui;ZHANG Minyi;WU Shangying;YAO Dongsheng;GUAN Min(Institute of Biomedicine,Jinan University,Guangzhou 510632,China;Shenzhen Institute of Advanced Technology,Chinese Academy of Sciences,Shenzhen 518055,China;University of Chinese Academy of Sciences,Beijing 100049,China)

机构地区：[1]暨南大学生物医药研究院,广州510632 [2]中国科学院深圳先进技术研究院,深圳518055 [3]中国科学院大学,北京100049

出　　处：《中国细胞生物学学报》2023年第7期1047-1057,共11页Chinese Journal of Cell Biology

基　　金：国家重点研发计划(批准号:2018YFA0703100);国家自然科学基金(批准号:82072493、82000739、81570532);广东省基础与应用基础研究基金(批准号:2022A1515010528、2019A1515110128);深圳市科技研究资助计划(批准号:JCYJ20210324101800002)资助的课题。

摘　　要：该文旨在研究雌激素相关受体α(estrogen-related receptorα,ERRα或ESRRA)拮抗剂穿心莲内酯(andrographolide,AP)对小肠脂质分泌功能的影响及其作用机制。采用荧光素酶报告基因系统探究AP对ERRα及其共激活因子过氧化物酶体增殖物激活受体共激活因子1α(peroxisome proliferator-activated receptor coactivator 1α,PGC1α)转录活性的影响。运用启动子报告基因系统、实时荧光定量PCR和Western blot方法研究AP对ERRα下游靶基因载脂蛋白B(apolipoprotein B,Apob)和微粒体甘油三酯转移蛋白(microsomal triglyceride transfer protein,Mttp)表达的抑制作用。通过脂滴荧光染色和甘油三酯(triglyceride,TG)定量实验分别检测AP处理后胞内的脂滴含量和胞内外TG含量。餐后甘油三酯应答实验、小肠油红O染色和TG定量分析研究AP对小鼠小肠脂质分泌的影响。结果显示,AP下调ERRα/PGC1α的转录活性,并影响ERRα/PGC1α对Apob和Mttp启动子的转录调控作用;AP下调ERRα下游靶基因Apob和Mttp的表达;AP处理后胞内脂滴含量增多,胞外TG含量减少;AP抑制小肠中脂质的分泌。结果证明,AP作为ERRα拮抗剂,下调ERRα/PGC1α介导的脂质分泌关键基因的转录,抑制小肠脂质的分泌,并降低小鼠血清中TG水平。The aim of this study is to investigate the effect and mechanism of AP(andrographolide),an ERRα(estrogen-related receptorα,also called ESRRA)inverse agonist,on lipid secretion of small intestine.Luciferase reporter assay was employed to identify whether AP decreases transcriptional activity of ERRαand its coactivitor PGC1α(peroxisome proliferator-activated receptor coactivator 1α).To determine if AP suppresses the promoter activity and gene expression of ERRαtarget genes Apob(apolipoprotein B)and Mttp(microsomal triglyceride transfer protein),promoter reporter activity,real-time quantitative PCR and Western blot were used.The intracellular lipid droplet content and TG content were determined by fluorescent staining of lipid droplets and TG(triglyceride)quantification experiments,respectively.Postprandial triglyceride response experiments,oil red O staining of small intestine and TG content determination were performed to study the effect of AP on lipid secretion in the small intestine of mice.The results showed that AP decreased promoter expression and mRNA levels of Apob and Mttp via interfering with transcriptional activity of ERRα/PGC1α.Furthermore,AP increased intracellular lipid droplets,reduced extracellular TG content and blood TG level indicating reduction of lipids secretion in Caco-2 and small intestine.This finding demonstrated that AP,as an ERRαinverse agonist,inhibited intestinal lipid secretion and decreased serum TG levels through downregulating the transcriptional ex­pression of Apob and Mttp.

关 键 词：穿心莲内酯 雌激素相关受体α 转录活性 脂质分泌 

分 类 号：R285[医药卫生—中药学]