肝X受体激动剂直接激活肝细胞NgBR的表达  

Liver X receptor agonists directly up-regulate NgBR expression in hepatocytes

作　　者：贺晓雨 马佳玲 闫亚丽 陈元利 HE Xiaoyu;MA Jialing;YAN Yali;CHEN Yuanli(School of Food and Biological Engineering,Hefei University of Technology,Hefei 230601,China)

机构地区：[1]合肥工业大学食品与生物工程学院,安徽合肥230601

出　　处：《合肥工业大学学报（自然科学版）》2024年第6期784-789,817,共7页Journal of Hefei University of Technology：Natural Science

基　　金：国家自然科学基金资助项目(31770863)。

摘　　要：Nogo-B受体(Nogo-B receptor,NgBR)参与脂肪肝和胰岛素敏感性的形成,但是并不清楚肝X受体(liver X receptor,LXR)激动剂是否能够调控NgBR的表达。文章使用人工合成的LXR激动剂(T0901317和GW3965)分析其对肝源细胞系中NgBR表达的影响,构建正常或突变NgBR启动子,通过双荧光素酶报告基因系统检测LXR激动剂对启动子活性的影响;采用CRISPR-CAS9方法建立LXRα或LXRβ基因敲除的HepG2细胞系,通过Western Blot检测相关基因的表达变化;向ApoE-/-小鼠腹腔注射LXR激动剂T0901317,分析小鼠肝脏中NgBR的表达变化。结果发现,LXR激动剂能够通过激活LXR促进NgBR蛋白的表达,该诱导作用是以LXRE依赖的方式进行的,并且LXR的表达发挥着重要作用。在体内实验中,也证明了LXR激动剂T0901317上调NgBR蛋白表达。结果表明,NgBR是LXR的靶蛋白,LXR通过结合NgBR启动子LXRE序列促进其转录和翻译。Nogo-B receptor(NgBR)involves in the development of hepatic steatosis and insulin resistance.However,it is unclear whether liver X receptor(LXR)agonists can regulate NgBR expression in the liver.In this study,the effects of synthetic LXR agonists(T0901317 and GW3965)on NgBR expression in hepatic cell lines were analyzed.The normal and mutant NgBR promoters were constructed,and the effects of LXR agonists on promoter activity were detected by a dual luciferase reporter assay.Then,LXRαor LXRβknockout HepG2 cell lines were established according to the CRISPR-CAS9 method,and changes in expression of the relevant genes were determined by Western Blot.Finally,T0901317 was injected intraperitoneally into ApoE-/-mice to analyze the changes of NgBR expression in the liver of mice.The results revealed that LXR agonists were able to promote the expression of NgBR protein by activating LXR,which occurred in an LXRE-dependent manner,and LXR expression played an important role.In vivo experiments demonstrated that LXR agonist T0901317 up-regulated NgBR protein expression in the liver.The results suggest that NgBR is a target protein of LXR,and LXR promotes its transcription and translation by binding to the LXRE sequence in the promoter region of NgBR.

关 键 词：肝X受体(LXR) Nogo-B受体(NgBR) LXR激动剂 肝脏 

分 类 号：R543.1[医药卫生—心血管疾病]