角鲨烯对HepG2细胞LDLR表达的影响及机制初探  

Effects of squalene on expression of LDLR in HepG2 cells and its mechanism

作　　者：李婷[1] 赵丽萍 余超超 田男[1] 范春雷[1] LI Ting;ZHAO Li-ping;YU Chao-chao;TIAN Nan;FAN Chun-lei(College of Life Science,Zhejiang Chinese Medicical University,Hangzhou 310053,China)

机构地区：[1]浙江中医药大学生命科学学院,浙江杭州310053

出　　处：《中国药理学通报》2018年第7期1020-1024,共5页Chinese Pharmacological Bulletin

基　　金：浙江省自然科学基金资助项目(No LY14H290007)

摘　　要：目的研究角鲨烯对HepG2细胞低密度脂蛋白受体(LDLR)表达的影响,并探讨其降胆固醇的机制。方法MTT法检测不同浓度角鲨烯对HepG2细胞增殖活性的影响;倒置荧光显微镜和流式细胞术检测不同浓度角鲨烯对HepG2细胞内源性LDLR表达活性的影响;FRET技术检测不同浓度角鲨烯对SREBP通路中关键蛋白分子SCAP与Insig2相互作用的影响。结果角鲨烯能明显抑制HepG2细胞增殖活性,且呈浓度依赖性;倒置荧光显微镜和流式细胞术结果均表明,与对照组相比,角鲨烯能增强HepG2细胞LDLR的表达活性;FRET检测结果显示,与模型组相比,角鲨烯组YFP荧光值降低,而在5~25μmol·L^(-1)范围内,随角鲨烯浓度的增高,YFP荧光值降低。结论角鲨烯可能通过抑制SCAP/SREBP信号调控通路中关键蛋白分子SCAP与Insig2的相互作用,上调HepG2细胞LDLR的表达,是具有潜在价值的降胆固醇新药。Aim To investigate the effect of squalene on LDLR expression in HepG2 cells and its mechanism of down-regulated cholesterol.Methods The proliferation of HepG2 cells exposed to squalene at different concentrations was measured by MTT assay.The effect of squalene on the expression of LDLR in HepG2 cells was measured by flow cytometry and fluorescence microscopy.The effect of different concentrations of squalene on the interaction between SCAP and Insig2,two key protein molecules of SREBP pathway,was assayed by FRET technology.Results MTT results showed that squalene had inhibitory effect on the proliferation of HepG2 cells in a dose-dependent manner.Flow cytometry and fluorescence microscopy results showed that squalene enhanced LDLR expression in HepG2 cells compared with the control group.The results of FRET technology revealed that compared with model control group,the YFP fluorescence value in Squalene group dramatically declined,and the YFP fluorescence value of each drug group decreased with the range of 5~25 μmol·L^（-1） squalene concentration.Conclusions Squalene may promote the expression of LDLR in HepG2 cells through inhibiting the interaction between SCAP and Insig2 proteins in SREBP pathway,which may confirm that squalene is a potential novel drug for the down-regulation of cholesterol level.

关 键 词：角鲨烯 LDLR HEPG2 SREBP固醇通路 胆固醇 FRET技术 

分 类 号：R329.24[医药卫生—人体解剖和组织胚胎学] R392.11[医药卫生—基础医学]