大黄酚对Huh-7细胞SREBP表达及脂质代谢的影响  

Effects of chrysophanol on expression of SREBPs and lipid metabolism in Huh-7 cells

作　　者：李金梅[1,2] 丁丽丽[2] 宋保亮[3] 杨莉[2] 王峥涛[1,2] 

机构地区：[1]中国药科大学,江苏南京211198 [2]上海中医药大学中药研究所中药标准化教育部重点实验室暨上海市复方中药重点实验室,上海201203 [3]中国科学院上海生命科学研究院生物化学与细胞生物学研究所分子生物学国家重点实验室,上海200031

出　　处：《药学学报》2015年第2期174-179,共6页Acta Pharmaceutica Sinica

基　　金：国家自然基金优秀青年基金项目(81222053);国家自然基金青年基金项目(81303186);中国博士后基金项目(2013M531202);一流学科(A类)中药学学科创新基金(ZYX-NSFC-016)

摘　　要：大黄具有泻下、降脂和减肥等作用,但其降脂药效物质及作用机制尚不清楚。本文探讨大黄的主要活性成分大黄酚(chrysophanol)对人肝癌细胞株(human liver carcinoma Huh-7 cells,Huh-7)固醇调节元件结合蛋白(sterol regulatory element-binding proteins,SREBPs)表达及脂质代谢的影响。以双荧光素酶报告基因检测试剂盒检测大黄酚对SREBP的转录抑制作用;总胆固醇(total cholesterol,TC)和甘油三酯(triglyceride,TG)测定试剂盒检测细胞内TC、TG水平;荧光定量RT-PCR法检测SREBP及靶基因m RNA的表达;采用CCK-8试剂盒检测细胞存活率。结果显示,经大黄酚(40μmol·L-1)干预16 h后,使细胞内TC、TG水平显著减少;大黄酚可以抑制SREBP转录,并呈现出一定的浓度依赖性;能显著下调SREBP及靶基因的表达;大黄酚对细胞活力无明显影响。结果提示,大黄酚可能通过下调肝细胞SREBP及靶基因的表达,从而降低细胞内TC、TG的含量,减轻肝细胞脂肪变性,起到改善脂质代谢的作用。Rhubarb is a traditional Chinese medicines which possess laxative, lipid-lowering, and weight- loss activities, but the active compounds of lipid-lowering and underlying molecular mechanisms are not yet clear. This study aims to explore the effects of chrysophanol on the m RNA expressions of sterol regulatory element-binding proteins（SREBPs） and lipid metabolism in human liver carcinoma Huh-7 cells, which is one of the active compounds obtained from Rhubarb. A reporter gene assay was used to test the transcription of SREBP. The intracellular triglyceride and total cholesterol contents were measured by using commercially available test kits. The SREBPs target genes expressions were measured by Quantitative Real-Time PCR. Cell viability was evaluated by Cell Counting Kit-8. As the results shown, chrysophanol（40 μmol·L-1, 16 h） could notably inhibited human SRE promoter activity in a dose-dependent manner and decrease intracellular cholesterol and triglyceride levels. Furthermore, the m RNA expressions of SREBPs target genes were significantly downregulated by chrysophanol treatment. However there are no significant differences on cell viability when compared with the control group. These results suggested that chrysophanol might improve lipid metabolism through suppressing the m RNA expressions of SREBPs target genes to attenuate intracellular lipid accumulation.

关 键 词：大黄酚 大黄 固醇调节元件结合蛋白 总胆固醇 甘油三酯 HUH-7细胞 

分 类 号：R963[医药卫生—微生物与生化药学]