低密度脂蛋白受体/报告基因系统建立及中药降脂药物的筛选

被引:13
作者
唐建华
高小平
张义正
机构
[1] 四川大学生命科学院
[2] 地奥制药集团药物研究所
[3] 四川大学生命科学院 成都
[4] 成都
关键词
低密度脂蛋白受体; 荧光素酶报告基因; 中药; 药物筛选;
D O I
10.16333/j.1001-6880.2005.03.017
中图分类号
TQ464 [生物制品药物的生产];
学科分类号
100705 ;
摘要
为了从中药中筛选降脂药物,本文构建了低密度脂蛋白受体/荧光素酶(LDLR/Luc)报告基因系统。将LDLR/Luc报告质粒转染至HepG2细胞,并优化细胞接种密度、细胞培养时间以及检测底物浓度等条件,获得较高荧光素酶活性;进一步选择普伐他汀作为阳性对照药物,可明显提高LDLR/Luc转录活性,表明成功地建立了靶向ldlr转录活性的报告基因筛选体系。应用该体系对500余种中药提取物进行了筛选,决明子、泽泻、穿龙薯蓣等提取物显示阳性,与过去报道采用动物模型研究的结果一致。
引用
收藏
页码:316 / 319
页数:4
相关论文
共 12 条
[1]  
Protein binding to the low density lipoprotein receptor promoter in vivo is differently affected byy gene activation in primary human cells. Jeffl,Ells worth,et al. Journal of Lipid Research . 1995
[2]  
Identification of a novel cisacting element participating in maximal induction of the human Low density lipoprotein receptor gene transcription in response to low cellular cholesterol levels. Kamal DM,Ruixin,Joey,et al. Journal of Biological Chemistry . 1996
[3]  
Development and use of a gene promoter-based screen to identify novel inhibitors of cyclooxygenase-2 transcription. Subbaramaiah K,Bulic P,Lin Y,et al. Journal of Biomolecular Screening . 2001
[4]  
Nuclear protein that binds sterol regulatory element of low density lipoprotein receptor promoter.I. Identification of the protein and delineation of its target nucleotide sequence. Briggers MR,Yokoyama C,Brown MS,et al. Journal of Biological Chemistry . 1993
[5]  
Peroxisome proliferator-activated receptor alpha is involved in the regulation of lipid metabolism by ginseng. Yoon M,Lee H,Jeong S,et al. British Journal of Pharmacology . 2003
[6]  
Inhibitors of choleserol biosynthesis increase hepatic low-density lipoprotein receptor protein degradation. Ness GC,Zhao Z,Lopez D. Archive of Biochemistry and Biophysics . 1996
[7]  
Hypolipidemic activity of seeds of Cassia tora Linn. Patil UK,Saraf S,Dixit VK. Journal of Ethnopharmacology . 2004
[8]  
LCAT modulates atherogenic piasma lipoproteins and the extent of atherosclerosis only in the presence of mormal LDL receptors in transgenic rabbits. Broussean ME,Kauffman RD,Herderick EE. Arteriosclerosis and Thrombosis . 2000
[9]  
Effects of Diosgenin on cholesterol Metabolism. Laguna J,Gomcz-Puyou A,Pena A,et al. Journal of Atherosclerosis Research . 1962
[10]  
A simple statistical parameter for use in evaluation and validation of high throughput screening assays. Zhang JH,Chung TD,Oldenburg KR. Journal of Biomolecular Screening . 1999