爪蟾卵母细胞固醇调控报告系统的建立及中药调脂作用研究  

Establishment of Sterol Regulatory Report System in Xenopus Oocytes and its Application for Screening Hopolipidemic

作　　者：范春雷[1] 钱颖[1] 卢德赵[1] 沃兴德[1] 高丽萍[1] 严瑾 

机构地区：[1]浙江中医药大学生命科学院,杭州310053 [2]杭州师范学院基础医学部,杭州310036

出　　处：《中国药学杂志》2006年第9期664-667,共4页Chinese Pharmaceutical Journal

基　　金：国家自然科学基金(30070932);浙江省自然科学基金(Y204269);浙江省教育厅科研基金(0424XP25)

摘　　要：目的在爪蟾卵母细胞中建立低密度脂蛋白受体基因表达的固醇调控报告系统,通过中药的干预培养,从基因调控水平上阐明中药调脂作用的机制,并为药物筛选提供一个可靠的实验平台。方法将绿色荧光蛋白报告基因构建在调控低密度脂蛋白受体基因表达的固醇调控元件的下游,经显微注射导入爪蟾卵母细胞,在分别用含姜黄素、茶多酚等中药的培养液中培养3d后,用荧光显微镜及荧光分析仪对绿色荧光蛋白的表达情况进行定性定量分析。结果姜黄素诱导了绿色荧光蛋白的表达。结论①姜黄素可以通过影响固醇调控元件的作用上调低密度脂蛋白受体基因的表达；②建立的固醇调控报告系统可作为降胆固醇药物筛选和作用机制研究的实验平台。OBJECTIVE To establish a sterol regulatory report system of LDLR gene expression in Xenopus oocytes, investigate the lipidlowering mechanism of Chinese medicine and provide a stable experimental system for Hopolipidemic. METHODS A green fluorescence protein （GFP） gene in the downstream of sterol regulatory element-1 was constructed, a plasmid named pLXRN-4SRE-fPA was abtained. This plasmid was introduced into the nucleus of Xenopus oocytes by microinjection; the oocytes were cultured in the medium containing different kinds of Chinese medicines such as curcumin, tea polyphenols, tanshinone, and ligustrazine, etc. After 3 d, the expression of GFP in the oocytes was detected by a fluorescence microscope and GFP fluorescence analyzer. RESULTS The expression of GFP was obviously induced only in the cureumin group. CONCLUSION ①Curcumin can obviously up-regulate the expression of LDLR gene by influencing on SRE-1 ; ②The sterol regulatory report system establishes in the Xenopus oocytes can be a system for study on the lipid-lowering mechanism of Chinese medicine and screening Hopolipidemic.

关 键 词：爪蟾卵母细胞 固醇调控元件 低密度脂蛋白受体 绿色荧光蛋白 姜黄素 

分 类 号：R203.4[医药卫生—中医学] R285.3