建立一个基于报告基因的靶标筛药系统筛选上调ABCA1基因转录的中药提取物  

Establishment of a Reporter Gene-Based Drug Screening System for Identifying Herbal up-Regulators of ATP-Binding Cassette Transporter A1

作　　者：胡蔚蓉[1] 连霁虹[1] 纪伟[1] 郭雷[1] 龚邦强[1] 

机构地区：[1]华东理工大学生物反应器工程国家重点实验室,上海200237

出　　处：《华东理工大学学报（自然科学版）》2007年第6期784-787,882,共5页Journal of East China University of Science and Technology

摘　　要：ATP结合盒转运蛋白A1(ABCA1)具有催化外周细胞过量胆固醇和磷脂外向流出的功能。本实验将ABCA1启动子序列克隆到含有荧光素酶报告基因和新霉素抗性基因的pGL3B-neo载体中,并将得到的重组质粒pGL3B-neo/ABCA1转染入人肝癌细胞(hepG2)。通过稀释培养法最终得到含有重组质粒的稳定细胞系SABCA1。利用这个报告基因为基础的药物筛选系统,筛选了100多种中药提取物。通过荧光素酶活性测试,相对活性达到180%以上的有4种中药提取物,并最终挑选能明显激活ABCA1基因启动子的鬼箭羽水提组分进一步研究,得出半数有效浓度(EC50)为48.06 mg/L。此外,利用基于apoA1基因的药物筛选系统验证得出鬼箭羽水提组分也能促进apoA1基因的表达。ABCAl promotes the cholesterol and phospholipid efflux from peripherial cells. The promoter of ATP-binding cassette transporter Al gene was cloned into pGL3B-neo vector containing luciferase gene and neomycin resistance gene. The recombinant reporter gene vector pGL3B-neo/ABCAl was then transfected into hepG2 cells and therefore the stable cell line SABCAl was obtained. Using this cell-based drug screening system, more than 100 herbal extracts were screened. Through luciferase activity testing, four extracts, shown more than 180G relative activity on SABCAl cells, were regarded as active samples and the water extract of Euonymus alatus （Thunb.） Sieb. was identified and its EC50 value was tested. Furthermore, an apoAl gene based drug screening system was also used to confirm that the water extract of Euonymus alatus （Thunb.） Sieb. is capable of activating apoAl gene expression. This herbal medicine extract is eikely to be an effective novel medicine for atheromatosis.

关 键 词：TP结合盒转运子Al 动脉粥样硬化 高密度脂蛋白 胆固醇逆向转运 

分 类 号：TQ785[化学工程]