以人CCR5启动子为靶的药物筛选方法的初步建立  

Establishment and Initial Application of a Medicine Screening Technique Based on Human Promoter of CCR5

作　　者：卫艳萍[1] 冯龙[1] 马云云[1] 董子明[1] 韩志强[2] 赵国强[1] 

机构地区：[1]郑州大学基础医学院,郑州450001 [2]郑州大学第一附属医院,郑州450052

出　　处：《生物医学工程学杂志》2009年第5期1043-1046,1063,共5页Journal of Biomedical Engineering

摘　　要：构建靶向人趋化因子受体5(CCchemokine receptors5,CCR5)启动子的药物筛选系统。将CCR5启动子序列克隆入改建后的报告载体pGL3-neo,转染pGL3-neo-CCR5入Jurkat细胞(急性T淋巴细胞性白血病细胞),G418筛选后分组;用7种中药分别作用于各组细胞16h后,检测细胞中CCR5启动子的表达水平。结果显示,双黄连组比对照组的荧光值明显降低(P<0.05);川琥宁组、黄芩组、黄芪组比对照组的荧光值明显升高(P<0.01)。双黄连可使体外培养细胞转染的CCR5启动子活性降低,川琥宁、黄芩、黄芪可使体外培养细胞转染的CCR5启动子活性增高。表明初步构建成功靶向人CCR5启动子的药物筛选系统。This research ws carried out to construct a medicine screening system targeting at human promoter of CCR5. The gene Human promoter of CCR5 was inserted into the rebuilt vector pGL3-neo. The pGL3-neo-CCR5 plasmids were transfected into Jurkat cells（the cell line of acute T lymphocyte leukemia）. The lasting transfected cells were screened by G418. After seven kinds of traditional Chinese medicine had acted separately on the lasting transfected cells for 16h, the expression levels of CCR5 promoter in the cells were detected. The results showed that the level of luciferase activity of Shuanghuanglian-injectio group was remarkably lower than that of control（P〈0.05）, and the levels of luciferase activity of Chuanhuning group,Baical skullcap root group, and Milkvetch root group were remarkably higher than that of control （P〈0.01）. Shuanghuanglian-injectio depressed the activity of the transfected CCR5 promoter in cells cultivated in vitro; Chuanhuning,Baical skullcap root and Milkvetch root boosted the activity of the transfected CCR5 promoter in cells cultivated in vitro. Thus a medicine screening system based on Human promoter of CCR5 was initially constructed.

关 键 词：报告载体pGL3 CCR5启动子 体外筛选 中药 

分 类 号：R965.1[医药卫生—药理学]