人二型大麻受体激动剂高通量筛选模型的建立  

Establishment of a high throughput screening model of cannabinoid receptor 2 agonist

作　　者：刘媛[1] 董志[1] 肖晓秋[2] 唐红[1] 王乐乐[1] 程玉洁[1] 

机构地区：[1]重庆医科大学药学院药理学教研室,重庆市生物化学与分子药理学重点实验室 [2]重庆医科大学生命科学研究院,重庆400016

出　　处：《细胞与分子免疫学杂志》2012年第2期194-196,共3页Chinese Journal of Cellular and Molecular Immunology

基　　金：国家自然科学基金面上项目(81173066);国家科技重大专项重大新药创制(2010ZX09401-306-1-1);重庆市自然科学基金重点项目(CSTC2009BA5086)

摘　　要：目的:基于二型大麻受体(CB2)的信号传递通路,建立体外高通量筛选CB2受体激动剂的药物模型。方法:将目的基因质粒pIRES2-EGFP-CB2、报告基因质粒pGL4.29[luc2P/CRE/Hygro]、内参质粒PRL-TK共转染CHO细胞,通过检测双荧光素酶报告基因的表达水平变化来筛选人CB2受体的激动剂。并对加入激动剂的浓度和作用时间进行优化及考察模型的稳定性。结果:给予10μmol/L JWH-015 6 h后能取得最大相对诱导率。成功建立CB2受体激动剂的高通量筛选模型,筛选模型Z'因子为0.81,表现为良好的稳定性。结论:成功地建立了CB2受体激动剂的筛选模型,为从中药中高通量筛选有效物质奠定了良好的基础。AIM： To establish a drug screening model of CB2 agonist in vitro based on signal regulation pathway.METHODS： Plasmid pIRES2-EGFP-CB2,pGL4.29[luc2P/CRE/Hygro] and PRL-TK were co-transfected into CHO cells in 96 wells plate,to screen agonists of CB2 receptor by detecting the expressing levels of dual luciferase activity.The concentration and acting time of the agonist were optimized and the stability of the model were investigated.RESULTS： The largest relative induction activity was obtained after 8 h drug administration.Establishment of a high throughput screening model for CB2 receptor agonist.The Z′ factor is 0.75 demonstrating its perfect stability.CONCLUSION： Sucessfully establish a drug screening model of CB2 agonist,which provided a basis for searching valid material from traditional Chinese medicine.

关 键 词：CB2受体 报告基因 高通量筛选模型 

分 类 号：R965.1[医药卫生—药理学]