靶向HIV-1启动子的药物筛选系统的建立

摘要

Aim:To construct a medicine screening system targeting on HIV-1 promoter. Methods:The gene of HIV-1 promoter was inserted into the luciferase reporter vector pGL4. 17,and the plasmid of pGL4. 17-HIV-1P was transfected into H9 cells. The transfected cells were screened by G418. Four kinds of traditional Chinese medicine (Eucommia Bark, Baical Skullcap Root,Lightyellow Sophora Root,and Shuanghuanglian) decoction were given to rats intragastrically and the medicated serum were collected. After the stable transfected H9 cells were stimulated by the four kinds of medicated serum, luciferase activity of each groups were detected. Results: There were significant difference in luciferase activity among these groups ( F = 1 820. 333 , P < 0. 001 ). The Shuanghuanglian could depress the luciferase activity remarkably ( P < 0. 05 ), Eucommia Bark, Baical Skullcap Root and Lightyellow Sophora Root could enhance it (P <0. 05). Conclusion; A medicine screening system based on HIV-1 promoter is established successfully and Shuanghuanglian may have the potential an-ti-HIV-1 effect.%目的:建立靶向HIV-1启动子的药物筛选系统.方法:将HIV-1启动子核心序列克隆入荧光素酶报告基因载体pGLA.17中,构建重组质粒pGL4.17-HIV-1P并转染H9细胞;分别用杜仲、黄芩、苦参及双黄连灌胃大鼠后采血,将含药血清分别作用于转染的H9细胞,检测H9细胞荧光素酶活性.以灌胃生理盐水的大鼠血清处理的转染细胞为对照,以转染空质粒pGLA.17的H9细胞为空白对照.结果:6组荧光素酶活性差异有统计学意义(F=1 820.333,P＜0.001).双黄连组转染细胞荧光素酶活性降低(P＜0.05),杜仲、黄芩及苦参组转染细胞荧光素酶活性增高(P＜0.05).结论:初步构建了靶向HIV-1启动子的药物筛选系统,并推测双黄连具有抗HIV-1作用.