摘要
目的建立并优化VSVG/HIV-1_(NL4-3)Luc假病毒筛选抗HIV药物药效模型。方法参照Promega公司的荧光素酶活性分析系统,比较VSVG/HIV-1_(NL4-3)Luc对4种不同细胞的感染力。通过采用3种不同实验条件对不同类型HIV-1上市药物进行活性评价,进一步验证了该模型的可行性与有效性。最后,应用该系统对特定靶点化合物进行筛选,并与HIV-1_(ⅢB)病毒筛选体系所得结果进行比较。结果 VSVG/HIV-1_(NL4-3)Luc对CRFK细胞的感染力最强,报告基因的表达水平与加入的病毒量呈剂量依赖关系。比较3种实验条件下不同阳性药物抑制VSVG/HIV-1_(NL4-3)Luc的半数有效剂量浓度EC50,发现第3种条件最优。而用该假病毒对合成化合物进行筛选,发现其EC50与病毒HIV-1_(ⅢB)所得EC_(50)基本一致。结论成功建立并优化了基于VSVG/HIV-1_(NL4-3)Luc假病毒的抗HIV药物筛选系统。
Aim To establish and optimize the VSVG/HIV-1_(NL4-3)Luc pseudovirus model for anti-HIV drugs screening.Methods The infectivity of VSVG/HIV-1_(NL4-3)Luc in 4 different cell lines was investigated according to the method of the luciferase activity analysis system of Promega company. 3 different experimental settings were used to detect the activities of approved anti-HIV drugs to confirm the feasibility and effectiveness of the system. Finally,some potential compounds were screened for their anti-HIV activities,and their antiviral activities against the pseudovirus were compared with HIV-1_(ⅢB). Results The pseudovirus showed the strongest replication ability in CRFK cells,and a clear dose-effect relationship was found between the report gene expression level and the virus quantity. Comparing the EC_(50) of different positive inhibitors against VSVG/HIV-1_(NL4-3)Luc on 3 kinds of experimental conditions,3rd scheme is the best.Finally,the system was used to screen compounds,the EC_(50)s against pseudovirus were similar to those in HIV-1_(ⅢB). Conclusion An optimized VSVG/HIV-1_(NL4-3)Luc anti-HIV screening system has been successfully developed.
出处
《中国药理学通报》
CAS
CSCD
北大核心
2016年第3期433-438,共6页
Chinese Pharmacological Bulletin
基金
国家科技重大专项课题(No 2009ZX09501-029
2014ZX10005-002)
国家自然科学基金资助项目(No 81471620
81102483)
云南省应用基础研究计划(No 2015FB182)
