甲型H1N1流感重组病毒模型的建立及应用

发布时间：2018-07-06 14:21

本文选题：重组病毒 + HN流感　；参考：《药物分析杂志》2016年07期

【摘要】：目的:针对流感病毒感染宿主机体的作用机制,建立以甲型H1N1流感A/California/04/2009(H1N1)为靶点的重组病毒模型,并对其应用进行探索。方法:通过将表达流感病毒血凝素蛋白HA、神经氨酸酶NA及敲除外壳基因、复制基因并带有荧光素酶报告基因的HIV~(-1)质粒共转染至293T包装细胞,产生以HA[A/California/04/2009(H1N1)]和NA[A/California/04/2009(H1N1)]为膜蛋白,HIV~(-1)(p NL4-3.Luc.R-E-)为核心的重组病毒。通过该重组流感病毒模型评价阳性对照化合物(奥斯他韦、奥斯他韦酸)、中和血清以及板蓝根药材提取液对重组病毒活性的抑制作用。结果:奥司他韦、奥司他韦酸抑制重组病毒p NL+HA04+NA04的IC50为1.07×10-8 mol·L~(-1)和1.82×10-9 mol·L~(-1);中和血清的IC50为1∶320~1∶640;中药板蓝根提取液的IC50为1.91×10-5 g·m L~(-1)。结论:奥斯他韦、奥斯他韦酸、中和血清及板蓝根提取液对于重组病毒的活性有抑制作用,提示所构建的重组病毒模型可用于上述作用机制药物的筛选和评价。
[Abstract]:Aim: to establish a recombinant virus model targeting A / H1N1 influenza A / California / 04 / 2009 (H1N1) and to explore its application. Methods: HIV ~ (-1) plasmids expressing influenza virus hemagglutinin (HA), neuraminidase (na) and knockout gene were co-transfected into 293T packaging cells by replicating genes and carrying luciferase reporter gene. Recombinant viruses were produced with HA [A / California / 04 / 2009 (H1N1)] and na [A / California / 04 / 2009 (H1N1)] as the membrane protein HIV-1) (p NL4-3.Luc.R-E-. The recombinant influenza virus model was used to evaluate the inhibitory effect of positive control compounds (oseltamivir, oseltamivir acid), neutralizing serum and Radix Isatidis extract on the recombinant virus activity. Results: the IC50 of the recombinant virus pNL HA04 NA04 was 1.07 脳 10 ~ (-8) mol / L ~ (-1) and 1.82 脳 10 ~ (-9) mol / L ~ (-1) for oseltamivir and oseltamivir acid respectively, the IC50 for neutralizing serum was 1: 320 ~ 1: 640, and the IC50 for Radix Isatidis extract was 1.91 脳 10 ~ (-5) g / L ~ (-1). Conclusion: oseltamivir, oseltamivir acid, neutralizing serum and Radix Isatidis extract can inhibit the activity of recombinant virus.
【作者单位】：中国食品药品检定研究院;
【基金】：国家药典委员会国家药品标准提高研究(670) 中国食品药品检定研究院中青年研究发展基金(2012A12) 化药制剂质量评价关键技术研究课题(2015ZX09303001) 国家自然科学基金(81303194)
【分类号】：R373.13

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