论文部分内容阅读
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。
OBJECTIVE: To establish a recombinant virus model targeting A / H1N1 influenza A / California / 04/2009 (H1N1) targeting to the mechanism of influenza virus infection in host organism and to explore its application. Methods: The 293T packaging cells were co-transfected with HIV ~ (-1) plasmid expressing the influenza virus hemagglutinin HA, neuraminidase NA and the knockout shell gene, and the luciferase reporter gene. (-1) (p NL4-3.Luc.RE-) with HA [A / California / 04/2009 (H1N1)] and NA [A / California / 04 / As the core of the recombinant virus. The recombinant influenza virus model was used to evaluate the inhibitory effect of the positive control compound (oseltamivir and oseltamivir), the neutralizing serum, and the extract of Radix isatidis on recombinant virus activity. Results: The results showed that oseltamivir and oseltamivir inhibited IC50 of recombinant virus p NL + HA04 + NA04 1.07 × 10-8 mol·L -1 and 1.82 × 10-9 mol·L -1 ). The IC50 of neutralizing serum was 1: 320 ~ 1: 640. The IC50 of Radix isatidis was 1.91 × 10-5 g · m L -1. CONCLUSION: Ostevastatin, oseltamivir, neutralizing serum and extract of Banlangen can inhibit the activity of recombinant virus, suggesting that the constructed recombinant virus model can be used for the screening and evaluation of the above mentioned mechanism of action.