金刚烷胺修饰物抗禽流感病毒的作用机制

目的：初步探讨金刚烷胺修饰物（NAM）抗禽流感病毒的作用机制，为抗禽流感病毒新药NAM的开发提供实验依据。方法：对狗肾细胞（MDCK）进行体外培养，分为正常对照组，病毒对照组和受试物组。①采用细胞病变法结合MTT法检测NAM对禽流感病毒（H5N1）的抑制作用，受试物组分为先加入NAM后感染病毒、先感染病毒后加入NAM和感染病毒的同时加入NAM，观察3种方式NAM对禽流感病毒的半数抑制浓度(IC50)和治疗指数(TI)；②采用神经氨酸酶抑制实验检测NAM对神经氨酸酶的活性影响。结果：① 3种不同的实验方式NAM剂量对数与细胞保护率均呈正相关关系（r 分别为0.95、0.95和0.99，P均<0.05），且NAM与保护率存在量效依赖关系。先加入NAM后感染病毒，NAM的IC50为15.32 mg·L-1，TI为103.31；先感染病毒后加入NAM，NAM的IC50为30.78 mg·L-1，TI为28.10；感染病毒的同时加入NAM，NAM的IC50为203.92 mg·L^-1，TI为4.24。②NAM具有一定的流感病毒神经氨酸酶抑制活性，其IC50为87.36 mg·L^-1。结论：NAM对穿入细胞后的禽流感病毒有较好的抑制作用，同时干扰病毒吸附和侵入细胞的脱壳过程也有一定的作用，能够在一定程度上抑制病毒的神经氨酸酶活性。

Mechanism of modified amantadine against influenza A(H5N1) virus

Objective To explore the mechanism of modified amantadine(NAM)against influenza A(H5N1) virus and to provide experimental basis for development of NAM.Methods DCK cells were cult ivated in vitro.Normal control group,virus control group and test group were included in this study.① Cytopathic effect(CPE) with MTT was used to detect the inhibitory effect of NAM against influenza A(H5N1) virus.The 50% inhibitory concentration(IC50) and therapeutic index(TI) of NAM on influenza A(H5N1) virus were studied in three ways: adding NAM before contracting virus,contracting virus before adding NAM and contracting virus with adding NAM at the same time.② The neuramidinase(NA) inhibitory effect was observed to obtain the effect of NAM on neuramidinase activity.Results ①Positive correlation was observed between logarithm of dose and protect rate of NAM in MDCK cells in the three different ways(0.95,0.95 and 0.99;all P<0.05).IC50 of NAM was 15.32 mg·L-1 and TI was 103.31 when adding NAM before contracting virus,IC50 of NAM was 30.78 mg·L-1 and TI was 28.10 when contracting virus before adding NAM,and IC50 of NAM was 203.92 mg·L-1 and TI was 4.24 when contracting virus with adding NAM at the same time.② NAM can inhibit neuramidinase activity,and IC50 was 87.36 mg·L-1.Conclusion NAM can better inhibit influenza A(H5N1) virus in cells,and has some effects on interfering viral adsorption and invading the process of cell uncoating.NAM can also inhibit neuramidinase activity to some extent.