金刚烷胺修饰物抗禽流感病毒的作用机制

J4 ›› 2009, Vol. 35 ›› Issue (5): 866-869.

金刚烷胺修饰物抗禽流感病毒的作用机制

徐坤¹, 杨松涛², 张士尧¹, 侯雷¹, 王宏芳¹, 浦昀¹, 李金华¹, 夏咸柱², 李娟¹, 李静¹

1.吉林大学公共卫生学院卫生检验教研室|吉林长春 130021；2. 解放军军事医学科学院军事兽医研究所|吉林长春 130062

收稿日期:2009-06-01 出版日期:2009-09-28 发布日期:2009-09-28
通讯作者: 李娟;李静 E-mail:li_juan@jlu.edu.cn;li_jing@jlu.edu.cn
作者简介:徐坤（1979-）|男|黑龙江省海伦市人|在读医学博士|主要从事化合物抗病毒研究。
基金资助:
吉林省科技厅科研基金资助课题（200705389）；吉林省科技厅农业重大科技专项资助课题（20040202-3-1）；吉林省高科技产业发展项目资助课题；全军医学科研“十五”计划第二批资助课题（04Z016）

Mechanism of modified amantadine against influenza A(H5N1) virus

XU Kun¹, YANG Song-Tao¹, ZHANG Shi-Yao¹, HOU Lei¹, WANG Hong-Fang¹, PU Yun¹, LI Jin-Hua¹, XIA Xian-Zhu², LI Juan¹, LI Jing¹

1.Department of Health Laboratory, School of Public Health, Jilin University, Changchun 130021, China; |2. Institute of Military Veterinary, Academy of Military Medical Sciences, Changchun 130062, China

Received:2009-06-01 Online:2009-09-28 Published:2009-09-28

摘要/Abstract

摘要：

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。

关键词: 禽流感；金刚烷胺；流感病毒A型, H5N1亚型

Abstract:

Abstract: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.

Key words: influenza virus, amantadine, influenza A virus,H5N1 subtype

中图分类号:

R978.7

徐坤, 杨松涛, 张士尧, 侯雷, 王宏芳, 浦昀, 李金华, 夏咸柱, 李娟, 李静. 金刚烷胺修饰物抗禽流感病毒的作用机制[J]. J4, 2009, 35(5): 866-869.

XU Kun, YANG Song-Tao, ZHANG Shi-Yao, HOU Lei, WANG Hong-Fang, PU Yun, LI Jin-Hua, XIA Xian-Zhu, LI Juan, LI Jing.
Mechanism of modified amantadine against influenza A(H5N1) virus[J]. J4, 2009, 35(5): 866-869.