一氧化氮供体和他汀对B组柯萨奇病毒感染的实验研究  

Effects of nitric oxide donors and statins on coxsackievirus B3 replication in vitro

作　　者：史巧云[1] 钟照华[1] 李晓波[1] 孙艳影[1] 张中海[1] 张凤民[1] 谷鸿喜[1] 

机构地区：[1]哈尔滨医科大学基础医学院微生物教研室,150081

出　　处：《中华微生物学和免疫学杂志》2007年第7期598-602,共5页Chinese Journal of Microbiology and Immunology

基　　金：黑龙江省教育厅科学技术研究项目(11511159);哈尔滨市科技攻关计划项目(2006AA9CS116-16);中国博士后科学基金(LBR04-230)

摘　　要：目的研究硝酸酯类和他汀类等一氧化氮(NO)供体的抗柯萨奇病毒B组3型(CVB3)的作用及其特点和机制。方法用TCID_(50)和空斑形成实验测定CVB3的毒力;MTT法确定NO供体药物的无毒性浓度;利用细胞病变效应(CPE)抑制实验和空斑形成抑制实验分析NO供体药物对CVB3在HeLa细胞和ECV-304细胞中增殖的抑制作用;并分析硝酸甘油(GTN)不同给药次数、NO浓度变化以及NO浓度与GTN对CVB3抑制效应间的相关性。结果硝酸酯类药物GTN、硝酸异山梨酯可明显抑制CVB3所致的CPE及空斑形成(P<0.05),他汀类药辛伐他汀、洛伐他汀均未显示抑制CVB3所致的CPE(P>0.05);CVB3接种前预先与GTN作用、CVB3接种同时加入GTN两种条件下CPE抑制率差异无统计学意义(P>0.05);CVB3攻击后多次给予GTN的组间CPE抑制率差异无统计学意义(P>0.05),但在CVB3攻击前不同时间点给予GTN的组间CPE抑制率差异有统计学意义(P<0.05);NO浓度与不同时间点给予GTN的CPE抑制结果呈正相关(r=0.97,P<0.01)。结论NO供体类药物硝酸酯类具有明确的抗CVB3感染作用,其抗CVB3增殖的作用与NO浓度呈正相关;他汀类药物在本实验条件下未观察到抗CVB3增殖作用,原因可能是细胞类型与他汀不匹配。Objective To investigate the effect of nitric oxide （NO） donor nitrates and statins on coxsackievirus group B type 3 （CVB3） and the related mechanism. Methods CVB3 were quantitated by 50% tissue culture infective dose （TCID50） and plaque forming test （PFU）. Two nitrates, glyceryl trinitrate （GTN） and isosorbide dinitrate （ISDN）, and two statins, lovastatin and simvastatin were studied. The cytotoxicities of the nitrates and the stafins were examined by MTT assay. The cytopathie effect （CPE） inhibitory assay and plaque reduction test were used to evaluate the antiviral effects of the nitrates and the statins on CVB3. The CPE inhibitory rate in HeLa cells treated with CVB3 and GTN simultaneously and of HeLa cells pretreated with GTN and attacked by CVB3 later was compared. The concentrations of NO produced by GTN in HeLa cells were also determined, the correlation between NO concentrations and the CPE inhibitory rates caused by different GTN administrations were analyzed. Results Both CPE and plaque forming of CVB3 were significantly inhibited by nitrates （ P 〈 0.05）, but not by statins （P 〉 0.05）. There was no significant difference between the CPE inhibitory rate in HeLa cells pretreated with GTN before CVB3 attacking and in HeLa cells treated with GTN and CVB3 simultaneously （ P 〉 0.05）. The same result was obtained in the comparison of CPE inhibitory rates in cells given GTN repeatedly （P 〉 0.05）. There were significant differences among the HeLa cells pretreated with GTN at different time points （ P 〈 0.05）; NO concentrations and the CPE inhibitory rate was positively correlated （r = 0.97, P 〈 0.01 ）. Conclusion The nitrates exhibited antiviral activity on CVB3 in HeLa cells, and the inhibitory effect of the nitrates against CVB3 was significantly related to the output of NO. Statins did not show any anti-CVB3 in HeLa or ECV- 304 cells in this study, probably due to the incompatibility between the studied cells and statins.

关 键 词：一氧化氮供体 柯萨奇病毒B组3型 硝酸酯类 他汀 

分 类 号：R373[医药卫生—病原生物学]