神经生长因子对乙型脑炎病毒感染BHK-21细胞的抑制作用  

Inhibitory effect of nerve growth factor on infection of BHK-21 cells with Japanese encephalitis virus

作　　者：杨栋[1] 顾鹏毅[1] 刘海军[1] 李宁禾[1] 刘畅[2] 姚宇[2] 李鹤[3] 孙英杰 孙晋民[1] 

机构地区：[1]中国医科大学实验技术中心一部暨国家中医药管理局中药生物工程科研Ⅲ级实验室,辽宁沈阳110001 [2]沈阳药科大学生命科学与生物制药学院,辽宁沈阳110016 [3]辽宁大学生命科学学院,辽宁沈阳110036 [4]辽宁成大生物股份有限公司,辽宁沈阳110176

出　　处：《中国生物制品学杂志》2013年第6期840-843,共4页Chinese Journal of Biologicals

基　　金：辽宁省科技基金项目(2008225021-1)

摘　　要：目的观察神经生长因子(nerve growth factor,NGF)对乙型脑炎病毒(Japanese encephalitis virus,JEV)感染地鼠肾细胞BHK-21的抑制作用。方法将不同浓度的NGF(100、50、25、12.5、6.3、3.1、1.6和0.8μg/L)加入BHK-21细胞中,采用MTT法检测NGF对BHK-21细胞的毒性作用;将不同稀释度的JEV(10-1、10-2、10-3、10-4、10-5、10-6、10-7)JEV感染BHK-21细胞,蚀斑计数法测定病毒滴度;在JEV感染的BHK-21细胞中加入不同浓度的NGF,观察细胞的CPE情况,并通过蚀斑减少率分析NGF对JEV的抑制作用。结果 NGF浓度在100μg/L以内对BHK-21细胞无毒性;JEV的滴度为Lg 8.70 pfu/ml;NGF(12.5μg/L)+JEV组BHK-21细胞病变程度明显减轻;随着NGF浓度的提高,蚀斑减少率也呈升高趋势(r=0.929,P<0.05),NGF浓度为3.1μg/L以上时,蚀斑减少率达10%以上,具有明显的抗JEV效果。结论 NGF在浓度为0.8～100μg/L的范围内,对JEV具有明显的抑制作用,为今后研究NGF抗病毒的作用机制奠定了实验基础。Objective To observe the inhibitory effect of nerve growth factor（NGF）on infection of BHK-21 cells infected with Japanese encephalitis virus（JEV）.Methods BHK-21 cells were treated with NGF at various concentrations（100,50,25,12.5,6.3,3.1,1.6 and 0.8 μg/L）respectively.The toxic effect of NGF on BHK-21 cells was determined by MTT method.BHK-21 cells were infected with JEV at various dilutions（10-1,10-2,10-3,10-4,10-5,10-6 and 10-7）,and determined for titer by plaque counting.BHK-21 cells infected with JEV were treated with NGF at various concentrations and observed for CPE,based on which the inhibitory effect of NGF on JEV was analyzed by plaque reduction rate.Results NGF at concentrations of less than 100 μg/L showed no cytotoxic effect on BHK-21 cells.The titer of JEV was Lg8.70 pfu/ml.The CPE of BHK-21 cells in NGF（12.5 μg/L）＋ JEV group was relieved significantly.The plaque reduction rate showed an increasing tendency with the increased NGF concentration（r = 0.929,P ＆lt; 0.05）.The plaque reduction rate of BHK-21 cells treated with NGF at concentrations of more than 3.1 mg/L was above 10%,indicating significantly inhibitory effect on JEV.Conclusion The NGF at concentrations of 0.8 - 100 μg/L showed significantly inhibitory effect on JEV,which laid an experimental foundation of further study on mechanism of antiviral effect of NGF.

关 键 词：神经生长因子 乙型脑炎病毒 抑制作用 BHK-21细胞 

分 类 号：Q575.11[生物学—生物化学] R373.31[医药卫生—病原生物学]