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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 BHK-21 infected by Japanese encephalitis virus (JEV). Methods BHK-21 cells were treated with various concentrations of NGF (100,50,25,12.5,6.3,3.1,1.6 and 0.8 μg / L). The toxic effects of NGF on BHK-21 cells were detected by MTT assay. The JEV (10-1, 10-2, 10-3, 10-4, 10-5, 10-6, 10-7) JEV was used to infect BHK-21 cells and the plaque count was used to determine the virus titer. Infected BHK-21 cells by adding different concentrations of NGF, CPE cells were observed, and by reducing the rate of plaque analysis of NGF on JEV inhibition. Results The titer of JEV was Lg 8.70 pfu / ml and the level of BHK-21 cells in NGF (12.5μg / L) + JEV group was significantly reduced with NGF concentration less than 100μg / L. With the increase of NGF concentration (R = 0.929, P <0.05). When the concentration of NGF was above 3.1μg / L, the reduction rate of plaque was more than 10%, and the anti-JEV effect was obvious. Conclusion NGF at a concentration range of 0.8 ~ 100μg / L has a significant inhibitory effect on JEV, which lays the foundation for future research on the mechanism of NGF antiviral activity.