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目的探讨琼枝麒麟菜多糖(EGP)抗单纯疱疹病毒2型(HSV-2)的活性及其作用机制。方法联合采用观察细胞病变效应(CPE)法和MTT法评价EGP对Vero细胞的毒性。空斑减数实验检测EGP对HSV-2的综合抗病毒活性、直接灭活作用、吸附及进入细胞过程的抑制作用、和感染后的治疗作用。Real-time PCR(RT-PCR)检测EGP对HSV-2基因表达及其DNA复制的影响。结果 EGP浓度低于125μg/mL对Vero细胞无毒性。EGP具有较强的综合抗病毒活性,在实验所用最低浓度(1.95μg/mL)下,EGP可达到80%的HSV-2病毒抑制率。EGP对HSV-2具有很强的直接灭活和抑制吸附作用,治疗作用相对较弱。同时,EGP可以下调HSV-2早期基因UL52和晚期基因UL27的表达,但是对病毒的DNA复制过程无影响。结论麒麟菜多糖具有显著的抗HSV-2感染活性,主要通过干扰病毒对细胞的粘附、侵入而发挥作用。
Objective To investigate the activity and mechanism of EGP on herpes simplex virus 2 (HSV-2). Methods The cytotoxicity of EGP on Vero cells was evaluated by observing cytopathic effect (CPE) and MTT assay. Plaque reduction experiments were performed to detect the combined antiviral activity of EGP against HSV-2, direct inactivation, inhibition of adsorption and entry into the cell process, and post-infection therapeutic effects. Real-time PCR (RT-PCR) was used to detect the effect of EGP on HSV-2 gene expression and DNA replication. Results EGP concentrations below 125 μg / mL were non-toxic to Vero cells. EGP has a strong comprehensive antiviral activity, at the lowest concentration used in the experiment (1.95 μg / mL), EGP achieves 80% inhibition of HSV-2 virus. EGP on HSV-2 has a strong direct inactivation and inhibition of adsorption, the treatment effect is relatively weak. At the same time, EGP can down-regulate the expression of HSV-2 early gene UL52 and late gene UL27, but it has no effect on DNA replication. Conclusion Eucheuma polysaccharides have significant anti-HSV-2 infection activity, mainly through interfering with the virus on cell adhesion, invasion and play a role.