黄柏提取物抑制Ⅰ型单纯疱疹病毒复制机制的初步研究

摘要

Objective To study the effects of Phellodendron amurense on herpes simplex virus 1 (HSV-1) and cytokines,and to explore the mechanism of Phellodendron amurense inhibiting HSV-1 virus through multiple channels.Methods Viruses were inoculated into medicine treated HeLa cells.The proliferation of virus was observed by fluorescence microscopy.The transcriptional levels of glycoprotein gD and functional protein US1 on the surface of virus envelope were detected by quantitative (q) PCR.After incubating HeLa cells for 24 h,qPCR was used to detect the expression of intrinsic immune factors such as IP-10,IL-12,IFN-gamma and transcription factor N F-kappa B (P65).The expression and nuclear location of NF-kappa B (P65) protein were detected by immunofluorescence.Results Fluorescence showed that the proliferation of virus decreased significantly at 8 and 40 ng/ml (P＜0.01),and the transcription levels of viral protein gD and US1 decreased (P＜0.05).After incubation for 24 hours,the transcription levels of IP-10,IL-12 and IFN-gamma in HeLa cells increased significantly (P＜0.01).The transcription level of transcription factor NF-kappa B (P65) also increased (P＜0.05),and immunofluorescence showed that the nuclear penetration rate of p65 subunit of NF-kappa B (P65) in each group increased (P ＜ 0.05).Conclusions Phellodendron amurense extract can inhibit HSV-1 by inhibiting the transcription of viral functional protein and promoting the expression of cellular immune factors.%目的 研究黄柏对Ⅰ型单纯疱疹病毒(herpes simplex virus 1,HSV-1)功能蛋白和其对细胞因子的作用,探讨黄柏多渠道抑制HSV-1病毒的机制.方法 将病毒接种到药物处理后的HLLa 细胞上,荧光显微镜观察病毒增殖变化,qPCR检测病毒囊膜表面糖蛋白gD与功能蛋白US1的转录水平表达.药物孵育HeLa细胞24 h后,qPCR检测IP-10、IL-12、IFN-γ等固有免疫因子转录水平表达变化.免疫荧光检测NF-κB(P65)核定位的情况.结果 荧光显示病毒在8、40 n/ml药物浓度下增殖显著减少(P＜0.01);且病毒蛋白gD与US1转录水平下降(P＜0.05).药物孵育HeLa细胞24 h后,qPCR检测显示,细胞IP-10、IL-12、IFN-y等转录水平显著升高(P＜0.01).同时免疫荧光显示各组NF-κB(P65)入核率有所上升(P＜0.05).结论 黄柏提取物可通过抑制病毒功能蛋白转录及促进细胞免疫因子表达来抑制HSV-1.