黄柏提取物抑制Ⅰ型单纯疱疹病毒复制机制的初步研究  

Phellodendron amurense inhibited replication of herpes simplex virus type I

作　　者：孙鹏 杨丽敏 赵鹏伟 陆景坤 张烜 红梅 陶格斯 李恋 棊鲁博 敖登托娅 王怡诺 张保平[2] Sun Peng;Yang Limin;Zhao Pengwei;Lu Jingkun;Zhang Xuan;Hong Mei;Tao Gesi;Li Lian;Qi Lubo;Odentoya;Wang Yinuo;Zhang Baoping(Basic Medical College of Inner Mongolia Medical University,Hohhot 010010,China;Department of Medical Laboratory,Affiliated Hospital of Inner Mongolia Medical University,Hohhot 010010,China;Department of Clinical Pharmacy,Inner Mongolia Medical Universityy Hohhot 010010,China)

机构地区：[1]内蒙古医科大学基础医学院,呼和浩特0100101 [2]内蒙古医科大学附属医院检验科,呼和浩特010010 [3]内蒙古医科大学临床药学系,呼和浩特010010

出　　处：《中华实验和临床病毒学杂志》2019年第1期25-29,共5页Chinese Journal of Experimental and Clinical Virology

基　　金：内蒙古自然科学基金[2016MS(LH)0818]。

摘　　要：目的研究黄柏对Ⅰ型单纯疱疹病毒(herpess implex 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HeLa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24h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、40ng/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24h后,qPCR检测显示,细胞IP-10、IL-12、IFN-γ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。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 NF-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.

关 键 词：黄柏 HSV-1 病毒囊膜表面糖蛋白 免疫因子 NF-κB(P65) 

分 类 号：R285[医药卫生—中药学]