去泛素化酶抑制剂PR-619抑制单纯疱疹病毒复制机制的研究

The mechanism study of a deubiquitinating enzyme inhibitor PR-619 on suppression herpes simplex virus replication

目的 :探讨去泛素化酶抑制剂PR-619对人类单纯疱疹病毒(herpes simplex virus,HSV)复制的影响。方法 :通过In-cell Western和Western blot技术检测病毒复制及蛋白表达,同时利用real-time PCR检测相关病毒基因的表达水平。着重考察广谱去泛素化酶抑制剂PR-619对病毒m RNA转录和病毒相关蛋白合成的作用。结果:在细胞感染体系中,PR-619可以抑制HSV极早期及晚期蛋白(ICP0、ICP4和g D)的表达,从而有效阻断病毒的复制;进一步研究发现,PR-619可以增加细胞内泛素连接蛋白含量,从而抑制细胞内泛素循环,这可能是PR-619抑制病毒复制的机制。结论:HSV的复制依赖于功能化泛素的存在,而抑制细胞去泛素化酶(DUBs)活性可以阻断细胞内正常的泛素循环,从而抑制HSV复制;该研究为寻找抗HSV治疗药物或生殖道杀微生物剂新靶点提供了新方向。

Objective：To investigate the effect of a small molecule deubiquitinating enzyme inhibitor,PR-619 on Herpes simplex virus（HSV） replication. Methods：HSV replication and viral protein expression were detected via In-cell Western and Western blot.Viral gene expression on m RNA level was evaluated using Real-time PCR. We studied the inhibitory effect of PR-619 on viral replication,including viral m RNAs transcription and proteins expression. Results：PR-619 down-regulated HSV immediate-early and late gene expression（ICPO,ICP4 and g D） to inhibit viral replication effectively; PR-619 increased amount of ubiquitin-conjugates to suppress intracellular ubiquitin cycle,which may cause inhibitory effect on viral replication.and block viral induced NF-κB pathway activation. Conclusion：HSV effective replication is dependent on functional ubiquitin. Deubiquitinating enzymes is vital for intracellular ubiquitin cycle,which is associated to viral replication. This study could give deep insight into finding novel anti-HSV agents or genital microbicides.