去泛素化酶抑制剂b-AP15抑制马立克氏病病毒的复制  

Deubiquitinase inhibitor b-AP15 suppresses the replication of Marek’s disease virus in chicken cells

作　　者：于翀 王文婧[1] 谢知航 周正轩 王婷[1] 范青松 王梦涵 吕岩[1] 许家翠 艾永兴[1] YU Chong;WANG Wenjing;XIE Zhihang;ZHOU Zhengxuan;WANG Ting;FAN Qingsong;WANG Menghan;LYU Yan;XU Jiacui;AI Yongxing(Key Laboratory of Zoonosis Research,Ministry of Education,College of Animal Science,Jilin University,Changchun 130062,China)

机构地区：[1]吉林大学动物科学学院人兽共患病研究教育部重点实验室,吉林长春130062

出　　处：《中国兽医学报》2021年第10期1894-1901,共8页Chinese Journal of Veterinary Science

基　　金：吉林省自然科学基金资助项目(20180101244JC);国家自然科学基金资助项目(31272528);吉林大学国家级大学生创新创业训练计划资助项目(202110183333)。

摘　　要：为了探寻对马立克氏病病毒(MDV)增殖具有靶向抑制作用的药物,本研究以MDV编码的去泛素化酶UL36为靶点,利用去泛素化反应体系,对2448个化合物进行筛选,应用泛素荧光底物分析抑制物对UL36的抑制动力学,并应用MDV致鸡胚成纤维细胞(CEF)病变模型(CPE)和MDV致鸡马立克氏病(MD)模型分析抑制物对MDV在鸡细胞中复制的影响。结果发现,化合物b-AP15在高于5μmol/L浓度时可显著地非竞争性抑制3 nmol/L UL36的酶活性(P<0.01)。b-AP15对CEF的安全浓度小于1μmol/L,0.25,0.50μmol/L的b-AP15可以显著抑制MDV在CEF上的CPE形成以及在CEF细胞内的复制(P<0.01)。以2,5 mg/(kg·d)b-AP15对MDV攻毒雏鸡通过腹腔给药治疗,可显著抑制MDV在鸡T细胞内的复制(P<0.01)。这些结果说明,去泛素化酶抑制剂b-AP15可抑制MDV在鸡细胞内的复制。To obtain the drugs inhibiting the proliferation of Marek?s disease virus(MDV),the MDV-encoded deubiquitinase UL36 was used as the target for screening 2448 compounds with a deubiquitination reaction system.The UL36 inhibition kinetics was conducted utilizing different fluorescent substrate concentration in the absence or presence of the screened out compound.Cytopathic effect(CPE)induced by MDV on chicken embryo fibroblast(CEF)and the chickens challenged by MDV were used to evaluated the effect of inhibitors on the replication of MDV in chicken cells.The results showed that the compound b-AP15 significantly non-competitively inhibited the deubiquitinating activity of UL36 at the concentration higher than 5μmol/L(P<0.01).b-AP15 at concentration of 0.25μmol/L and 0.50μmol/L inhibited MDV-induced CPE and MDV replication on CEF cells(P<0.01).Treatment of 2 mg/kg and 5 mg/kg b-AP15 daily by intraperitoneal administration on MDV-challenged chickens significantly suppressed the replication of MDV in chicken T cells(P<0.01).These results suggested that the deubiquitinase inhibitor b-AP15 was able to suppress the replication of MDV in chicken cells.

关 键 词：去泛素化酶UL36 马立克氏病病毒 抑制剂b-AP15 

分 类 号：S831.7[农业科学—畜牧学] S852.65[农业科学—畜牧兽医]