蛋白激酶C抑制剂及激活剂对鸭肠炎病毒增殖的影响

doi:10.16431/j.cnki.1671-7236.2018.07.034

摘要/Abstract

摘要：

为进一步探究鸭肠炎病毒（duck enteritis virus，DEV）的致病机理，试验采用鸭蛋白激酶C （protein kinase C，PKC）的抑制剂星形孢菌素（staurosporine，SP）和激活剂佛波醇（phorbol-12-myristate-13-acetate，PMA）研究PKC/PKCI能否对DEV的增殖产生影响，为阐述DEV致病机理提供新的思路。用SP/PMA处理正常鸭胚成纤维细胞（duck embryo fibroblast，DEF）后，实时荧光定量PCR方法检测不同时间段PKC/PKCI基因表达；用DEV病毒液感染SP/PMA处理的DEF后，收集不同时间段培养物，以Reed-Muench法计算DEV半数组织培养感染量（TCID₅₀）值，实时荧光定量PCR方法检测分析DEV NP基因转录水平。结果显示，SP处理对宿主细胞PKC/PKCI基因表达水平无显著影响（P>0.05）；而PMA处理可极显著提高宿主细胞PKC基因表达（P<0.01），但对PKCI基因表达水平无显著影响（P>0.05）；SP/PMA处理对DEV复制能力影响显著（P<0.05；P<0.01），且在感染前期可显著或极显著降低DEV NP基因的转录水平（P<0.05；P<0.01）。综上所述，SP和PMA对PKC、PKCI基因表达水平的影响效果不同，且都能有效抑制DEV增殖，本试验结果可为DEV的防控及其致病机理的研究提供基础资料。

关键词: 鸭肠炎病毒; 蛋白激酶C (PKC); PKC抑制剂; PKC激活剂; 增殖

Abstract:

To identify the pathogenesis of duck enteritis virus (DEV),inhibitor and activator of PKC (SP and PMA) were used to research that whether PKC/PKCI could affect the proliferation of DEV,trying to provide some new ideas for understanding the mechanism of DEV infection.Normal duck fibroblasts(DEF) were treated with SP or PMA,and Real-time quantitative PCR method was used to measure the expression levels of PKC and PKCI genes at different time.The DEV was used to infect DEF which treated with SP or PMA,then the cell cultures were collected at different time,and the TCID₅₀ and expression levels of DEV NP gene were measured by Reed-Muench and Real-time quantitative PCR,respectively.The results showed that there was no significant effect of SP treatment on PKC/PKCI genes expression levels in DEF (P>0.05),while the PMA treatment could extremely significantly increased the PKC gene expression (P<0.01),but had no significant impact on PKCI gene (P>0.05);SP/PMA processing could significant or extremely significant affect the ability of DEV replication (P<0.05;P<0.01),and the expression level of DEV NP gene was significantly or extremely significant decreased at the early stage of infection (P<0.05;P<0.01).The above results indicated that the effects of SP and PMA on PKC/PKCI were different,while both of them could effectively inhibit the proliferation of DEV.These results could provide the basis for DEV prevention and its pathogenesis studying.

Key words: duck enteritis virus (DEV); protein kinase C (PKC); PKC inhibitor; PKC activator; proliferation

中图分类号:

S853.65

贺欣薇, 苟万里, 张明洋, 周碧君, 程振涛, 文明. 蛋白激酶C抑制剂及激活剂对鸭肠炎病毒增殖的影响[J]. 《中国畜牧兽医》, 2018, 45(7): 2001-2007.

HE Xinwei, GOU Wanli, ZHANG Mingyang, ZHOU Bijun, CHENG Zhentao, WEN Ming. Effects of Protein Kinase C Inhibitor and Activator on Proliferation of Duck Enteritis Virus[J]. , 2018, 45(7): 2001-2007.

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