微管对狂犬病病毒胞内感染的影响

doi:10.14088/j.cnki.issn0439-8114.2019.10.028

湖北农业科学 ›› 2019, Vol. 58 ›› Issue (10): 121-124.doi: 10.14088/j.cnki.issn0439-8114.2019.10.028

微管对狂犬病病毒胞内感染的影响

高洁^a, 赵铭昕^a,b, 刘恩华^a,b, 赵维荣^a,b, 段铭^a, 关振宏^a, 张茂林^a, 郭艺迪^a

吉林大学,a.人兽共患病研究所/人兽共患病研究教育部重点实验室; b.动物医学学院,长春 130062

收稿日期:2019-03-01 出版日期:2019-05-25 发布日期:2019-12-03
通讯作者: 郭艺迪（1988-）,女,黑龙江齐齐哈尔人,博士,主要从事病毒学研究,（电子信箱）guoyd@jlu.edu.cn。
作者简介:高洁（1993-）,女,河北石家庄人,硕士,主要从事兽医公共卫生研究,（电话）18844094249（电子信箱）1908053854@qq.com。
基金资助:
国家重要研发计划项目（216YFD0500402）; 国家自然科学基金项目（31702238; 31472208）

Effects of microtubules on RABV intracellular infection

GAO Jie^a, ZHAO Ming-xin^a,b, LIU En-hua^a,b, ZHAO Wei-rong^a,b, DUAN Ming^a, GUAN Zhen-hong^a, ZHANG Mao-lin^a, GUO Yi-di^a

a.Key Laboratory of Zoonosis,Ministry of Education/Institute of Zoonosis; b.College of Veterinary Medicine,Jilin University,Changchun 130062,China

Received:2019-03-01 Online:2019-05-25 Published:2019-12-03

摘要/Abstract

摘要： 狂犬病病毒（Rabies virus,RABV）依赖宿主细胞完成其感染周期,且在胞浆内复制。微管作为一种细胞骨架,是介导胞内物质运输的重要通道。为探究狂犬病病毒胞内感染是否依赖于细胞骨架——微管,利用小鼠神经母细胞瘤细胞（N2a）,接种实验室标准攻击毒株CVS-11,通过诺考达唑（Nocodazole）抑制剂破坏微管形成,采用实时荧光定量PCR方法、Western Blot方法、免疫荧光方法检测微管对于RABV感染量的影响,并采用TCID₅₀法测定微管对RABV病毒滴度的影响。结果表明,Nocodazole通过抑制微管的形成,使RABV N基因水平降低25%、N蛋白水平降低50%,免疫荧光检测到病毒感染率降低70%,上清液中病毒滴度由10^6.5TCID₅₀/mL降低至10^4.5TCID₅₀/mL,说明狂犬病病毒胞内感染依赖于细胞骨架-微管的参与。该结果为进一步研究狂犬病病毒胞内运输及其复制机制的研究提供了理论依据。

关键词: 狂犬病病毒, 微管, 诺考达唑, 胞内感染

Abstract: Rabies virus （RABV） is entirely accomplished its infection in cells cytoplasmic. Cytoskeletal microtubules mediates cargoes to move throughout the cytoplasm. In this study, we investigated the modulatory effect of a microtubule-inhibitor, Nocodazole, on RABV infection in mouse Neuro-2a cells （N2a）. The N2a cells viability was analyzed by MTT. In cells treated with Nocodazole, then infected with laboratory challenge virus standard CVS-11. mRNA level and protein level of RABV were respectively reduced about 25% and 50% by real-time PCR and Western Blot. Significantly, viral infection rate was obviously decent 70% by immunofluorescence and the virus titer was decreased from 10^6.5TCID₅₀/mL to 10^4.5TCID₅₀/mL. These results indicated that RABV undergo intracellular transport mediated by cytoskeleton-microtubule. These results provide a theoretical basis for further study on the intracellular trafficking of RABV and its replication mechanism.

Key words: rabies virus, microtubule, Nocodazole, intracellular infection

中图分类号:

S851

高洁, 赵铭昕, 刘恩华, 赵维荣, 段铭, 关振宏, 张茂林, 郭艺迪. 微管对狂犬病病毒胞内感染的影响[J]. 湖北农业科学, 2019, 58(10): 121-124.

GAO Jie, ZHAO Ming-xin, LIU En-hua, ZHAO Wei-rong, DUAN Ming, GUAN Zhen-hong, ZHANG Mao-lin, GUO Yi-di. Effects of microtubules on RABV intracellular infection[J]. HUBEI AGRICULTURAL SCIENCES, 2019, 58(10): 121-124.

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微管对狂犬病病毒胞内感染的影响

Effects of microtubules on RABV intracellular infection

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