[1]许长萌,王志建,王咪,等.猪伪狂犬病毒引起PK-15细胞自噬的研究[J].南京农业大学学报,2018,41(4):701-707.[doi:10.7685/jnau.201801015]
 XU Changmeng,WANG Zhijian,WANG Mi,et al.The study on autophagy induced by pseudorabies virus in PK-15 cells(PRV)[J].Journal of Nanjing Agricultural University,2018,41(4):701-707.[doi:10.7685/jnau.201801015]
点击复制

猪伪狂犬病毒引起PK-15细胞自噬的研究()
分享到:

《南京农业大学学报》[ISSN:1000-2030/CN:32-1148/S]

卷:
41卷
期数:
2018年4期
页码:
701-707
栏目:
出版日期:
2018-07-09

文章信息/Info

Title:
The study on autophagy induced by pseudorabies virus in PK-15 cells(PRV)
作者:
许长萌 王志建 王咪 刘倩玉 王先炜
南京农业大学动物医学院/农业农村部动物疫病诊断与免疫重点开放实验室, 江苏 南京 210095
Author(s):
XU Changmeng WANG Zhijian WANG Mi LIU Qianyu WANG Xianwei
College of Veterinary Medicine/Key Laboratory of Animal Diseases Diagnostic and Immunology, Ministry of Agriculture and Rural Affairs, Nanjing Agricultural University, Nanjing 210095, China
关键词:
自噬猪伪狂犬病毒(PRV)LC3自噬流复制Beclin-1
Keywords:
autophagypseudorabies virus(PRV)LC3autophagic fluxreplicationBeclin-1
分类号:
S852.65
DOI:
10.7685/jnau.201801015
摘要:
[目的]研究猪伪狂犬病毒(PRV)感染猪肾传代细胞系(PK-15)后对细胞自噬的影响,以及细胞自噬在PRV感染复制过程中的作用。[方法]采用Western blot、透射电子显微镜(TEM)、激光共聚焦、siRNA转染等方法研究了PK-15细胞感染PRV后的细胞自噬。利用自噬诱导剂雷帕霉素和自噬抑制剂3-MA分析自噬对病毒复制的影响,再通过沉默自噬相关基因Beclin-1,进一步观察自噬对PRV复制的影响。[结果]PRV感染PK-15细胞后,LC3-Ⅰ显著转化为LC3-Ⅱ,PK-15细胞内自噬小体的数量明显增加。PRV感染增加了LC3阳性斑点的细胞数。同时,变异毒株PRV ZJ01与传统毒株PRV LA诱导的自噬存在差异。自噬的药理学变化试验和siRNA转染结果还显示自噬能够促进PRV的复制。[结论]PRV感染PK-15细胞可以诱导自噬并有利于病毒的复制。
Abstract:
[Objectives]The aim of this research was to investigate the viral inducible-autophagy on PK-15 cell by pseudorabies virus(PRV)and effect of the autophagy on virus replication.[Methods]In this study,Western blot,transmission electron microscopy(TEM),confocal laser scanning microscopy,and transfection were performed to confirm PRV-induced autophagy. The role of autophagy on virus replication was also analyzed by autophagy inducer(rapamycin)and autophagy inhibitor(3-methyladenine,3-MA). By silencing autophagy-related gene Beclin-1, the effect of autophagy on PRV replication was further observed.[Results]Western blot assay showed that LC3-Ⅰ was significantly converted into LC3-Ⅱ in PK-15 cells inoculated with PRV. While the number of auto-phagosome in PK-15 cells infected with PRV were remarkably increased by TEM. In addition,the numbers of LC3-positive puncta per cell in the PRV infected cells were more than in mock cells. These results indicated that PRV infection could induce a complete autophagic flux in PK-15 cells. Furthermore,the aforementioned studies also confirmed that there were differences between autophagy induced by epidemic strains PRV ZJ01 and traditional strains PRV LA. So,the enhancement to PRV replication by autophagy was confirmed in the pharmacological alteration assay and siRNA transfection.[Conclusions]PRV infection can induce autophagy to facilitate virus replication in PK-15 cells.

参考文献/References:

[1] 常凯,朱邵伟,隋昶升. 猪伪狂犬病的研究进展[J]. 湖北畜牧兽医,2013,34(5):67-68. Chang K,Zhu S W,Sui C S. The research development of porcine pseudorabies[J]. Hubei Journal of Animal and Veterinary Sciences,2013,34(5):67-68(in Chinese).
[2] Muller T,Hahn E C,Tottewitz F,et al. Pseudorabies virus in wild swine:a global perspective[J]. Archives of Virology,2011,156(10):1691-1705.
[3] 曾智勇,周莉,汤德元,等. 猪伪狂犬病病毒GZ-Z1株的分离鉴定[J]. 中国兽医科学,2011,41(5):459-463. Zeng Z Y,Zhou L,Tang D Y,et al. Isolation and identification of porcine pseudorabies virus GZ-Z1 strain[J]. Chinese Veterinary Science,2011,41(5):459-463(in Chinese with English abstract).
[4] 华利忠,刘剑锋,冯志新,等. 猪伪狂犬病病毒新流行变异毒株的研究进展[J]. 江苏农业学报,2017,33(2):476-480. Hua L Z,Liu J F,Feng Z X,et al. Progress in novel pseudorabies virus variants prevalent in China[J]. Jiangsu Journal of Agricultural Sciences,2017,33(2):476-480(in Chinese with English abstract).
[5] Freuling C M,Muller T F,Mettenleiter T C. Vaccines against pseudorabies virus(PrV)[J]. Veterinary Microbiology,2017,206:3-9.
[6] 黄锦柳. 猪伪狂犬病新流行毒株的研究进展[J]. 广东畜牧兽医科技,2016,41(6):18-20. Huang J L. Research progress on new prevalent strains of porcine pseudorabies[J]. Guangdong Journal of Animal and Veterinary Science,2016,41(6):18-20(in Chinese with English abstract).
[7] Galluzzi L,Vicencio J M,Kepp O,et al. To die or not to die:that is the autophagic question[J]. Current Molecular Medicine,2008,8(2):78-91.
[8] Deretic V. Autophagy in infection[J]. Current Opinion in Cell Biology,2010,22(2):252-262.
[9] Shintant T,Klionsky D J. Autophagy in health and disease:a double-edged sword[J]. Science,2004,306(5698):990-995.
[10] Yakoub A M,Shukla D. Autophagy stimulation abrogates herpes simplex virus-1 infection[J]. Scientific Reports,2015. DOI:10.1038/srep09730.
[11] Leymarie O,Lepont L,Berlioz-torrernt C. Canonical and non-canonical autophagy in HIV-1 replication cycle[J]. Viruses,2017,9(10):270.
[12] Liu G,Zhong M,Guo C,et al,Autophagy is involved in regulating influenza A virus RNA and protein synthesis associated with both modulation of Hsp90 induction and mTOR/p70S6K signaling pathway[J]. The International Journal of Biochemistry and Cell Biology,2016,72:100-108.
[13] Klionsky D J,Cuervo A M,Seglen P O. Methods for monitoring autophagy from yeast to human[J]. Autophagy,2007,3(3):181-206.
[14] Klionsky D J,Abeliovich H,Agostinis P,et al. Guidelines for the use and interpretation of assays for monitoring autophagy in higher eukaryotes[J]. Autophagy,2008,4(2):151-175.
[15] Mizushima N,Yoshimori T. How to interpret LC3 immunoblotting[J]. Autophagy,2007,3(6):542-545.
[16] Martinet W,Timmermans J P,de Meyer G R. Methods to assess autophagy in situ:transmission electron microscopy versus immunohistochemistry[J]. Methods in Enzymology,2014,543:89-114.
[17] 陈曦,梁文颖,熊静波. 自噬的检测方法及评述[J]. 现代生物医学进展,2012,12(9):1797-1800. Chen X,Liang W Y,Xiong J B. Detection methods of autophagy and the assessments[J]. Progress in Modern Biomedicine,2012,12(9):1797-1800(in Chinese with English abstract).
[18] Wang X,Liu J,Zhen J,et al. Histone deacetylase 4 selectively contributes to podocyte injury in diabetic nephropathy[J]. Kidney International,2014,86(4):712-725.
[19] Yuan K F,Huang C H,John F,et al. Autophagy plays an essential role in the clearance of Pseudomonas aeruginosa by alveolar macrophages[J]. Journal of Cell Science,2012,125:507-515.
[20] Deretic V,Levine B. Autophagy,immunity,and microbial adaptations[J]. Cell Host and Microbe Cell,2009,5(6):527-549.
[21] 叶青,郑民华. 自噬的分子机制与病理生理意义[J]. 国际病理科学与临床杂志,2007,27(4):358-362. Ye Q,Zheng M H. Molecular mechanism of autophagy associated with diseases[J]. International Journal of Pathology and Clinical Medicine,2007,27(4):358-362(in Chinese with English abstract).
[22] Alexander D E,Ward S L,Mizushima N,et al. Analysis of the role of autophagy in replication of herpes simplex virus in cell culture[J]. Journal of Virology,2007,81(22):12128-12134.
[23] English L,Chemali M,Duron J,et al. Autophagy enhances the presentation of endogenous viral antigens on MHC class I molecules during HSV-1 infection[J]. Nature Immunology,2009,10(5):480-487.
[24] Sun M,Hou L,Tang Y D,et al. Pseudorabies virus infection inhibits autophagy in permissive cells in vitro[J]. Scientific Reports,2017,7. DOI:10.1038/strep39964.
[25] Takahashi M N,Jackson W,Laird D T,et al. Varicella-zoster virus infection induces autophagy in both cultured cells and human skin vesicles[J]. Journal of Virology,2009,83(11):5466-5476.
[26] Carpenter J E,Jackson W,Benetti L,et al. Autophagosome formation during varicella-zoster virus infection following endoplasmic reticulum stress and the unfolded protein response[J]. Journal of Virology,2011,85(18):9414-9424.
[27] Buckingham E M,Carpenter J E,Jackson W,et al. Autophagic flux without a block differentiates varicella-zoster virus infection from herpes simplex virus infection[J]. Proc Natl Acad Sci USA,2015,112(1):256-261.
[28] Cong X,Lei J L,Xia S L,et al. Pathogenicity and immunogenicity of a gE/gI/TK gene-deleted pseudorabies virus variant in susceptible animals[J]. Veterinary Microbiology,2016,182:170-177.
[29] Gu Z,Dong J,Wang J,et al. A novel inactivated gE/gI deleted pseudorabies virus(PRV)vaccine completely protects pigs from an emerged variant PRV challenge[J]. Virus Research,2015,195:57-63.

相似文献/References:

[1]李烈川,申明,宁彩波,等.过氧化氢诱导猪卵巢颗粒细胞自噬及其对凋亡的影响[J].南京农业大学学报,2016,39(5):814.[doi:10.7685/jnau.201601001]
 LI Liechuan,SHEN Ming,NING Caibo,et al.Induction of granulosa cell autophagy and effects in apoptosis by hydrogen peroxide in porcine ovaries[J].Journal of Nanjing Agricultural University,2016,39(4):814.[doi:10.7685/jnau.201601001]
[2]韩乐,张国敏,杨花,等.山羊睾丸支持细胞的培养与双荧光自噬报告载体的建立[J].南京农业大学学报,2018,41(1):132.[doi:10.7685/jnau.201612007]
 HAN Le,ZHANG Guomin,YANG Hua,et al.Goat testis sertoli cell culture and autophagy dual-fluorescence reporter system construction[J].Journal of Nanjing Agricultural University,2018,41(4):132.[doi:10.7685/jnau.201612007]
[3]冯楠楠,王冰洁,朱启航,等.玉米赤霉烯酮通过PI3K/Akt/mTOR通路诱导大鼠睾丸支持细胞自噬及对细胞周期分布的影响[J].南京农业大学学报,2018,41(4):708.[doi:10.7685/jnau.201712033]
 FENG Nannan,WANG Bingjie,ZHU Qihang,et al.Effects of zearalenone on the autophagy and cell cycle of sertoli cells by PI3K/Akt/mTOR pathway[J].Journal of Nanjing Agricultural University,2018,41(4):708.[doi:10.7685/jnau.201712033]

备注/Memo

备注/Memo:
收稿日期:2018-01-11。
基金项目:国家重点研究发展项目(2016YFD0500104);中央高校基本科研业务费专项资金(KJQN201919)
作者简介:许长萌,硕士研究生。
通信作者:王先炜,副教授,主要从事动物传染病的教学与研究工作,E-mail:xwwang@njau.edu.cn
更新日期/Last Update: 1900-01-01