您的位置：山东大学 -> 科技期刊社 -> 《山东大学学报（医学版）》

山东大学学报（医学版） ›› 2015, Vol. 53 ›› Issue (12): 16-19.doi: 10.6040/j.issn.1671-7554.0.2015.227

• 基础医学 • 上一篇    下一篇

肠道病毒71型3CD蛋白对细胞的损伤作用

李静, 乔乔, 孙乐乐, 马英伟, 庄志超, 赵丽, 王志玉, 温红玲   

  1. 山东大学公共卫生学院病毒学研究室实验畸形学教育部重点实验室, 山东济南 250012
  • 收稿日期:2015-03-03 出版日期:2015-12-10 发布日期:2015-12-10
  • 通讯作者: 温红玲。E-mail:wenhongling@sdu.edu.cn E-mail:wenhongling@sdu.edu.cn
  • 基金资助:
    国家自然科学基金(81371833);山东省医药卫生科技发展计划(2013WS0211)

Cell injury induced by 3CD protein of Enterovirus 71

LI Jing, QIAO Qiao, SUN Lele, MA Yingwei, ZHUANG Zhichao, ZHAO Li, WANG Zhiyu, WEN Hongling   

  1. Department of Virology, School of Public Health, Key Laboratory for Experimental Teratology of the Ministry of Education, Shandong University, Jinan 250012, Shandong, China
  • Received:2015-03-03 Online:2015-12-10 Published:2015-12-10

PDF (PC)

180

摘要： 目的 研究肠道病毒71型(EV71)3CD蛋白在致病机制中的作用。方法 实验分为对照组、SDLY1组、SDLY107组和SDLY107(1-3CD)组。通过反向遗传技术将弱毒株SDLY1株的3CD蛋白编码区置换到强毒株SDLY107株,形成拯救病毒SDLY107(1-3CD)株。将EV71病毒接种于RD细胞和Vero细胞,于37℃和39.5℃孵育一定时间后,采用MTT试验检测活细胞量,以判断病毒对细胞损伤作用的改变。结果 37℃下,拯救病毒SDLY107(1-3CD)对细胞的损伤作用弱于SDLY107株,但仍强于SDLY1株(P<0.05);39.5℃下,拯救病毒SDLY107(1-3CD)株对细胞的损伤作用较SDLY107株下降(P<0.05),但与SDLY1株的差异无统计学意义(P>0.05)。结论 EV713CD编码区的置换可以改变病毒对细胞的损伤作用,3CD蛋白编码区可能存在影响病毒毒力的位点。这一发现有助于理解EV71的致病机制。

关键词: 肠道病毒71型, 细胞损伤, 3CD蛋白

Abstract: Objective To investigate the roles of 3CD protein on pathogenesis of Enterovirus 71(EV71). Methods The cells were divided into control group, SDLY1 group, SDLY107 group, and SDLY107(1-3CD) group. 3CD region of SDLY107, which was isolated from the severe case, was replaced by that of SDLY1, which was isolated from the mild case, through reverse genetics, and the recombinant virus was named as SDLY107(1-3CD). EV71 strains were incubated at 37℃ and 39.5℃. After some time, the living cells were detected by MTT assay to show cell injury induced by different strains. Results Compared with SDLY107, cell injury induced by SDLY107(1-3CD) decreased at 37℃ and 39.5℃ (both P<0.05). There was statistical difference between cell injury induced by SDLY1 and SDLY107(1-3CD) at 37℃ (P<0.05), while there was no statistical difference between cell injury induced by SDLY1 and SDLY107(1-3CD) at 39.5℃(P>0.05). Conclusion Replacement of EV713CD can change cell injury induced by EV71, and there may be virulence determinants on 3CD region, which may help us understand the pathogenesis of EV71.

Key words: 3CD protein, Cell injury, Enterovirus 71

中图分类号: 

  • R373.2
[1] Mao LX, Wu B, Bao WX, et al. Epidemiology of hand, foot, and mouth disease and genotype characterization of Enterovirus 71 in Jiangsu, China[J]. J Clin Virol, 2010, 49(2):100-104.
[2] Zhang SB, Liao H, Huang CH, et al. Serum types of enterovirus and clinical characteristics of 237 children with hand, foot and mouth disease in Shenzhen[J]. CJCP, 2008, 10(1):38-41.
[3] McMinn P, Lindsay K, Perera D, et al. Phylogenetic analysis of enterovirus 71 strains isolated during linked epidemics in Malaysia, Singapore, and Western Australia[J]. J Virol, 2001, 75(16):7732-7738.
[4] Kehle J, Roth B, Metzger C, et al. Molecular characterization of an Enterovirus 71 causing neurological disease in Germany[J]. J Neurovirol, 2003, 9(1):126-128.
[5] McMinn PC. An overview of the evolution of enterovirus 71 and its clinical and public health significance[J]. FEMS Microbiol Rev, 2002, 26(1):91-107.
[6] Wen HL, Si LY, Yuan XJ, et al. Complete genome sequencing and analysis of six enterovirus 71 strains with different clinical phenotypes[J]. Virol J, 2013, 10(1):115-125.
[7] 袁晓晶,温红玲,高峰,等.肠道病毒71型基因组5'非编码区序列分析[J].山东大学学报:医学版, 2012, 50(9):127-132. YUAN Xiaojing, WEN Hongling, GAO Feng, et al. Sequence analysis for 5' untranslated region of genomes from enterovirus 71[J]. Journal of Shandong University:Health Sciences, 2012, 50(9):127-132.
[8] 温红玲,郝树彬,高峰,等.肠道病毒71型山东临沂分离株全基因组序列分析[J].中华微生物学和免疫学杂志, 2011, 31(7):603-608. WEN Hongling, HAO Shubin, GAO Feng, et al. Sequence analysis for full length genomes of human enterovirus 71 strains isolated in Linyi,Shandong Province[J]. Chinese Journal of Microbiology and Immunology, 2011, 31(7):603-608.
[9] Sun LL, Wang JK, Cui XQ, et al. Association of viral replication capacity with the pathogenicity of enterovirus 71[J]. Virus Res, 2014, 189(5):1-7.
[10] 吴家兵,方益荣,王建军,等.安徽省2008年手足口病流行病学分析[J].安徽预防医学杂志, 2010, 16(2):96-98. WU Jiabing, FANG Yirong, WANG Jianjun, et al. Epidemiological analysis of hand, foot and mouth disease in Anhui Province in 2008[J]. Anhui Journal of Preventive Medicine, 2010, 16(2):96-98.
[11] 刘威龙,韦清,杨桂林,等.深圳5例肠道病毒71型手足口病病毒株全基因组测序分析[J].中华微生物学和免疫学杂志, 2010, 30(5):409-409. LIU Weilong, WEI Qing, YANG Guilin, et al. Complete genome sequencing and analysis of five enterovirus 71 strains in Shenzhen[J]. Chinese Journal of Microbiology and Immunology, 2010, 30(5):409-409.
[12] Barco A, Feduchi E, Carrasco L. Poliovirus protease 3C(pro) kills cells by apoptosis[J]. Virology, 2000, 266(2):352-360.
[13] Tardy-Panit M, Blondel B, Martin A, et al. A mutation in the RNA polymerase of poliovirus type 1 contributes to attenuation in mice[J]. J Virol, 1993, 67(8):4630-4638.
[14] Kung YH, Huang SW, Kuo PH, et al. Introduction of a strong temperature-sensitive phenotype into enterovirus 71 by altering an amino acid of virus 3D polymerase[J]. Virology, 2010, 396(1):1-9.
[15] Clark ME, Lieberman PM, Berk AJ, et al. Direct cleavage of human TATA-binding protein by poliovirus protease 3C in vivo and in vitro[J]. Mol Cell Biol, 1993, 13(2):1232-1237.
[16] Weidman MK, Yalamanchili P, Ng B, et al. Poliovirus 3C protease-mediated degradation of transcriptional activator p53 requires a cellular activity[J]. Virology, 2001, 291(2):260-271.
[17] Falk MM, Grigera PR, Bergmann IE, et al. Foot-and-mouth disease virus protease 3C induces specific proteolytic cleavage of host cell histone H3[J]. J Virol, 1990, 64(2):748-756.
[18] Clark ME, Hämmerle T, Wimmer E, et al. Poliovirus proteinase 3C converts an active form of transcription factor ⅢC to an inactive form:a mechanism for inhibition of host cell polymerase Ⅲ transcription by poliovirus[J]. EMBO J, 1991, 10(10):2941-2947.
[1] 白永娟,庄志超,郝树彬,王莉鸿,李纯,鲁燕,赵丽,王志玉,温红玲. 肠道病毒71型3D重组病毒的拯救[J]. 山东大学学报（医学版）, 2017, 55(8): 7-12.
[2] 马英伟, 孙乐乐, 李静, 乔乔, 庄志超, 赵丽, 于学杰, 王志玉, 温红玲, 孙妍琳. 肠道病毒71型微复制子的构建[J]. 山东大学学报（医学版）, 2015, 53(9): 35-40.
[3] 王小岳, 王道光, 郭继生, 李翠玲, 荆瑞瑞, 吕鑫, 陈新骏, 杨静华. 非标记定量法对乙醇诱导肝损伤的蛋白质组学研究[J]. 山东大学学报（医学版）, 2015, 53(3): 12-16.
[4] 陈更越, 张静, 丁树艳, 马莉莉, 王伟, 田园. HO-1及MIF在子痫前期患者胎盘及脐组织的表达[J]. 山东大学学报（医学版）, 2014, 52(S2): 1-2.
[5] 李敏, 段坤, 赵瑞霞. 2010-2013年济南市历城区414例手足口病病例分析[J]. 山东大学学报（医学版）, 2014, 52(9): 54-57.
[6] 袁晓晶1，温红玲1，高峰2，许洪芝1，司鲁莹1，王东旭1，宋艳艳1，赵丽1，王志玉1. 肠道病毒71型基因组5′非编码区序列分析[J]. 山东大学学报(医学版), 2012, 50(9): 127-.
[7] 王东旭1，温红玲1，董华1，孙成玺1，司鲁莹1，袁晓晶1，褚福禄1，赵丽1，王志玉1, 2. 不同免疫方案制备肠道病毒71型抗血清的免疫效果比较[J]. 山东大学学报(医学版), 2012, 50(3): 129-.
[8] 李志会，岳盈盈，宋楠楠，李鹏，孟红. 制备肠道病毒71型黏膜疫苗的初步探索[J]. 山东大学学报(医学版), 2012, 50(2): 25-.
[9] 李鹏1,张颖2,岳盈盈1,宋楠楠1,李志会1,盖中涛2,孟红1. 肠道病毒71型山东分离株全基因组序列分析[J]. 山东大学学报(医学版), 2011, 49(10): 113-117.
[10] 薛宁，王纪文,张洪伟. 大剂量甲泼尼龙冲击治疗重症手足口病的意义[J]. 山东大学学报(医学版), 2010, 48(3): 106-.
[11] 秦文,王立祥,曾季平,陈蔚文,张亚伦 . 甲基环戊二烯羰基锰对PC-3M细胞损伤作用[J]. 山东大学学报(医学版), 2007, 45(7): 655-657.
Viewed
Full text


Abstract

Cited
  Shared   
  Discussed   
No Suggested Reading articles found!
版权所有 © 2018 《山东大学学报(医学版)》编辑部　
地址:山东大学科技期刊社（济南市历城区山大南路27号山东大学中心校区明德楼B座721室） 电话: 0531-88366918 E-mail: xbyxb@sdu.edu.cn
本系统由北京玛格泰克科技发展有限公司设计开发