[1]梁荣,宋海鑫,黄佳玲,等.PEDV流行毒株JS2013诱导Vero细胞凋亡[J].南京农业大学学报,2021,44(3):514-520.[doi:10.7685/jnau.202010010]
 LIANG Rong,SONG Haixin,HUANG Jialing,et al.PEDV epidemic strain JS2013 induced apoptosis of Vero cells[J].Journal of Nanjing Agricultural University,2021,44(3):514-520.[doi:10.7685/jnau.202010010]
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PEDV流行毒株JS2013诱导Vero细胞凋亡()
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《南京农业大学学报》[ISSN:1000-2030/CN:32-1148/S]

卷:
44卷
期数:
2021年3期
页码:
514-520
栏目:
动物科学
出版日期:
2021-05-10

文章信息/Info

Title:
PEDV epidemic strain JS2013 induced apoptosis of Vero cells
作者:
梁荣12 宋海鑫12 黄佳玲12 费荣梅1 张金秋23
1. 南京农业大学动物医学院, 江苏 南京 210095;
2. 江苏省农业科学院动物免疫工程研究所, 江苏 南京 210014;
3. 江苏省动物重要疫病与人兽共患病防控协同创新中心, 江苏 扬州 225009
Author(s):
LIANG Rong12 SONG Haixin12 HUANG Jialing12 FEI Rongmei1 ZHANG Jinqiu23
1. College of Veterinary Medicine, Nanjing Agricultural University, Nanjing 210095, China;
2. Institute of Veterinary Immunology&Engineering, Jiangsu Academy of Agricultural Sciences, Nanjing 210014, China;
3. Jiangsu Co-Innovation Center for the Prevention and Control of Important Animal Infectious Disease and Zoonose, Yangzhou 225009, China
关键词:
猪流行性腹泻病毒流行毒株细胞凋亡Caspase
Keywords:
porcine epidemic diarrhea virusepidemic straincell apoptosisCaspase
分类号:
S852.4
DOI:
10.7685/jnau.202010010
摘要:
[目的] 本试验旨在研究猪流行性腹泻病毒(porcine epidemic diarrhea virus,PEDV)流行毒株JS2013诱导Vero细胞凋亡的相关机制。[方法] 用添加5 μg·mL-1胰酶的Vero细胞成功扩增PEDV JS2013毒株后,采用倒置显微镜、间接免疫荧光、流式细胞术、Hoechst染色、细胞活性计数(CCK-8)以及Western blot等方法,检测Vero细胞感染JS2013后产生的病变、凋亡细胞比例、细胞活性、凋亡相关蛋白的表达等。[结果] PEDV JS2013感染Vero细胞后6 h即有少量病毒进入细胞,24 h后出现典型合胞体病变,PEDV JS2013诱导Vero细胞凋亡呈剂量依赖性,并且显著抑制Vero细胞活性。Hoechst染色后可以观察到明显的凋亡小体。Western blot结果表明,感染PEDV JS2013 48 h后激活Caspase 8和Caspase 9,60 h左右激活Caspase 3。[结论] PEDV流行毒株JS2013可以诱导Vero细胞凋亡。
Abstract:
[Objectives] The study aimed to explore the mechanism of apoptosis in Vero cells induced by porcine epidemic diarrhea virus(PEDV) epidemic strain JS2013. [Methods] PEDV JS2013 was successfully amplified by Vero cells containing 5 μg·mL-1 trypsin. Inverted microscope,indirect immunofluorescence,flow cytometry,Hoechst staining,CCK-8 and Western blot were used to detect the cytopathic effect,the proportion of apoptotic cells,the cell activity and the expression of related apoptotic proteins of Vero cells after infection with JS2013. [Results] A small amount of virus entered the cells at 6 h after PEDV JS2013 infection and typical syncytium lesions appeared at 24 h after infection. PEDV JS2013 induced apoptosis in a dose-dependent manner and significantly inhibited the activity of Vero cells. Hoechst staining showed that obvious apoptotic bodies could be observed in Vero cells. Western blot results showed that Caspase 8 and Caspase 9 were activated at 48 h,and Caspase 3 was activated at 60 h after infection. [Conclusions] PEDV epidemic strain JS2013 can induce apoptosis of Vero cells.

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备注/Memo

备注/Memo:
收稿日期:2020-10-09。
基金项目:国家自然科学基金项目(31772701);江苏省农业科技自主创新资金[CX(17)3027];家畜疫病病原生物学国家重点实验室(中国农业科学院兰州兽医研究所)开放基金课题(SKLVEB2020KFKT007)
作者简介:梁荣,硕士研究生。
通信作者:张金秋,副研究员,从事天然免疫与畜禽疫病防控研究,E-mail:jqzh03@126.com。
更新日期/Last Update: 1900-01-01