鸭甲肝病毒3型在人工感染雏鸭体内分布与诱导肝病变及细胞因子表达的关联性分析

doi:10.11843/j.issn.0366-6964.2018.01.019

畜牧兽医学报 ›› 2018, Vol. 49 ›› Issue (1): 164-172.doi: 10.11843/j.issn.0366-6964.2018.01.019

鸭甲肝病毒3型在人工感染雏鸭体内分布与诱导肝病变及细胞因子表达的关联性分析

朱玉东^1,2, 汪铭书^1,2,3, 程安春^1,2,3*, 朱德康^1,2,3, 贾仁勇^1,2,3, 刘马峰^1,2,3, 陈舜^1,2,3, 赵新新^1,2,3, 杨乔^1,2,3, 吴英^1,2,3, 陈孝跃^1,3

1. 四川农业大学预防兽医研究所, 成都 611130;
2. 四川农业大学动物疫病与人类健康四川省重点实验室, 成都 611130;
3. 四川农业大学动物医学院禽病防治研究中心, 成都 611130

收稿日期:2017-08-17 出版日期:2018-01-23 发布日期:2018-01-23
通讯作者: 程安春,教授,主要从事动物传染病学的教学和科研工作,E-mail:chenganchun@vip.163.com
作者简介:朱玉东(1989-),男,山东费县人,硕士生,主要从事动物传染病研究,E-mail:597295631@qq.com
基金资助:
“十三五”国家重点研发计划项目（2017YFD0500800）；国家现代农业产业技术体系专项（CARS-42-17）；动物疫病与人类健康四川省重点实验室专项（2016JPT0004）

Distribution of Duck Hepatitis A Virus Type 3 in Artificial Infected Ducklings and Analysis of the Relationship between Histopathological Changes and Expression of Cytokines in the Livers

ZHU Yu-dong^1,2, WANG Ming-shu^1,2,3, CHENG An-chun^1,2,3*, ZHU De-kang^1,2,3, JIA Ren-yong^1,2,3, LIU Ma-feng^1,2,3, CHEN Shun^1,2,3, ZHAO Xin-xin^1,2,3, YANG Qiao^1,2,3, WU Ying^1,2,3, CHEN Xiao-yue^1,3

1. Institute of Preventive Veterinary Medicine, Sichuan Agricultural University, Chengdu 611130, China;
2. Sichuan Key Laboratory of Animal Diseases and Human Health, Sichuan Agricultural University, Chengdu 611130, China;
3. College of Veterinary Medicine, Sichuan Agricultural University, Chengdu 611130, China

Received:2017-08-17 Online:2018-01-23 Published:2018-01-23

摘要/Abstract

摘要：

探究鸭甲肝病毒3型（DHAV-3）在感染7日龄雏鸭体内动态分布规律与致肝病变和诱导IFN及促炎因子表达之间的关联。以DHAV-3强毒感染7日龄雏鸭，于感染后1、3、6、9、12、24、48、72和96 h采集雏鸭的血液、肝、脾、肾、脑、胰、肺、胸腺、法氏囊、哈德氏腺和十二指肠共11种组织样品，以一步法TaqMan探针实时荧光定量RT-PCR检测病毒含量变化、染料法实时荧光定量RT-PCR检测肝中抗病毒细胞因子（IFN-α、IFN-β和IFN-γ）及促炎因子（IL-1β、IL-2和IL-6）在转录水平的变化，用光学显微镜对肝组织病理学变化进行观察，分析这些变化之间的联系。结果表明：DHAV-3感染1 h后即在所有的样品中检测到。组织器官中病毒含量，血液和胰分别在感染后12和96 h达到峰值，其余器官均在感染24~48 h后达到峰值。病毒含量较高的前三位器官是肝、脾和肾（分别为10^11.15、10^10.37和10^10.30 copies·g^-1）。肝组织病理学变化在感染3~12 h后以空泡变性为主、24~48 h以坏死为主。肝中上述6种细胞因子转录量除IL-1β在12 h达到最高外，其余均在感染后24~48 h达到最高，感染48 h后，其转录量变化随肝中病毒含量下降而降低，并与肝病变严重程度呈正相关。DHAV-3在雏鸭体内具有广泛的组织嗜性，肝、脾、肾是病毒攻击的主要靶器官，肝中细胞因子极有可能在抑制病毒增殖和修复组织损伤过程中发挥重要作用。

Abstract:

This experiment was conducted to explore the dynamic distribution of type 3 duck hepatitis A virus (DHAV-3) and the relationship between the induction of histopathological, antiviral cytokines and pro-inflammatory cytokines in 7-day-old ducklings. One week old ducklings were infected with DHAV-3. At 1, 3, 6, 9, 12, 24, 48, 72 and 96 h after infection, 11 tissue samples of blood, liver, spleen, pancreas, kidney, lung, brain, thymus, bursa of Fabricius, Harderian glands and duodenum were collected randomly from three ducklings, and then detected. The virus load was detected by TaqMan real-time one-step RT-PCR and the pathological damage changes of liver was observed by optical microscope, the transcription of anti-viral cytokines (IFN-α, IFN-β and IFN-γ) and pro-inflammatory cytokines (IL-1β, IL-2 and IL-6) in liver were detected by real-time RT-PCR at different time points. The links between these changes were analyzed. The results showed that the virus can be detected in all tissues of 1-week-old ducklings in 1 h after infection. The highest load of virus in tissues was at 24-48 h after infection, except that the highest of blood and pancreas were at 12 h and 96 h after infection respectively. The three organs with the highest viral load were liver, spleen and kidney (10^11.15, 10^10.37 and 10^10.30 copies·g^-1). Liver main histopathological changes were vacuoles at 3-12 h, and 24-48 h for the necrotic changes. The transcription of 6 cytokines in the liver reached the highest level at 24-48 h (except for IL-1β at 12 h) after infection. After 48 h of infection, the transcription level of the cytokines decreased with the decrease of the virus content in the liver and was positively correlated with the severity of liver disease. DHAV-3 has extensive tissue tropism in ducklings. Liver, spleen and kidney are the main target organ of virus attack. The cytokines in the liver are highly likely to play an important role in inhibiting viral proliferation and repairing tissue damage.

中图分类号:

S852.659.6

朱玉东, 汪铭书, 程安春, 朱德康, 贾仁勇, 刘马峰, 陈舜, 赵新新, 杨乔, 吴英, 陈孝跃. 鸭甲肝病毒3型在人工感染雏鸭体内分布与诱导肝病变及细胞因子表达的关联性分析[J]. 畜牧兽医学报, 2018, 49(1): 164-172.

ZHU Yu-dong, WANG Ming-shu, CHENG An-chun, ZHU De-kang, JIA Ren-yong, LIU Ma-feng, CHEN Shun, ZHAO Xin-xin, YANG Qiao, WU Ying, CHEN Xiao-yue. Distribution of Duck Hepatitis A Virus Type 3 in Artificial Infected Ducklings and Analysis of the Relationship between Histopathological Changes and Expression of Cytokines in the Livers[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2018, 49(1): 164-172.

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鸭甲肝病毒3型在人工感染雏鸭体内分布与诱导肝病变及细胞因子表达的关联性分析

Distribution of Duck Hepatitis A Virus Type 3 in Artificial Infected Ducklings and Analysis of the Relationship between Histopathological Changes and Expression of Cytokines in the Livers

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