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2025年8月11日 星期一
  2017, Vol. 25 Issue (7): 1178-1187    
  专题:不同亚型低致病性禽流感病毒生物学特性研究 本期目录 | 过刊浏览 | 高级检索 |
H4N2亚型禽流感病毒(AIV)感染小鼠肺脏细胞因子的表达
王俊恒1,石迎2,孙海伟3,陈鸿军4,滕巧泱4,刘芹防4,李雪松4,杨建美4,赵宇军1,韩克光1,李泽君4
1. 山西农业大学
2.
3. 上海兽医研究所
4. 中国农业科学院上海兽医研究所
Expression of Cytokines in Lung of Mice (Mus musculus) Infected with H4N2 Subtype Avian influenza virus (AIV)
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摘要 H4N2亚型禽流感病毒(Avian influenza virus, AIV)对哺乳动物健康具有潜在威胁。为探究炎症相关细胞因子在H4N2亚型AIV致病过程中的作用,将106鸡胚半数感染量(fifty percent embryo infection dose, EID50)的P20和P50病毒经鼻腔感染BALB/C小鼠(Mus musculus),于感染后1、3、5 d,利用酶联免疫吸附测定(enzyme-linked immuno sorbent assay, ELISA)和qRT-PCR对肺脏中抑炎细胞因子(白细胞介素2(interleukin-2, IL-2), IL-4, IL-10)、促炎细胞因子(IL-1β, IL-6, 肿瘤坏死因子α(tumour necrosis factor-α, TNF-α))、单核细胞趋化蛋白1 (mononuclear chemokine protein 1, MCP-1)及干扰素α(interferon-α, IFN-α)、IFN-β、IFN-γ分别进行检测。ELISA检测结果发现,小鼠感染病毒后,IL-1β、IL-6、IL-10、MCP-1、IFN-γ和TNF-α均出现不同程度的上调,而IL-2、IL-4、IFN-α/β均出现下调。P50组中IL-6在攻毒后第3天和第5天表达量极其显著高于P20组(P<0.001),IL-10表达变化不明显,但在攻毒后第5天表达量极其显著高于P20组(P<0.001),P20组中MCP-1攻毒后第3天和第5天的表达量均极其显著低于P50组(P<0.001),P50组的IFN-γ在第1、3、5天表达量极显著(P<0.01)或极其显著(P<0.001)高于P20组。qRT-PCR检测结果发现,P50组中IL-6的表达水平在第3天和第5天极显著高于P20组(P<0.01),而P20组与P50组的MCP-1和IFN-γ在转录水平没有统计学差异。为了验证IL-6的促炎症作用,在小鼠鼻腔感染P50病毒后,静脉注射抗IL-6中和抗体,结果表明,各组中IL-10、MCP-1和IFN-γ的表达水平均未表现统计学差异,且没有减轻炎症反应,未对小鼠产生保护作用。本研究表明H4N2亚型AIV对小鼠的致病性与炎症反应相关,但降低IL-6不能减轻炎症反应。研究结果为开发可以用于预防和控制A型流感病毒感染的候选药物提供依据。
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王俊恒
石迎
孙海伟
陈鸿军
滕巧泱
刘芹防
李雪松
杨建美
赵宇军
韩克光
李泽君
关键词 禽流感病毒(AIV)H4N2亚型炎症反应细胞因子    
Abstract:H4N2 subtype Avian influenza virus (AIV) has potential threat to the health of mammalian. To investigate the roles of inflammation-associated cytokines in pathopoiesis development of H4N2 subtype AIV, mice (Mus musculus) were inoculated intranasally with 106 fifty percent embryo infection dose (EID50) of P20 and P50 strains. At 1, 3 and 5 days after infection, the expression of anti-inflammatory cytokines (interleukin-2 (IL-2), IL-4 and IL-10), pro-inflammation cytokines (IL-1β, IL-6 and tumour necrosis factor-α (TNF-α)), monocyte chemoattractant protein-1 (MCP-1) and interferon-α (IFN-α), IFN-β, IFN-γ in lungs were detected by ELISA and qRT-PCR, respectively. The ELISA results showed that the expressions of IL-1β, IL-6, IL-10, MCP-1, IFN-γ and TNF-α were up-regulated, while the expressions of IL-2, IL-4 and IFN-α/β were down-regulated after the infection of both strains. Compared with P20 group, the expression level of IL-6 in P50 group very extremely significantly increased at 3 and 5 days post infection (P<0.001), but the expression level of IL-10 was very extremely significantly higher than that in P20 group at 5 days post infection (P<0.001). MCP-1 expression in P20 group was very extremely significantly lower than that in P50 group at 3 and 5 days post infection (P<0.001). IFN-γ expression in P50 group was extremely significantly (P<0.01) or very extremely significantly (P<0.001) higher than that in P20 group at 1, 3 and 5 days post infection. The mRNA expression of IL-6 in P50 group showed significant increase compared to P20 group at 3 and 5 days post infection (P<0.01). But the mRNA expression of IFN-γ and MCP-1 in both P20 and P50 groups had no significant changes post infection. To validate the role of IL-6 in pro-inflammation, mice were inoculated intravenously with anti-IL-6 neutralizing antibody after inoculated intranasally with P50 virus. The results showed that the expression levels of IL-6, MCP-1 and IFN-γ in each group were not statistically different, and inflammatory response was not alleviated, and it did not produce a protective effect on mice. This study showed that H4N2 subtype AIV was associated with inflammatory response in mice, but reduction of IL-6 could not alleviate the inflammatory response. The results provide a basis for the development of candidate drugs that can be used to prevent and control influenza A virus infection.
Key wordsAvian influenza virus (AIV)    H4N2 subtype    Inflammatory    Cytokines
收稿日期: 2017-02-21      出版日期: 2017-06-16
基金资助:自然科学基金(面上)项目“促进禽流感病毒变异的因素”
通讯作者: 李泽君     E-mail: lizejun@shvri.ac.cn
引用本文:   
王俊恒 石迎 孙海伟 陈鸿军 滕巧泱 刘芹防 李雪松 杨建美 赵宇军 韩克光 李泽君. H4N2亚型禽流感病毒(AIV)感染小鼠肺脏细胞因子的表达[J]. , 2017, 25(7): 1178-1187.
链接本文:  
http://journal05.magtech.org.cn/Jwk_ny/CN/     或     http://journal05.magtech.org.cn/Jwk_ny/CN/Y2017/V25/I7/1178
 
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