H4N2亚型禽流感病毒(AIV)感染小鼠肺脏细胞因子的表达  

Expression of Cytokines in Lung of Mice (Mus musculus) Infected with H4N2 Subtype Avian influenza virus (AIV)

作　　者：王俊恒[1,2] 石迎[2] 孙海伟[2] 陈鸿军[2] 滕巧泱[2] 刘芹防[2] 李雪松[2] 杨健美[2] 赵宇军[1] 韩克光[1] 李泽君[2] 

机构地区：[1]山西农业大学动物科技学院,太谷030801 [2]中国农业科学院上海兽医研究所,上海200241

出　　处：《农业生物技术学报》2017年第7期1178-1187,共10页Journal of Agricultural Biotechnology

基　　金：国家自然科学基金项目(No.31472206)

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q RT-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。q RT-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。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-γ, IFN-γ in lungs were detected by ELISA and qRT-PCR, respectively. The ELISA results showed that the expressions of IL-1 β, IL-6, IL-10, MCP-1, IFN-γand 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-~/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 oflL-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 oflFN-y 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

关 键 词：禽流感病毒(AIV) H4N2亚型 炎症反应 细胞因子 

分 类 号：S855.3[农业科学—临床兽医学]