甲型流感病毒重组血凝素1诱导小鼠气管上皮细胞产生β-防御素和γ-干扰素  

Influenza A virus recombinant hemagglutinin 1 induces the production of beta-defensin and interferon-gamma in mouse tracheal epithelial cells

作　　者：雷琪 赵兵兵 罗红 陈强 江滟 Lei Qi;Zhao Bingbing;Luo Hong;Chen Qiang;Jiang Yan(Clinical Laboratory Center,Affiliated Hospital of Guizhou Medical University,Guiyang 550028,Guizhou Province,China;Department of Clinical Microbiology and Immunology,School of Medical Laboratory Medicine,Guizhou Medical University,Guiyang 550025,Guizhou Province,China;Laboratory Animal Center,Guizhou Medical University,Guiyang 550000,Guizhou Province,China;The Second People's Hospital of Guizhou Province,Guiyang 550000,Guizhou Province,China)

机构地区：[1]贵州医科大学附属医院临床检验中心,贵州省贵阳市550028 [2]贵州医科大学医学检验学院临床微生物与免疫学教研室,贵州省贵阳市550025 [3]贵州医科大学实验动物中心,贵州省贵阳市550000 [4]贵州省第二人民医院,贵州省贵阳市550000

出　　处：《中国组织工程研究》2025年第32期6905-6912,共8页Chinese Journal of Tissue Engineering Research

基　　金：国家自然科学基金(81260249),项目负责人:江滟;贵州省科技计划项目(黔科合基础-ZK[2022]一般228),项目负责人:陈强;贵州医科大学慢性疾病标志物研究重点实验室项目(校重点实验室[2024]fy004号)~~。

摘　　要：背景:β-防御素具有抗甲型流感病毒活性,并能抑制甲型流感病毒感染引起的细胞内一系列炎症反应。关于甲型流感病毒表面蛋白血凝素1单独作用于原代小鼠气管上皮细胞后,是否能够诱导分泌β-防御素和γ-干扰素还未见研究报道。目的:探讨重组血凝素1诱导小鼠气管上皮细胞产生β-防御素和γ-干扰素的水平。方法:将原代小鼠气管上皮细胞分为6组:空白对照组、重组血凝素1组(200 ng/mL)、重组血凝素1+甲型流感病毒组、甲型流感病毒组(2×TCID50)、灭活甲型流感病毒组、重组血凝素1+灭活甲型流感病毒组。各组分别处理小鼠气管上皮细胞4,8,24 h后,苏木精-伊染色行病理学观察;qRT-PCR法检测细胞中β-防御素2,3,4及γ-干扰素的mRNA表达水平;ELISA检测细胞中β-防御素2,3,4的水平;Western blot检测γ-干扰素的蛋白表达水平。结果与结论:①重组血凝素1单独作用或与甲型流感病毒联合作用可致气管上皮细胞出现不同程度的病理变化,可观察到细胞出现空泡、核固缩以及细胞融合等现象;②与空白对照组相比,在重组血凝素1单独存在或与甲型流感病毒或灭活甲型流感病毒共同存在的情况下,可诱导小鼠气管上皮细胞分泌β-防御素2、β-防御素3、β-防御素4及γ-干扰素(P<0.05);③结果表明,重组血凝素1单独或联合甲型流感病毒均能诱导小鼠气管上皮细胞产生β-防御素2-4和γ-干扰素。BACKGROUND:β-Defensin has the ability against influenza A virus and inhibits a series of inflammatory responses induced by influenza A virus infection within cells.There have been no reports on whether hemagglutinin 1 from influenza A virus can induce the secretion of mouseβ-defensin and interferon-γwhen acting in mouse tracheal epithelial cells.OBJECTIVE:To investigate the effect of recombinant hemagglutinin 1 on the production levels of mouseβ-defensin and interferon-γin mouse tracheal epithelial cells.METHODS:Primary mouse tracheal epithelial cell were divided into six groups:blank control group(Control),recombinant hemagglutinin 1 group(200 ng/mL),recombinant hemagglutinin 1+influenza A virus group,influenza A virus group(2×TCID50),recombinant hemagglutinin 1+inactivated influenza A virus group,and inactivated influenza A virus(I)group.After the mouse tracheal epithelial cells in each group were treated for 4,8,or 24 hours,hematoxylin-eosin staining was used for pathological observation.The mRNA levels of mouseβ-defensin 2,3,and 4,and interferon-γwere detected by qRT-PCR.The protein levels of mouseβ-defensin 2,3,and 4 were measured by enzyme-linked immunosorbent assay,and the protein expression of interferon-γwas detected by western blot.RESULTS AND CONCLUSION:(1)The recombinant hemagglutinin 1 acting alone or in combination with influenza A virus could cause different pathological changes in tracheal epithelial cells.Phenomena such as vacuolation,nuclear pyknosis and cell fusion could be observed in the cells.(2)Compared with the control group,recombinant hemagglutinin 1 alone or in combination with influenza A virus or inactivated influenza A virus significantly induced the production of mouseβ-defensin 2,3,and 4(P<0.05)and interferon-γin mouse tracheal epithelial cells(P<0.05).These results indicate that recombinant hemagglutinin 1 alone or in combination with influenza A virus can induce the production of mouseβ-defensin 2,3,and 4 and interferon-γin mouse tracheal epithelial cells.

关 键 词：甲型流感病毒 重组血凝素1 Β-防御素 Γ-干扰素 气管上皮细胞 小鼠 

分 类 号：R459.9[医药卫生—治疗学] R318[医药卫生—临床医学] R446