中国组织工程研究

中国组织工程研究 ›› 2025, Vol. 29 ›› Issue (32): 6905-6912.doi: 10.12307/2025.922

• 组织构建细胞学实验 cytology experiments in tissue construction • 上一篇    下一篇

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

雷  琪1，2，赵兵兵2，罗  红3，陈  强4，江  滟1，2   

  1. 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
  • 收稿日期:2024-09-24 接受日期:2024-11-25 出版日期:2025-11-18 发布日期:2025-04-26
  • 通讯作者: 江滟，博士，教授，研究生导师，贵州医科大学附属医院临床检验中心，贵州省贵阳市 550028；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
  • 作者简介:雷琪，女，1995年生，贵州省桐梓县人，汉族，贵州医科大学在读硕士，主管检验师，主要从事医学病毒学研究。
  • 基金资助:
    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)

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

Lei Qi1, 2, Zhao Bingbing2, Luo Hong3, Chen Qiang4, Jiang Yan1, 2   

  1. 1Clinical Laboratory Center, Affiliated Hospital of Guizhou Medical University, Guiyang 550028, Guizhou Province, China; 2Department of Clinical Microbiology and Immunology, School of Medical Laboratory Medicine, Guizhou Medical University, Guiyang 550025, Guizhou Province, China; 3Laboratory Animal Center, Guizhou Medical University, Guiyang 550000, Guizhou Province, China; 4The Second People's Hospital of Guizhou Province, Guiyang 550000, Guizhou Province, China
  • Received:2024-09-24 Accepted:2024-11-25 Online:2025-11-18 Published:2025-04-26
  • Contact: Jiang Yan, MD, Professor, Master’s supervisor, 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
  • About author:Lei Qi, Master candidate, Laboratorian in charge, 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
  • Supported by:
    National Natural Science Foundation of China, No. 81260249 (to JY); Guizhou Provincial Science and Technology Program, No. ZK[2022] General 228 (to CQ); a grant for the Key Laboratory of Chronic Disease Marker Research, Guizhou Medical University, No. [2024]fy004

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摘要：


文题释义：
β-防御素(β-defensin，BD)：是一类重要的抗菌肽，广泛存在于哺乳动物体内，具有广谱的抗菌活性，能够有效对抗细菌、真菌甚至一些病毒。β-防御素在流感防御中兼具直接抗病毒作用与免疫调节功能，一方面抑制病毒复制，另一方面通过促进免疫细胞在炎症部位聚集以及激活免疫应答等方式增强宿主抗病毒免疫力。β-防御素还能作为疫苗佐剂提升免疫效果，在防治流感方面具有广阔的应用前景。
血凝素1(Hemagglutinins 1，HA1)：甲型流感病毒的血凝素1是病毒表面糖蛋白血凝素的一个关键亚型，主要负责病毒与宿主细胞的初始接触和吸附。血凝素1能特异性地识别并结合宿主细胞表面的唾液酸受体，触发病毒包膜与细胞膜融合，使病毒核衣壳进入细胞内并开始增殖。血凝素1具有高度可变性，其抗原位点易发生突变，导致抗原漂移或抗原转换，使得病毒能够逃避宿主体内已建立的特异性免疫反应，促进新病毒株的产生和传播。这种特性是流感病毒频繁引起季节性流行和大流行的重要原因之一。

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。
https://orcid.org/0009-0002-6390-5483(雷琪)

中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程

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

Abstract: 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 mouse β-defensin and interferon-γ when acting in mouse tracheal epithelial cells.
OBJECTIVE: To investigate the effect of recombinant hemagglutinin 1 on the production levels of mouse β-defensin and interferon-γ in 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 mouse β-defensin 2, 3, and 4, and interferon-γ were detected by qRT-PCR. The protein levels of mouse β-defensin 2, 3, and 4 were measured by enzyme-linked immunosorbent assay, and the protein expression of interferon-γ was 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 mouse β-defensin 2, 3, and 4 (P < 0.05) and interferon-γ in 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 mouse β-defensin 2, 3, and 4 and interferon-γ in mouse tracheal epithelial cells.

Key words: influenza A virus, recombinant hemagglutinin 1, β-defensin, interferon-γ, tracheal epithelial cells, mouse

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