ISSN 1674-3865  CN 21-1569/R
主管:国家卫生健康委员会
主办:中国医师协会
   辽宁省基础医学研究所
   辽宁中医药大学附属医院

中国中西医结合儿科学 ›› 2019, Vol. 11 ›› Issue (5): 382-386.doi: 10.3969/j.issn.1674-3865.2019.05.005

• 实验论著 • 上一篇    下一篇

雾霾环境下流感病毒感染小鼠致炎模型系统评价

敏娜,吴振起,刘光华   

  1. 110847 沈阳,辽宁中医药大学2016级中医儿科学专业研究生(敏娜);110032 沈阳,辽宁中医药大学附属医院儿科(吴振起);110847 沈阳,辽宁中医药大学基础医学院温病教研室(刘光华)
  • 出版日期:2019-11-01 发布日期:2019-11-08
  • 通讯作者: 吴振起,E-mail:zhenqiwu@163.com
  • 作者简介:敏娜(1992-),女,辽宁中医药大学2016级硕士研究生在读。研究方向:中医药防治感染性疾病
  • 基金资助:
    国家自然科学基金青年项目(81403293);辽宁省自然科学基金项目(2013020180)

Systematic evaluation of inflammatory model of mice infected by influenza virus in haze environment

MIN Na, WU Zhenqi, LIU Guanghua   

  1. Graduate School of Liaoning University of Traditional Chinese Medicine,Shenyang 110847,China
  • Online:2019-11-01 Published:2019-11-08

摘要:
目的
建立雾霾环境下流感病毒(IV)感染小鼠模型,探讨其对小鼠血清中肿瘤坏死因子α(TNF-α)、鼻咽灌洗液中气道分泌型免疫球蛋白A(sIgA)及肺组织中肺表面活性物质相关蛋白A(SP-A)表达的影响。
方法
将72只SPF级BABL/c小鼠随机分成正常组、PM2.5感染组、IV感染组、联合感染组,每组18只。除正常组外,对其余各组小鼠进行PM2.5气管滴注/IV感染模型处理,于造模后第3、7、10天进行取材。镜下观察小鼠肺组织病理切片;采用酶联免疫吸附试验(ELISA)测定各组小鼠血清中TNF-α、鼻咽灌洗液中sIgA水平变化;并运用免疫组织化学SP法及Western-blot法检测小鼠肺组织中SP-A蛋白的表达。
结果
经PM2.5及IV双重感染小鼠后,镜下可见肺泡间隔增厚,细支气管壁增厚,大量炎症细胞浸润;小鼠血清中TNF-α表达水平显著上调;小鼠鼻咽灌洗液中sIgA含量降低,肺组织中SP-A表达降低。与PM2.5感染组比较,IV感染组能够降低鼻咽灌洗液中sIgA及肺中SP-A的表达,各时间点比较差异无统计学意义(P>0.05);联合感染组在第3、7天能够显著降低sIgA、SP-A的表达,差异有统计学意义(P<0.05),并于第10天开始逐步升高。
结论
本实验中,3种造模方法均造成肺部不同程度的损伤,尤以PM2.5气管滴注联合IV感染所造成的肺部损伤最为严重。故证明此法简单、可靠,有利于进一步研究肺部疾病及致炎机制。

关键词: 雾霾, PM2.5, 造模, 流感病毒, 免疫损伤

Abstract: Objective
To establish a mouse model of influenza virus(IV) infection in haze environment, and to investigate its effect on the expression of tumor necrosis factor-α(TNF-α) in serum, airway secretory immunoglobulin A(sIgA) in nasopharyngeal lavage fluid and pulmonary surfactant-associated protein A(SP-A) in lung tissue of mice.
Methods
Seventy-two SPF grade BABL/c mice were randomly divided into the normal group (group A),PM2.5 tracheal drip group (group B),IV infection group(group C) and -PM2.5- tracheal drip+IV infection group(group D), with 18 rats in each group. Except the group A, the other groups of mice were treated with PM2.5 tracheal drip/IV infection model, and samples were obtained on the day of 3, 7, 10 after the model was established. The lung tissue sections were examined by histopathology; the serum levels of TNF-α and the nasopharyngeal lavage fluid sIgA in the mice were assessed by enzyme-linked immunosorbent assay(ELISA);the expression of SP-A protein in lung tissue of mice was detected by immunohistochemical SP method and Western-blot method.
Results
After double infection with PM2.5 and IV in mice, the pathological examination revealed thickening of alveolar septum and bronchioles, and extensive inflammatory cell infiltration in the lung tissues; the expression of TNF-α in serum of mice was significantly up-regulated; the content of sIgA in nasopharyngeal lavage fluid and the expression of SP-A in lungs of mice decreased. Compared with group B, group C could decrease the expression of sIgA in nasopharyngeal lavage fluid and SP-A in lung, and there was no significant difference at each time point(P>0.05);the expression of sIgA and SP-A was significantly decreased in group D on day 3 and 7(P<0.05),and it began to rise gradually on the tenth day.
Conclusion
In this experiment, all three methods of modeling have caused lung injury in varying degrees; especially, the lung injury caused by PM2.5 tracheal drip and IV infection is the most serious. Therefore, it is proved that this method is simple and reliable, which is helpful for further study of pulmonary diseases and inflammation mechanism.

Key words: Haze, PM2.5, Modeling, Influenza virus, Immune injury