Effect of chronic <italic style="font-style: italic">Aspergillus fumigatus</italic> exposure on the expression of Toll-like receptor 4 in the airway of rats with bronchial asthma

Zhang Lingxuan; Li Meng; Song Ge; Gao Fusheng

doi:10.13406/j.cnki.cyxb.003766

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Effect of chronic Aspergillus fumigatus exposure on the expression of Toll-like receptor 4 in the airway of rats with bronchial asthma

Basic research | 更新时间：2025-12-31

- Effect of chronic Aspergillus fumigatus exposure on the expression of Toll-like receptor 4 in the airway of rats with bronchial asthma
- Journal of Chongqing Medical University Vol. 50, Issue 12, Pages: 1702-1709(2025)
- 作者机构：
  
  1.山东第二医科大学临床医学院，潍坊 261000
  2.山东第二医科大学附属医院呼吸与危重症医学科，潍坊 261000
- 作者简介：
- 基金信息：
- DOI：10.13406/j.cnki.cyxb.003766
  CLC： R562.2+5
- Received：27 May 2024，
  
  Revised：2024-07-13，
  
  Accepted：02 September 2024，
  
  Published：28 December 2025
- 稿件说明：
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张令轩,李萌,宋歌等.慢性烟曲霉暴露对支气管哮喘大鼠气道Toll样受体4表达的影响[J].重庆医科大学学报,2025,50(12):1702-1709.

Zhang Lingxuan,Li Meng,Song Ge,et al.Effect of chronic Aspergillus fumigatus exposure on the expression of Toll-like receptor 4 in the airway of rats with bronchial asthma[J].Journal of Chongqing Medical University,2025,50(12):1702-1709.
张令轩,李萌,宋歌等.慢性烟曲霉暴露对支气管哮喘大鼠气道Toll样受体4表达的影响[J].重庆医科大学学报,2025,50(12):1702-1709. DOI： 10.13406/j.cnki.cyxb.003766.

Zhang Lingxuan,Li Meng,Song Ge,et al.Effect of chronic Aspergillus fumigatus exposure on the expression of Toll-like receptor 4 in the airway of rats with bronchial asthma[J].Journal of Chongqing Medical University,2025,50(12):1702-1709. DOI： 10.13406/j.cnki.cyxb.003766.

摘要

目的

探讨慢性烟曲霉暴露对支气管哮喘大鼠气道Toll样受体4（Toll-like receptor 4，TLR4）表达的影响。

方法

40只雄性Wistar大鼠随机分为5组（每组8只）。对照组（A组）：给予卵清蛋白（ovalbumin，OVA）致敏和生理盐水激发。支气管哮喘组（B组）：使用OVA诱导致敏和激发的方式构建大鼠哮喘模型。支气管哮喘+烟曲霉孢子吸入1周组（C组）、2周组（D组）和3周组（E组）：哮喘大鼠每周给予2次通过鼻腔吸入烟曲霉孢子的处理，分别吸入1周、2周和3周。测定血清总IgE水平及肺泡灌洗液（bronchoalveolar lavage fluid，BALF）中炎症细胞总数及分类计数、白细胞介素-5（interleukin-5，IL-5）、白细胞介素-13（interleukin-13，IL-13）、γ-干扰素（interferon-γ，IFN-γ）和转化生长因子β（transforming growth factor-β，TGF-β）水平；肺组织切片行苏木素-伊红（hematoxylin and eosin，HE）、过碘酸雪夫（periodic acid schiff，PAS）和Masson三色染色，并行病理图像形态学测定和分析；免疫组化、逆转录聚合酶链式反应（reverse transcription-PCR，RT-PCR）及蛋白质免疫印迹（Western blot，WB）检测大鼠气道TLR4的表达。

结果

支气管哮喘大鼠吸入烟曲霉孢子3周后血清总免疫球蛋白E（immunoglobulin E，IgE）［（109.76±18.11） ng/mL］、BALF中炎症细胞总数［（5.88±1.04）×10

］及淋巴细胞［（2.27±0.37）×10

］、嗜酸性粒细胞计数［（1.49±0.43）×10

］、［IL-5（28.79±6.67） ng/mL］、IL-13［（118.63±20.51） ng/mL］及TGF-β［（105.00±15.73） ng/mL］、气道炎性细胞数（76.11±9.10）、上皮细胞损伤脱落百分比［（18.78±3.11）%］、杯状细胞增生程度［（19.98±2.42）%］及上皮下纤维化程度［（30.44±5.56）%］较吸入前差异有统计学意义（

0.05）。各组大鼠的TLR4/β-actin值与气道上皮损伤脱落程度呈正相关（

=0.960，

0.001），与气道杯状细胞增生程度呈正相关（

=0.944，

0.001），与气道上皮下纤维化程度呈正相关（

=0.856，

0.001）。

结论

慢性烟曲霉暴露可剂量依赖性上调支气管哮喘大鼠气道TLR4表达，进而加剧气道炎症和气道重构。

Abstract

Objective

To investigate the effect of chronic

Aspergillus fumigatus

（AF） exposure on the expression of Toll-like receptor 4 （TLR4） in the airway of rats with bronchial asthma.

Methods

A total of 40 male Wistar rats were randomly divided into control group （group A），bronchial asthma group（group B），bronchial asthma+1-week AF spore inhalation group（group C），bronchial asthma+2-week AF spore inhalation group（group D），and bronchial asthma+3-week AF spore inhalation group （group E），with 8 rats in each group. The rats in group A were sensitized with ovalbumin （OVA） and challenged with normal saline，and those in group B were sensitized and challenged with OVA to establish a rat model of asthma； the rats in groups C，D，and E were given nasal inhalation of AF spores twice a week for 1，2，and 3 weeks，respectively. The serum level of total IgE was measured，as well as the total and differential counts of inflammatory cells and the levels of interleukin-5（IL-5），interleukin-13（IL-13），interferon-γ（IFN-γ），and transforming growth factor-β（TGF-β） in bronchoalveolar lavage fluid（BALF）； lung tissue sections were used for HE，PAS，and Masson staining，and pathomorphological measurement and analysis were performed； immunohistochemistry，RT-PCR，and Western blot were used to measure the expression of TLR4 in the airway of rats.

Results

After the inhalation of AF spores for 3 months in rats with bronchial asthma，there were significant increases in the serum level of total IgE［（109.76±18.11） ng/mL］，the total number of inflammatory cells［（5.88±1.04）×10

］ and the counts of lymphocytes［（2.27±0.37）×10

］ and eosinophils［（1.49±0.43）×10

］ in BALF，the levels of IL-5［（28.79±6.67） ng/mL］，IL-13［（118.63±20.51） ng/mL］，and TGF-β［（105.00±15.73） ng/mL］ in BALF，the number of inflammatory cells in the airway （76.11±9.10），the percentage of epithelial cell detachment［（18.78±3.11）%］，the degree of goblet cell hyperplasia［（19.98±2.42）%］，and the degree of subepithelial fibrosis［（30.44±5.56）%］（

0.05）. The TLR4/β-actin value was positively correlated with the degree of epithelial cell detachment（

=0.960，

0.001），goblet cell hyperplasia（

=0.944，

0.001），and subepithelial fibrosis（

=0.856，

0.001） in the airway of rats with asthma.

Conclusion

Chronic AF exposure can upregulate the expression of TLR4 in the airway of rats with bronchial asthma in a dose-dependent manner，thereby aggravating airway inflammation and airway remodeling.

关键词

Keywords

references

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Department of Pain，Xi’an Central Hospital

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Department of Respiratory Disease，Children’s Hospital of Chongqing Medical University；National Clinical Research Center for Child Health and Disorders； Ministry of Education Key Laboratory of Child Development and Disorders；China International Science and Technology Cooperation Base of Child Development and Critical Disorders；Key Laboratory of Children’s Vital Organ Development and Diseases of Chongqing Municipal Health Commission

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Effect of chronic Aspergillus fumigatus exposure on the expression of Toll-like receptor 4 in the airway of rats with bronchial asthma

DOI：10.13406/j.cnki.cyxb.003766

摘要

Abstract

关键词

Keywords

references