| 流感嗜血杆菌下气道定植对哮喘小鼠肺组织TH17细胞表达及气道重塑的影响 |
| 投稿时间：2016-03-22 修订日期：2016-03-23 点此下载全文 |
| 引用本文：徐欣欣,董杨阳,杨玲,周妍,田烨,宋珍.流感嗜血杆菌下气道定植对哮喘小鼠肺组织TH17细胞表达及气道重塑的影响[J].医学研究杂志,2016,45(10):24-30,47 |
| DOI：
10.11969/j.issn.1673-548X.2016.10.007 |
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| 基金项目:国家自然科学基金资助项目（81200017） |
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| 中文摘要:目的 检测下呼吸道流感嗜血杆菌定植对哮喘小鼠TH17细胞的表达、气道炎症及对气道重塑的影响。方法 用卵清蛋白（ovalbumin，OVA）致敏和激发制备慢性哮喘小鼠模型，同时制作流感嗜血杆菌琼脂糖菌珠，在气道注菌之后的第1、7、21天处理小鼠，观察肺组织病理变化，流式细胞仪检测肺组织TH17细胞表达、测定CD4+T细胞TLR4的表达，并进行气道细菌培养和肺泡灌洗液（bronchoalveolar lavage fluid，BALF）细胞计数。结果 与正常组和哮喘非注菌组相比，哮喘注菌组有明显的哮喘症状和感染症状；病理学结果显示哮喘组气道黏膜水肿，管壁增厚，大量的炎性细胞浸润，注菌后炎症加重。哮喘小鼠气道注菌之后，随着时间的延长，气管壁厚度（WAt/Pbm）和平滑肌厚度（WAm/Pbm）均显著高于哮喘非注菌组。细菌学培养结果显示正常组第1、7天气管流感嗜血杆菌培养阳性，第21天阴性；哮喘组注菌后1、7、21天气管流感嗜血杆菌检测均阳性。流式细胞仪检测小鼠肺组织TLR4和TH17在CD4+T细胞的表达，哮喘组的TLR4和TH17表达均显著高于正常组，注菌之后二者表达显著增加，观察指标在哮喘注菌组变化更明显，差异有统计学意义（P<0.05）。BALF细胞计数结果显示，哮喘组BALF中细胞数明显高于正常组，注菌后细胞数进一步增多。相关性分析发现，哮喘组小鼠肺组织中CD4+T细胞TLR4的表达与BALF细胞总数呈正相关（r=0.746，P<0.05），TH17表达与CD4+T细胞TLR4的表达、气道WAt/Pbm、WAm/Pbm和BALF细胞总数呈正相关（分别r=0.612、0.611、0.537、0.752，P<0.05）。结论 流感嗜血杆菌下呼吸道定植可以通过激活TLR4，增加哮喘小鼠肺组织TH17细胞表达，加重哮喘气道炎症和气道重塑，可能是难治性哮喘预后的重要机制之一。 |
| 中文关键词:流感嗜血杆菌 哮喘 TLR4 TH17 气道重塑 |
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| Effect of TH17 Expression and Airway Remodeling in an Asthmatic Mice Model with Airway Colonizated by Haemophilus Influenza |
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| Abstract:Objective To detect TH17 expression, airway inflammation and airway remodeling in an asthma mice model with airway colonizated by haemophilus influenza, and discuss its potential signaling pathway. Methods Mice were sensitized and challenged by antigen ovalbumin(OVA)to built chronic mice asthma model, agar-beads enclosed haemophilus influenzae agarose were prepared at meantime. And then they were sacrificed on day1, day7 and day 21 after intratracheal instillation.Alterations of lung histopathology was observed. The percentages of TH17 and CD4+TLR4+ cells in total CD4+T cells from the lung were determined by flow cytometry. Airway bacterial culture, total cells counting ofbronchial alveolar lavage fluid (BALF) were performed. Results Compared with normal and non-incubated asthma groups, incubated asthma groups showed obvious asthma symptom and infection symptom. Airway thicking, mucosa edema, mucus secretion and inflammatory cells infiltration in lung tissue were obvious in asthmatic group, especially when after incubation. The ratios of total bronchial wall area to bronchial basement membrane perimeter (WAt/Pbm) and bronchial smooth muscle area to bronchial basement membrane perimeter (WAm/Pbm) significantly increased in the asthmatic group after incubation compared with normal and non-incubation asthma groups. Results of lung bacterial culture on day1, day7 and day21 after intratracheal inoculation were positive in asthma mice, and positive on day1 and day7, negative on day 21 in normal groups. Flow cytometry detected expression of TLR4 and TH17 on CD4+T cells revealed more expression in asthmatic groups than that in normal groups, and both significantly increased after incubation, especially in asthmatic groups. There was statistical difference between the normal groups and asthmatic groups. The numbr of total cells in BALF in asthma groups was significantly higher than that in normal groups, and more higher after incubation. Pearson correlation analysis revealed that the expression of TLR4 on CD4+T cells was significantly positively correlated with total cell numbers in BALF (r=0.746, P<0.05). The expression of TH17 was significantly positively correlated with expression of TLR4 on CD4+T cells, WAt/Pbm, WAm/Pbm and total cell numbers in BALF (r=0.612, 0.611, 0.537, 0.752, P<0.05). Conclusion The study showed that haemophilus influenza colonization in airway of asthmatic mice can elevate the expression of TLR4 on CD4+T cells, induce more TH17 cells in lung tissues, and accelerate airway inflammation and airway remodeling in asthma. This may be one of the mechanism to explain poor prognosis of refractory severe asthma. |
| keywords:Haemophilus influenza Asthma TLR4 TH17 Airway remolding |
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