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崔爽,张明倩,梁五林,郭凡帆,张天睿,欧文静,伍永鸿,贾占红,旦增曲培,张硕峰.LPS诱导慢性支气管炎急性发作大鼠模型的建立和评价[J].湖南中医药大学学报英文版,2022,42(11):1830-1836.[Click to copy
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| LPS诱导慢性支气管炎急性发作大鼠模型的建立和评价 |
| 崔爽,张明倩,梁五林,郭凡帆,张天睿,欧文静,伍永鸿,贾占红,旦增曲培,张硕峰 |
| (北京中医药大学中药学院, 北京 102488;西藏藏医药大学, 西藏 拉萨 850000) |
| 摘要: |
| 目的 通过多次气道内雾化给药的方式建立脂多糖（lipopolysaccharide, LPS）致大鼠慢性支气管炎急性发作（acute exacerbations of chronic bronchitis, AECB）模型，探讨LPS诱导慢性支气管炎急性发作的可行性。方法 将36只SD大鼠按体质量随机分为空白组、模型组、复方甘草组。采用多次气道内雾化吸入LPS的方法建立大鼠AECB动物模型。采用肺功能仪评估乙酰胆碱（Ach）和组胺激发大鼠的气道高反应，肺泡灌洗液（bronchoalveolar lavage fluid, BALF）采用细胞计数板进行白细胞总计数，瑞氏染色法进行白细胞分类计数（分为两类：中性粒细胞、巨噬细胞），HE、PAS、Masson染色观察肺组织的病理形态学变化，ELISA法检测BALF中白介素6 （interleukin 6, IL-6）和肿瘤坏死因子-α（tumor necrosis factor-α, TNF-α）含量。结果 与空白组相比，模型组大鼠注射不同剂量氯化乙酰胆碱和组胺（1∶1）混合液后气道阻力显著升高(P＜0.01)；BALF中白细胞总数、单核巨噬细胞和中性粒细胞数量明显增加(P＜0.01)；支气管黏膜上皮存在部分脱落坏死，管腔变窄且腔内出现较多炎性分泌物，伴肺泡塌陷，肺泡间隔增厚和肺出血，大量炎性细胞浸润，气道黏膜杯状细胞增生，出现大量胶原纤维沉积；BALF中促炎因子IL-6、TNF-α水平升高(P＜0.01)。复方甘草组较模型组可明显减轻AECB大鼠肺组织的炎症损伤和肺出血程度，减轻气道上皮杯状细胞增生以及气道壁周围胶原纤维沉积，改善肺功能等。结论 多次气道内雾化吸入LPS可成功建立大鼠AECB模型。 |
| 关键词: 慢性支气管炎 慢性支气管炎急性发作 动物模型 气道高反应 黏液高分泌 气道重塑 |
| DOI：10.3969/j.issn.1674-070X.2022.11.010 |
| Received:July 06, 2022 |
| 基金项目:西藏自治区科技计划项目（XZ201801-GA-16）。 |
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| Establishment and evaluation of rat model with LPS-induced acute exacerbation of chronic bronchitis |
| CUI Shuang,ZHANG Mingqian,LIANG Wulin,GUO Fanfan,ZHANG Tianrui,OU Wenjing,WU Yonghong,JIA Zhanhong,Danzeng Qupei,ZHANG Shuofeng |
| (School of Chinese Materia Medica of Beijing University of Chinese Medicine, Beijing 102488, China;Xizang University of Tibetan Medicine, Lasa, Xizang 850000, China) |
| Abstract: |
| Objective To establish a rat model of lipopolysaccharide (LPS)-induced acute exacerbation of chronic bronchitis (AECB) established by multiple intra-airway nebulization administration, and to explore the feasibility of LPS-induced acute exacerbation of chronic bronchitis. Methods A total of 36 SD rats were randomly divided into blank group, model group, and compound Gancao group according to their body mass. An animal model of AECB in rats was established by multiple intra-airway nebulized inhalation of LPS. The airway hyperresponsiveness of rats caused by acetylcholine (Ach) and histamine was assessed by pulmonary function instrument. Bronchoalveolar lavage fluid (BALF) was used for total white blood cell (WBC) count by cell counting plate. Wright's staining was used for WBC differential count (two categories: neutrophils, macrophages). HE, PAS and Masson staining were used to observe the pathomorphological changes of lung tissue, and ELISA was applied to detect the content of interleukin 6 (IL-6) and tumor necrosis factor-α (TNF-α) in BALF. Results Compared with blank group, the airway resistance was significantly higher in the model group after injection of different doses of acetylcholine chloride and histamine 1∶1 mixture (P＜0.01); the total number of WBC, mononuclear macrophages and neutrophils in BALF significantly increased (P＜0.01); there was partial exfoliation and necrosis in the bronchial mucosa epithelium, narrowing of the lumen and more inflammatory secretion in the lumen, accompanied by alveolar collapse, thickening of alveolar septum and pulmonary hemorrhage; there was a large number of inflammatory cells infiltration, airway mucosa goblet cells proliferation, and mass collagen fiber deposition; the levels of proinflammatory cytokines IL-6 and TNF-α in BALF increased (P＜0.01). Compared with model group, the compound Gancao group showed significant decrease in the inflammatory injury and the degree of pulmonary hemorrhage in the lung tissue of AECB rats, less proliferation of airway epithelial goblet cells and deposition of collagen fiber around the airway wall, and improved lung function, etc. Conclusion Multiple intra-airway nebulized inhalation of LPS can successfully establish a model of acute exacerbation of chronic bronchitis in rats. |
| Key words: chronic bronchitis acute exacerbation of chronic bronchitis animal model airway hyperresponsiveness mucus hypersecretion airway remodeling |
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