慢性阻塞性肺疾病大鼠模型建立与氨茶碱干预的研究

发布时间：2018-03-16 22:21

本文选题：慢性阻塞性肺疾病　切入点：氨茶碱　出处：《浙江大学》2010年硕士论文　论文类型：学位论文

【摘要】： 背景: 慢性阻塞性肺疾病(chronic obstructive pulmonary disease,COPD)是常见的呼吸系统慢性疾病,是一种以呼吸道吸入有害气体或有害颗粒致不完全可逆气流受限为特征并呈进行性发展的肺部非特异性炎症性疾病,是世界慢性病防治重点之一。COPD是当今危害人民健康主要疾病之一,发病率日益上升,由于病程迁延,且反复发作急性加重,严重影响了COPD患者的生活质量,目前已成为各国医疗主要负担之一。但是其具体发病机制仍不清楚,现在有包括气道和肺部炎症,蛋白酶/抗蛋白酶失衡,氧化/抗氧化失衡等学说。茶碱属黄嘌呤类衍生物,为经典的支气管扩张药物,其作用主要是通过抑制腺苷酸磷酸二脂酶(PDE)而实现,近年研究茶碱具有抑制炎症反应和免疫调节等作用。本实验采用烟熏及气管内滴入脂多糖建立COPD大鼠模型,并以氨茶碱大、小剂量对其干预,观察相关结果及分析有关机制。研究目的: 本实验利用烟熏及气管滴入脂多糖建立COPD大鼠模型。观察COPD大鼠在肺功能、肺泡灌洗液、大鼠血白细胞计数与分类以及肺部组织的病理改变,同时观察灌入氨茶碱大鼠在上述方面变化,研究氨茶碱在对COPD大鼠中的治疗作用。研究方法: 利用熏香烟及气管滴入脂多糖法建立COPD大鼠模型(COPD组),二级雄性Sprague-Dawleg(SD)大鼠(清洁级),随机分为对照组、模型组、氨茶碱小剂量组、氨茶碱大剂量组共四组,在不同时间点分批处死,进行大鼠肺功能、肺泡灌洗液(BALF)、血中炎细胞计数和分类,并观察肺组织的病理形态学改变。结果: 1.动物造模情况。实验动物在实验过程中的表现:模型组及各治疗组大鼠早期出现躁动不安并且先后出现咳嗽、气急、精神萎靡、行动迟缓、痰多纳少、毛发枯黄等症状,之后出现体重减轻,进食进水量逐渐减少等表现。模型组大鼠肺体积略增大,正常对照组则始终未见上述表现。 2.COPD模型组气道阻力明显高于对照组,而其肺顺应性却较对照组明显下降;肺泡扩大融合、肺泡数明显减少;气道阻力和肺顺应性之间有显著的负相关性。 3.病理形态学观察光镜观察健康对照组大鼠支气管肺组织上皮结构完整,支气管纤毛排列整齐,肺泡结构完整,在纤毛结构间夹杂有少数杯状细胞,管壁内有少数散在的淋巴细胞。COPD模型组肺组织支气管纤毛柱状上皮细胞部分剥脱,纤毛粘连倒伏变性和坏死,有的纤毛明显脱落,杯状细胞增生,黏膜下层及肌层大量炎性细胞浸润,可见支气管平滑肌明显增厚,管腔内充满大量中性粒细胞、肺泡巨噬细胞及黏液分泌物,肺泡扩张融合,肺泡隔明显增厚,其中可见炎细胞浸润和毛细血管扩张充血。 4.COPD模型组BALF内的白细胞总数、淋巴细胞数、中性粒细胞数均明显较对照组增多,但巨噬细胞增多不明显。中性粒细胞数量与白细胞总数呈显著正相关(r=0.873,P＜0.01)中性粒细胞数量与淋巴细胞数量亦呈显著的正相关性(r=0.712,P＜0.01)。 5.模型组大鼠与对照组大鼠血白细胞总数、巨噬细胞数、淋巴细胞数差别有显著性意义见表3.3。结合表3.2发现BALF中性粒细胞数量与血中性粒细胞呈显著正相关(r=0.675,P＜0.01)。 6.氨茶碱大、小剂量组均显示了较好的改善COPD大鼠模型肺功能的作用:气道阻力下降,肺顺应性提高;均有一定的抗炎作用,对气道重塑形态学的改变亦有一定的改善作用:抑制了大鼠模型各级气道管壁的厚度。 7.大剂量氨茶碱组较小剂量组对COPD大鼠的肺功能、抗炎作用、病理形态学的改善作用更明显。结论: 本实验所建立的COPD大鼠模型符合人类COPD的病理形态学特点。慢性气道炎症的反复刺激引起气管黏膜下腺体增生肥大,淋巴细胞、浆细胞等炎症细胞浸润,支气管纤毛柱状上皮层增厚,皱褶高耸,杯状细胞增生,支气管腔内中性粒细胞及黏液蓄积,平滑肌明显增厚,黏膜下及外膜胶原纤维增生,终末呼吸细支气管炎症明显,腔内常有中性粒细胞聚集管壁纤维性增生增厚及平滑肌层增厚,即气道重塑,最终导致气流受限。氨茶碱对缓解COPD大鼠气道壁增厚、减轻炎症细胞浸润,改善肺功能等起到重要作用。其中大剂量组在上述方面的作用更明显。
[Abstract]:Background:
Chronic obstructive pulmonary disease (chronic obstructive pulmonary disease, COPD) is a common chronic respiratory diseases, is a kind of inhalation of harmful gases or particles caused by incompletely reversible airflow limitation is characterized by a progressive lung nonspecific inflammatory disease, is the world's chronic disease prevention and control is one of the key of.COPD the main diseases harm people's health, the incidence rate is rising, due to the prolonged course and recurrent episodes of acute exacerbation, seriously affecting the quality of life of patients with COPD, has become one of the main countries bear negative medical care. But its specific pathogenesis is still unclear, now include airway and lung inflammation, protease / antiprotease imbalance, etc. oxidant / antioxidant imbalance theory. Theophylline is a xanthine derivative, as a bronchodilator classic, its main function is through inhibition of adenylate phosphate Two lipase (PDE) has been achieved. In recent years, theophylline has inhibitory effects on inflammation and immune regulation. In this study, COPD rats were induced by smoking and intratracheal instillation of lipopolysaccharide, and the intervention was carried out with aminophylline at large or low doses, and the related results were observed and the mechanism was analyzed.
The purpose of the study is:
Smoked and intratracheal instillation of lipopolysaccharide in the rat model of COPD was constructed by using this experiment. COPD rats was observed in lung function, bronchoalveolar lavage fluid, the change of rat white blood cell count and classification of lung tissue pathology, and observe the changes of rats injected into aminophylline in the above aspects, the study of ammonia alkali in tea treatment effect on COPD in the rat.
Research methods:
The use of cigarette smoking and intratracheal instillation of lipopolysaccharide in the rat model of COPD was established (COPD group), two (SD) male Sprague-Dawleg rats (clean grade), were randomly divided into control group, model group, low-dose aminophylline group, aminophylline high-dose group four group, were killed in batches at different time point, lung the function of rat alveolar lavage fluid (BALF), inflammatory cell count and classification in the blood, and to observe the pathological changes in lung tissue.
Result:
1.. The animal model of experimental animal during the experiment showed: model group and treatment group rats appeared to be restless and has a cough, shortness of breath, listlessness, slow, sputum doner less, brown hair and other symptoms, after weight loss, decrease of food intake and inflow model. The lung volume of rats increased slightly, the normal control group has no such performance.
The airway resistance of 2.COPD model group was significantly higher than that of the control group, while the lung compliance was significantly lower than that of the control group. The alveolar enlargement and fusion and alveolar number decreased significantly, and there was a significant negative correlation between airway resistance and lung compliance.
3. pathological observation under light microscope, healthy control group lung tissue of rats with bronchial epithelial structural integrity, cilia arranged neatly, alveolar structure integrity, cilia structure with few goblet cell wall, there were a few scattered lymphocytes in.COPD model group lung tissue of bronchial ciliated columnar epithelial cells partially exfoliation, adhesion lodging degeneration and necrosis of cilia, some obvious cilia shedding, goblet cell hyperplasia, submucosa and muscular layer infiltration of inflammatory cells, visible bronchial smooth muscle thickening, the lumen filled with a large number of polymorphonuclear cells, alveolar macrophages and mucus secretion, alveolar expansion fusion, marked thickening of alveolar septa, which shows infiltration of inflammatory cells and telangiectasia congestion.
The total number of white blood cells in 4.COPD model group in BALF lymphocyte number, neutrophil counts were significantly increased compared with the control group, but the increase is not obvious. Macrophages were positively related to the total number of neutrophils and white blood cells (r=0.873, P < 0.01) number of neutrophils and the number of lymph cells also showed a significant positive correlation (r=0.712, P < 0.01).
5., there was a significant difference in the total number of white blood cells, macrophages and lymphocyte counts between the model group and the control group. According to table 3.3. combined with table 3.2, the number of BALF neutrophils was positively correlated with the blood neutrophils (r=0.675, P < 0.01).
6. a large group of small dose aminophylline, showed improvement in COPD rat model of pulmonary function better effect: decreased airway resistance, pulmonary compliance improved; anti-inflammatory effects of morphological changes on airway remodeling also have certain effect: the rat model of airway wall thickness at all levels decreased.
The small dose group of 7. large doses of aminophylline group had more obvious effect on the improvement of lung function, anti inflammatory and pathomorphology of COPD rats.
Conclusion:
The COPD rat model established in this experiment with the pathological features of human COPD. Repeated stimulation of chronic airway inflammation caused by tracheal submucosal gland hypertrophy, lymphocytes, plasma cells and other inflammatory cell infiltration, thickening of bronchial ciliated columnar epithelium, a tall, goblet cell hyperplasia, intraluminal neutrophils and mucus the accumulation of smooth muscle thickening, mucosa and adventitial collagen fiber hyperplasia, terminal respiratory bronchiolitis patients, cavity often neutrophil aggregation fibrous wall and hyperplasia of smooth muscle layer thickening, namely the airway remodeling, resulting in airflow limitation. Aminophylline to alleviate COPD rat airway wall thickening, inflammatory cell infiltration, improve play an important role in pulmonary function. The effect of high dose group in the above aspects of the more obvious.

【学位授予单位】：浙江大学
【学位级别】：硕士
【学位授予年份】：2010
【分类号】：R563.9;R-332

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