【作者】 鲁士友；

【导师】 孙广仁；

【作者基本信息】 山东中医药大学 ， 中医基础理论， 2013， 博士

【摘要】 目的:根据前期的研究基础，对慢性阻塞性肺疾病(chronic obstructivepulmonary disease，COPD)及寒饮蕴肺证理论进行简要的梳理和论述，以肺组织中水通道蛋白AQP1及黏蛋白Muc5ac为切入点，进一步探讨COPD寒饮蕴肺证病机的动态演变与水通道蛋白AQP1及黏蛋白Muc5ac的关系，以丰富寒饮蕴肺证理论。本题延续采用寒凉刺激结合烟熏及气管内滴注内毒素（LPS）的方法建立COPD寒饮蕴肺证的病证结合大鼠模型，通过观察模型大鼠60天的自然演变过程及自拟温阳化饮方、布地奈德干预下的病机演变过程，阐释COPD寒饮蕴肺证模型大鼠肺组织水通道蛋白AQP1、黏蛋白Muc5ac、肺功能、肺组织形态学的动态变化（以10天为一个阶段），以及自拟温阳化饮方对COPD寒饮蕴肺证模型大鼠肺组织水通道蛋白AQP1、黏蛋白Muc5ac的调节作用，确定自拟温阳化饮方的作用靶点及时效，为该病证的防治提供新思路和理论支撑，指导预防措施的制定以及治疗时机选择和疗程的确定。方法:采用烟熏及气管内滴注LPS，配以寒凉刺激（0℃冰箱中冷冻，冰水混合物饮用）复制出COPD寒饮蕴肺证病证结合模型，动态观察（间隔10天，共60天）该大鼠模型的自然状态下的病机演变，并用自拟温阳化饮方和布地奈德对该病证模型进行动态干预，观察模型大鼠的一般情况，肺组织病理学，检测有关肺功能指标，肺组织中水通道蛋白AQP1，黏蛋白Muc5ac。结果:本题以前期规范性的程序复制出可靠的COPD寒饮蕴肺证大鼠模型，模型大鼠的形态及行为的表现、指标的变化与COPD寒饮蕴肺证的临床表现相符。造模后大鼠吸气阻力和呼气阻力升高、肺顺应性下降，肺组织中水通道蛋白AQP1表达降低，黏蛋白Muc5ac表达升高，病理切片显示肺组织出现炎性损伤和肺纤维化改变。在撤除造模因素后60天内，随着病程的延长模型大鼠的症状及各项指标没有明显好转，且出现恶化，在40天左右寒饮蕴肺证的表现弱化，有转变为其他证型的趋势。自拟温阳化饮方干预的模型大鼠症状逐渐好转，肺功能改善，水通道蛋白AQP1表达升高，黏蛋白Muc5ac表达降低，病理切片示炎性损伤明显减轻，在40天左右达到峰值，接近正常大鼠水平。布地奈德干预下的模型大鼠症状也有好转，但水通道蛋白AQP1表达升高不明显，黏蛋白Muc5ac也无明显降低，病理切片示炎性损伤减轻。结论:寒凉刺激结合烟熏和气管内滴注LPS建立的COPD寒饮蕴肺证大鼠模型是符合临床实际的理想的病证结合动物模型，具有良好的可靠性和稳定性。痰饮的形成是COPD寒饮蕴肺证病机演变的关键，水通道蛋白AQP1表达降低，黏蛋白Muc5ac表达升高与痰饮的形成关系密切。寒饮蕴肺证可能是COPD的早期或急性期证型，自拟温阳化饮方能够上调肺组织中水通道蛋白AQP1的表达和下调黏蛋白Muc5ac的表达，消除痰饮，对COPD寒饮蕴肺证有肯定的疗效，并且对该病证的治疗存在时效性，水通道蛋白AQP1、黏蛋白Muc5ac是其作用靶点之一。更多还原

【Abstract】 Objective: According to the previous basic research on chronic obstructivepulmonary disease (chronic obstructive pulmonary disease, COPD) and accumulation ofcold fluid in the lung syndrome theory, with AQP1water channel protein in lung tissue andmucin Muc5ac as the starting point,to further explore the COPD accumulation of cold fluidin the lung disease machine dynamic evolution and water channel AQP1and mucinMuc5ac relationship, in order to enrich the theory of accumulation of cold fluid in the lung.It continues by cold stimulation combined with smoking and intratracheal instillation oflipopolysaccharide (LPS) rat model with the method of COPD accumulation of cold fluidin the lung syndrome, evolution of pathogenesis of budesonide by observing the model rats60days of natural evolution and the warming drink, intervention, explains COPD colddrink lung syndrome model of rat lung tissue water channel protein AQP1, and mucinMuc5ac and pulmonary function, the corresponding lung histomorphology changes (10days for a stage), as well as the warming drink fabric water channel protein AQP1, andmucin Muc5ac role in the regulation of COPD accumulation of cold fluid in the lung ratmodel of Lung Group, determine the target and aging with warming drink side, to providenew ideas and theoretical support for the prevention and treatment of diseases, todetermine the guiding preventive measures and treatment timing and duration.Methods: The model with the COPD syndrome of cold fluid retention in lungdisease copied out by smoking and intratracheal instillation of LPS accompanied by coldstimulation (0℃refrigerator freezer, ice and water mixture drinking),dynamicobserve(interval of10days,60days) the rat model of the natural state of evolution, and the disease model for dynamic intervention with warming drink and budesonide, observe thegeneral model of rat, pulmonary pathology, lung function testing related indicators, AQP1water channel protein in lung tissue, mucin Muc5ac.Results: This program use preliminary normative to copy out the COPDaccumulation of cold fluid in the lung of rats model. Index form and behavior in model ratsof change and COPD accumulation of cold fluid in the lung syndrome are reliable toclinical manifestations. After modeling the inspiratory resistance and expiratory resistancerats increased, decreased lung compliance, water channel protein decreased expression ofAQP1in lung tissue, Muc5ac expression increased, pathology showed lung tissue changesin inflammatory injury and fibrosis. On the60day after removal of molding factors, alongwith the prolonged duration of symptoms and indexes of the model rats had no obviousimprovement, and deteriorated in40days or so, the weakening of accumulation of coldfluid in the lung syndrome, is transformed into the other card type trend. Rat model ofsymptoms by warming drink party intervention gradually improved, improve lung function,water channel protein AQP1expression increased, Muc5ac expression decreased, thepathological sections showed inflammatory injury was relieved, reached the peak at40days, close to the normal level in rats. Rat model of budesonide intervention symptomsalso improved, but the water channel protein AQP1expression did not increase remarkably,mucin Muc5ac has no obvious fdecrease, the pathological sections showed decreasedinflammatory injury.Conclusion: Cold stimulation combined with smoking and intratracheal instillation ofLPS is the establishment of animal model combining with clinical practical ideals diseasesof COPD accumulation of cold fluid in the lung of rats model of has a good reliability andstability. This is critical for the formation of COPD accumulation of cold fluid in the lungsyndrome pathogenesis evolution, reduced expression of water channel protein AQP1,mucin Muc5ac expression increased with the close relationship between the formation ofphlegm. Accumulation of cold fluid in the lung syndrome might be COPD early or acutesyndromes, with warming drink to the expression of AQP1water channel protein in lungtissue up-regulation and down-regulation of the expression of mucin Muc5ac, eliminatingphlegm, has a definite effect on COPD accumulation of cold fluid in the lung syndrome,and treatment of the disease in aging water channel protein, AQP1, Muc5ac mucin is oneof its targets.更多还原