摘要
目的：慢性阻塞性肺疾病(chronic obstructive pulmonary disease, COPD)是一种常见的慢性呼吸系统疾病,以气道、肺实质和肺血管的慢性炎症为特征,可以导致气流受限不完全可逆。该病程呈进行性发展,与肺部对香烟烟雾等有害气体或颗粒的异常炎症反应有关。气道的慢性炎症持续存在导致气道壁损伤和修复过程反复发生。目前全世界COPD死亡率居所有死因的第四位,其致死率呈逐年上升的趋势。对患者个体来说,临床表现除肺部症状外,常常伴有全身系统的严重疾病表现,严重影响着患者的劳动能力和生活质量。由于本病患病人数多,死亡率高,社会经济负担较重,COPD现已成为一个重要的公共卫生问题。
中医药治疗COPD具有丰富的临床经验,尤其是针对COPD稳定期的治疗,本着“治未病”的中医理论思想,从预防复发角度出发,采用扶正固本类中药,不仅可以减轻患者的临床症状、提高机体免疫力,还可以减少急性发作次数,从而延缓COPD的病程进展速度。
固本止咳中药,从COPD本虚标实的病机特点入手,采用补益肺、脾、肾三脏之气,兼顾痰热、血瘀的特点达到扶正以祛邪的功效。前期临床研究显示,采用固本止咳中药观察30例患者取得了较好的临床疗效。其中改善患者咳嗽、咳痰的总效率达100%,改善喘息有效率达85.71%,综合疗效评定总有效率96.67%,在改善患者肺功能、调节机体免疫等方面有较好效果。探讨固本止咳中药治疗COPD的作用机理,研究固本止咳中药对呼吸道黏膜免疫系统的影响机制,阐述呼吸道黏膜免疫首道防线作用与中医正气“卫外”功能的关系,提出中医“治未病”理论在COPD防治应用中的优越性。
方法：本实验采用鼻腔滴入脂多糖(LPS)加熏香烟的方法建立COPD小鼠动物模型,实验分为空白对照组、模型对照组、固本止咳中药组；运用HE染色、电镜对形态学进行观察,评价COPD模型小鼠的病理改变,及固本止咳中药对COPD模型小鼠病理改变的影响；使用动物肺功能仪分析系统检测实验小鼠的呼吸生理变化,分析固本止咳中药对COPD模型小鼠肺功能的影响；应用ELISA方法检测小鼠气道slgA分泌水平,应用免疫组织化学方法检测实验小鼠肺组织中NE含量表达情况,探讨固本止咳中药对COPD模型小鼠气道及肺组织炎症状态的影响。
结果：1.采用鼻腔滴入LPS加熏香烟的方法建立的COPD小鼠模型,可使小鼠气道管腔变窄,管腔内分泌物增加,支气管黏膜上皮脱落,气管微绒毛倒伏,粘缩成团；肺组织间质内炎性细胞浸润增加,肺泡壁结构破坏、溶解破裂,呈囊状扩张,肺泡孔增大,肺毛细血管床明显减少,形成肺大泡,多为生理无效腔等均符合COPD病理学改变。经固本止咳中药治疗后小鼠气道黏膜及肺组织炎性浸润有所改善,肺毛细血管床相对丰富,但仍存在肺大泡改变；微结构变化显示,气道黏膜结构改善明显,肺泡上皮细胞结构仍存在病理改变。2.反应小鼠气道阻塞程度的FEVO.1、PEVO.2,及小气道功能状态的FEF25-75、PEF等指标显示,模型对照组较空白对照组降低(P<0.05),固本止咳中药组较模型对照组增高(P<0.05)。3.BALF中sIgA的含量检测可反应小鼠气道炎症状态,结果显示模型对照组较空白对照组sIgA明显升高(P<0.05)；固本止咳中药组较模型对照组sIgA分泌量明显降低(P<0.05)；固本止咳中药组较空白对照组略有升高(P>0.05)。4.NE作为中性粒细胞活化后释放的一种毒性分子,可破坏肺组织弹性纤维,促使形成肺大泡。空白对照组显示NE分泌含量极少,模型对照组小鼠NE阳性表达显著增多,其阳性面积显著高于空白对照组与固本止咳中药组(P<0.01),空白对照组与固本止咳中药组比较无统计学差异(P>0.05)。
结论：采用鼻腔滴入LPS加熏香烟的方法可成功复制COPD动物模型。固本止咳中药可明显改善COPD模型小鼠的气流受限程度,其中以改善小气道功能更为明显；可使呼吸道sIgA分泌含量明显降低,使COPD模型小鼠气道慢性炎症得到有效控制,减轻了sIgA的过度免疫应答,从而起到调节呼吸道黏膜局部免疫功能的保护作用；同时使肺组织中NE含量显著减少,肺部炎性浸润得到有效控制。其中sIgA分泌含量与NE含量表达呈正相关性。固本止咳中药通过减轻COPD模型小鼠的气道黏膜及肺组织炎症浸润程度,从而调整呼吸道黏膜免疫状态,对局部气道炎症的控制可减轻气道壁的损伤和修复过程的发生,起到改善小鼠气道通气功能障碍的作用。我们推测,固本止咳中药通过抑制COPD模型小鼠的慢性炎症状态,降低气道黏膜的过度免疫应答,促进气道黏膜的损伤修复,使黏膜免疫达到平衡状态,起到类似中医“正气”卫外的防御功能作用。从而达到控制疾病进程,预防急性发作的目的,体现中医“治未病”理论“既病防变,瘥后防复”的思想。
Objective:Chronic Obstructive Pulmonary Disease (COPD) remains a major public health problem, is characterized by persistent airflow limitation, which is usually progressive and associated with an enhanced chronic inflammatory response in the airways and the lung to noxious particles or gases. The chronic inflammation in the airways causes structural changes resulting from repeated injury and repair. It is the fourth leading cause of death in the world. The patients suffer from not only the the respiratory symptom but the constitutional symptom, COPD is a major cause of chronic morbidity and mortality in the world and results in a social and economic burden, which is both substantial and increasing.
Traditional Chinese Medicine has a wealth of clinical experience in the treatment of COPD, Especially in the stable stage of this disease. By the methord of strengthening healthy and improving immunity with in the theory of "preventive treatment", which could relieve the clinical symptoms of patients, improve the immunity, decrease acute exacerbations and slow down the couse of the disease.
Gubenzhike recipe has the function of invigorating the lung and kidney, strengthening the spleen and reducing phlegm, and dispel evils. Early clinical studies showed Gubenzhike recipe has good clinical efficacy. The total efficiency improvement in patients with cough, sputum was100%, improve asthmatic effective rate was85.71%, the total efficiency of comprehensive evaluation was96.67%. To observe the effect and mechanism of Gubenzhike recipe how to enhance the respiratory passage mucosa immune system, and discuss the common materials between respiratory passage mucosa immune system and the theory of wei-energy.
Methods:The mouse COPD model was established by exposure to cigarette smoke in combination with intra-nostrils injection of lipopolysaccharide. The mice were divided into three groups:negative control group, model group and Gubenzhike recipe group; Use the methods of HE eosin staining and electronic microscope to observe the morphology characteristic of COPD model; Test the pulmonary function by animal pulmonary function machine; Use the ELISA to test the slgA level in respiratory tract; Use the method of Immunohistochemistry to test the expression of NE in lung tissue.
Results:1.The mouse COPD model was established by exposure to cigarette smoke in combination with intra-nostrils injection of lipopolysaccharide. The morphology characteristic is up to COPD, the airway narrowed, intraluminal secretions increased, bronchial epithelial cilia exfoliated, tracheal microvilli lodged and sticky huddled. Inflammatory cell infiltrate in the Pulmonary interstitial tissue, structure of alveolar destructed and even dissolved and ruptured, cystic di latation, alveolar pores distructed, pulmonary capillary bed significantly reduced, and shaped the bullae. Gubenzhike recipe has the partly function of repair the pathological changes.
2. Gubenzhike recipe has the function of partily repair the lung tissue morphology; The FEV0.1, FEV0.2, FEF25-75, PEF and the content of sIgA in the upper respiratory tract which in Gubenzhike recipe group had significant difference compared with those in model group (P<0.05).
3. The expression of NE in lung tissue is decreased in Gubenzhike recipe group (P<0.01). The negative control group had no significant difference compared with Gubenzhike recipe group (P>0.05).
Conclusion:The mouse COPD model was established by exposure to cigarette smoke in combination with intra-nostrils injection of lipopolysaccharide. Gubenzhike recipe might improve the pulmonary function especially the small airways, and respiratory tract mucosa immunity of COPD model. The inflammation of the airways can be controlled, reduced the immune response. In this way, it can regulate the function of repiratory passage mucosa immune system and reduce the inflammation of the lung tissue. There was a positive correlation between sIgA and NE. Gubenzhike recipe alleviates the inflammatory infiltration state of the COPD mouse models' airway mucosa and lung tissue, therefore adjusts respiratory mucosal immune status. Contained local airway inflammation can reduce the airway wall damage and repair process which is the main reason of the mouse models' imoroved airway dysfunction. We assume that the Gubenzhike recipe inhibits COPD mouse models' chronic inflammatory state, as well as airway mucosal immune response, which process resembles the defense function of the healthy energy (zheng qi). Furthermore it can slow down the evolution of diseases prevention acute exacerbations, which reflets the idea" To prevent, progression once ill, to prevent relapse once cured" in the traditional Chinese medicine theory.
引文
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