| BackgroundChronic obstructive pulmonary disease (COPD) is a common chronic respiratory disease with high rate of disability and mortality, with a characteristic of Incomplete airflow limitation. As reported, this disease has ranked to the fourth cause of death, and is expected to be the world’s fifth economic burden of disease in2020. Some pathogenic factors, such as infections, smoking, air pollution, occupational dust harmful gas and so on, can lead to chronic inflammation of airway, lung parenchyma, and pulmonary vasculature. But many kinds of inflammatory cells,such as, neutrophils, alveolar macrophages, lymphocytes,can release various bioactive substances(e.g. IL-1、IL-4、IL-8、TNF-α) to promote the occurrence and development of chronic airway inflammation. The airway mucus hypersecretion and inflammatory reaction is considered to be one of the important causes of COPD, which eventually can lead to airway obstruction and airflow limitation.COPD belongs to lung disease category in Chinese traditional medicine of cough, asthma", phlegm, whose root is deficiency of air. Zhang Zhongjing in Treatise pointed out that typhoid table puzzled, heart gas, retching, fever and cough, or thirsty, or profit, or choking, or dysuria, less abdominal fullness, or asthma, which all can be treated by Xiaoqinglong Decoction. Golden Eagle and treatment of phlegm and cough pointed that Xiaoqinglong Decoction can treat the disease of anasarca, when the person is sweating. Xiaoinglong Decoction is an important prescription for treating drinking outside in cold syndrome. The composition of Xiaoqinglong Decoction is Ephedra sinica, cinnamon twig, Herba Asari, Rhizoma Pinelliae, Rhizoma Zingiberis, Fructus Schisandrae Chinensis, radices paeoniae alba, Radix liquiritiae. Ephedra sinica can deal with sweating, asthma, and diuresis, and cinnamon twig can improve the force of dispersion. The combination of cinnamon twig and peony can recuperate the body. This method is often used for the treatment of diseases of the respiratory system. Professor Chen Baotian continues the ancient experience, and has achieved good effect on the clinical treatment of COPD with sanxiaotang. Sanxiaotang includes Koshiba Koyu, Xiao Xianxiong Decoction and Xiaoqinglong decoction, which is the commonly used formula in the treatment with the cold drink. Professor chen thought that Sanxiaotang can be widely used to treat the COPD. Our research group plan to establish animal model of COPD, and explore the effect of sanxiaotang on chronic obstructive pulmonary disease in the aspects of preventing cough, phlegm, asthma, allergy. On the basis of pharmacodynamics, we further study the related pharmacological mechanism of sanxiaotang in the treatment of COPD. Interleukin-4(IL-4) secreted by II T helper cell (Th2cell) has many kinds of Biological effects, such as, the activation of B cells and T cells proliferation, differentiation of CD4+T cells into II T helper cell and so on. IL-4can participate in COPD pathogenesis by regulating the eosinophilic inflammation, promoting airway fibrosis, inducing airway mucus hypersecretion and airway hyperresponsiveness (AHR). Interleukin-8(IL-8) is the main group of chemotactic factor’s family, is important mediator of many inflammatory diseases, has close relationship with the occurrence and development of COPD, and is a better indicator to monitor and assess the therapeutic effect. TNF-a is a proinflammatory cytokine with widely biological activity of. Smoking and other risk factors of COPD can lead to bronchial epithelial cell injury, increase TNF-a secretion, promote the inflammatory cell chemotaxis and activation, leading to airway structure damage. Studies suggest that many factors lead to the increase of serum level of TNF-a in COPD.Aquaporins waterchannel protein(AQPs) is a membrane channel protein family, responsible for water rapid transmembrane transportation in various tissues and cells. AQP1is mainly expressed in airway peripheral vascular, lymphatic and alveolar capillary endothelial cells and visceral pleural mesothelial cells. In this study, we detect the expression of AQP1in lung in order to explore the effect of sanxiaotang on pulmonary edema.We tried to explore the possible effect mechanism of sanxiaotang on COPD through three important indexes:interleukin-4, interleukin-8, tumor necrosis factor. At the same time, we observed cough and breathing frequency, pipe diameter and observation of pulmonary pathology images in the COPD rat in order to explore the effect of sanxiaotang on cough, asthma, allergy. The aim of the research is to provide the scientific basis for sanxiaotang development. Degree of improvement of clinical symptoms and safety evaluation were also observed in the patients with chronic obstructive pulmonary disease.Part One The construction of COPD rat model and intervention effect of sanxiaotangObjectiveExplore the pharmacodynamics of sanxiaotang on COPD. To construct COPD model in Wistar rats; to observe the influence of sanxiaotang with three different doses on cough and respiratory frequency, the percentage of nuclear cells in bronchoalveolar lavage fluid, cytokines in serum, lung bronchiectasis and water channel protein expression in COPD rat.Methods1. Healthy Wistar rats of clean grade, adaptive feeding for7d, were randomly divided into normal group, model group, low dose group, middle dose group, high dose group and positive control group (ambroxol group),8rats in each group.2. Using injection of LPS and smoke in trachea method to construct the model of chronic obstructive pulmonary disease for4weeks. Each group were given different treatment, sanxiaotang high dose group (10g/kg), middle dose group (5g/kg), low dose group (2.5g/kg), ambroxol tablets control group (10.4mg/kg), model control group (saline) and normal control group (physiological saline), free diet.3. Observe cough and breathing frequency of the experimental animal per unit time in the process of drug treatment and1weeks after the treatment.4. Sacrifice the rats, extract the bronchoalveolar lavage fluid to detect various percentage of mononuclear cells, in order to further analysis of anti-allergic effect.5. Normal lung tissue section, check the bronchiectasis level.6. IL-4, IL-8, TNF-a ELISA kits are used to detect IL-4, IL-8, TNF-a of serum in rat.7. The two step immunohistochemical technique of Envision is used to detect water channel protein1(AQP1) expression in the lung tissue.8Statistic methods(1) Measurement results can be represented by mean±SD (2) different groups before and after treatment was compared by SPSS13statistical software package using the analysis and one-way ANOVA method, comparison between groups apply the LSD method; if the variance not neat, approximate F test Welch method will be applied, and Dunnett’s T3is used to comparison between groups. The rats cough frequency, respiratory frequency before and after treatment are compared using analysis of covariance; P<0.05for the difference was significant.Results1. One weeks after modeling, rats in each group showed the symptoms of cough, shortness of breath.2. Prior to the administration of three low dose group, middle dose group, high dose group, western medicine group, model group, cough frequency (F=0.220, P=0.925), respiratory rate (F=6.879, P=0.000) had no significant difference, but the difference compared with the normal group had statistical significance (P<0.05), suggested that symptoms in rats after building the model is significantly different from before. After administration, the cough frequency (F=49.040, P=0.000), respiratory rate (F=12.990, P=0.000) overall group were significant different. Sanxiaotang high dose group, middle dose group, low dose group, western medicine group compared with the model group all were statistically significant, suggesting sanxiaotang can improve the cough and shortness of breath in chronic obstructive pulmonary disease rats. 3. One week after treatment, lymphocyte percentage solution (F=15.091, P=0.000), percentage of neutrophil (F=11.534, P=0.000), percentage of eosinophils (F=3.045, P=0.020), basophil percentage (F=2.479, P=0.047) in rats bronchoalveolar were statistically significant among the groups, suggested that sanxiaotang can effectively reduce the increased lung cells percentage, reduce the allergic reaction.4. Lung bronchiectasis was statistically significant (F=18.557, P=0.000) among groups. Sanxiaotang low dose group, middle dose group, high dose group, western medicine group compared with the model group, the difference was statistically significant (P=0.000, P=0.000, P=0.000, P=0.000), suggested that the sanxiaotang can effectively alleviate intrapulmonary bronchial dilatation level in chronic obstructive pulmonary disease rat.5. After administration of the content of IL-4(F=15.078, P=0.000),the content of IL-8(F=29.442, P=0.000), the content of TNF-a(F=18.648, P=0.000) in serum of rats in each group were statistically significant among groups.Sanxiaotang low dose group, middle dose group, high dose group, western medicine group, normal group compared with the model group the differences were statistically significant, suggested that the sanxiaotang can effectively reduce the content of serum IL-4, IL-8, TNF-a and the inflammatory reaction.6. AQP1expression in the lung of the overall group is statistically significant (F=34.297, P=0.000). Sanxiaotang low dose group, middle dose group, high dose group, western medicine group and model group were statistically different, suggested that the sanxiaotang can effectively promote the water channel protein1(AQP1) expression in lung, thereby increase the removal and transport of lung water, improve water metabolism, reduce the state of pulmonary edema, and effectively protect acute lung injury.DiscussionTraditional Chinese medicine sanxiaotang can effectively improve clinical symptoms of the chronic obstructive pulmonary disease, reduce the proportion of lung cells, reduce the allergic reaction, effectively improve IL-4, IL-8, TNF-a content, reduce inflammation, can alleviate the situation of bronchiectasis pulmonary in lung tissue and promote the up-regulation of expression of aquaporins1, thereby increase the clearance and transport of lung water, improve water metabolism, reduce the state in order to reduce the loss of lung tissue of pulmonary edema.Part Two Clinical observation of sanxiaotangObjectiveObserve the effect of sanxiaotang on patients with chronic obstructive pulmonary disease in the aspects of improvement degree of clinical symptoms and safety evaluation.MethodsAll cases conform to the society of respiratory system formulation of the chronic obstructive pulmonary disease diagnosis and treatment guideline of Chinese Medical Association (2007Edition). Diagnostic criteria:patients from respiratory department of Internal Medicine Department of traditional Chinese medicine, Nanfang hospital outpatient and ward, sex not limited, at the age of48-70years old, a total of60cases, were randomly divided into control group and the observation group,30cases each group. The control group was treated with bed rest, antibiotics, bronchodilators, reduce airway resistance, glucocorticoid, phlegm and other conventional treatment of Western medicine. The observation group were treated with sanxiaotang on the basis of the treatment of the control group.Sanxiaotang:Asarum10g, Pinellia tuber10g, Licorice10g, Schisandra10g, dried ginger10g, Cassia twig10g, Ephedra10g, Radix Paeoniae Alba10g, Bupleurum20g, Scutellaria15g, Codonopsis10g, Ginger10g, Jujube10g, Coptis chinensis10g, Trichosanthes30g, Houttuynia cordata30g.The patients in the two groups were under three conventional tests, liver function, renal function tests before and after treatment. We assess the clinical symptom according to the guiding principle of research on new drugs.All data were analyzed with statistical software SPSS13.0.(1) Measurement results can be represented by mean±SD.(2)The treatment efficacy of the two groups was compared with two independent samples nonparametric tests. The comparison of cytokine content in the serum after treatment applied analysis of covariance. Each group before and after treatment was compared using analysis of variance (One-way ANOVA) for statistical analysis. When the variance is neat using LSD-t multiple comparison test; variance not neat, with approximate Welch method of analysis of variance, P<0.05was statistically significant.Results1Fourteen days after treatment, the total efficiency of the observation group and the control group was93.33%and73.33%respectively. The observation group was better than control group, there was significant difference between two groups (P=0.039). This suggested that on the basis of conventional treatment, sanxiaotang can more effectively improve the clinical symptoms of chronic obstructive pulmonary disease of the patients, improve the quality of life.2The comparison of IL-4, IL-8, TNF between routine western medicine treatment group and sangxiaotang+western medicine treatment group before treatment had no statistically significant different. IL-4(F=10.874, P=0.002), IL-8(F=378.696, P=0.000), TNF a (F=404.358, P=0.000) after treatment have statistical differences. This suggested that the sanxiaotang can improve the content of IL-4, IL-8, TNF in the serum of patients with chronic obstructive pulmonary disease, and can reduce the inflammatory reaction.DiscussionSanxiaotang combined with conventional western medicine in the treatment of chronic obstructive pulmonary disease has more curative effect than the simple routine treatment of Western medicine, while it’s safe and reliable. Therefore, sanxiaotang has important clinical significance and potential clinical value. |
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