Effects Of Guben Cough And Chinese Herbal Medicine On γδT Cells And Related Cytokines In Lung Tissue Of Mice With Chronic Obstructive Pulmonary Model

Objective:Chronic obstructive pulmonary disease (COPD) is a kind of common diseases,which is characterized by persistent airflow limitation. Sustained chronic inflammation make airway remodeling by repeated injury and repair,and it is the main reason of irreversible progress of this disease.The morbidity and mortality rate of COPD has increased year by year and seriously affect the patients quality of life, cause social and economic burdens. Traditional Chinese medicine (TMC) treatment of COPD has certain advantages, under the guidance of Chinese medicine theory "preventive treatment", by the Chinese medical herbs of strengthening healthy, TMC could relieve the clinical symptoms of patients and enhance the immune function, decrease times of exacerbations and slow down the progression of the disease.Gubenzhike recipe accord with the clinical characters and pathogenesis in TCM of COPD, has the efficacy of reinforcing the deficiency and strengthening the genuine. Our early clinical studies showed Gubenzhike recipe has good clinical effect on reducing cough and expectoration, animal experiment demonstrate that it can relieve airflow limitation, reduce slgA in COPD mice, in so the inflammation of the airways can be controlled, the excessive immune response can be reduced, In this way, it can improve the state of respiratory passage mucosa immune system(RMIS), suggested that pathological process of COPD is closely related to espiratory passage mucosa immune.As a key effective cell of RMIS, y8T cell can produce various cytokines like KGF IL-17、Thl type cytokine IFN-yand Th2 type cytokine IL-4 etc. after excited, then play a role in regulating inflammation and promote wound healing, immune surveillance and maintain homeostasis It is important on maintaining the integrity of the respiratory mucosa, improve respiratory mucosal immune function.So study on the effect of Gubenzhike recipe on the biological activity of γδT cell and discuss the relationship between γδT cell and wei-energy will be helpful to further discussion the mechanism of Gubenzhike recipe treatment of COPD, propose that enhance the respiratory mucosal immune barrier function is an effective way to achieve the the theory of "preventive treatment" in TCM.Methods The mice were divided into three groups:negative control group, model group and Gubenzhike recipe group;The mouse COPD model was established by exposure to cigarette smoke in combination with intra-nostrils injection of lipopolysaccharide. Use the methods of HE eosin staining to observe the morphology characteristic of COPD model; Use the methods of mmunohistochemistry to test the expression of KGF in lung tissue of COPD model;Use the method of Flow cytometry to test the expression of epithelialy γδT cell in lung. ELI SA was used to investigate the IL-17 level in supernatant of epithelial lymphocyte culture, KGF level in supernatant of lung homogenate, IFN-y and IL-4 level in serum.Results 1.The morphology characteristic of mice COPD model is consistent with clinical pathological changes of COPD.2. The proportion of γδT cells in lung epithelial T lymphocytes in each experimental group:The proportion of γδTT cells of model group is higher than negative control group(P< 0.05),and the proportion of γδT cells of Gubenzhike recipe group is lower (P<0.05); 3. The IL-17 level in supernatant of epithelial lymphocyte culture in each experimental group:it is higher in model group than negative control group(P < 0.05),and lower in Gubenzhike recipe group (P<0.05), it’s coincides with the changing trend of γδT cell.4.The expression of KGF in lung tissue in each experimental group:it is decreased in model group (P<0.05),and is higher in Gubenzhike recipe group than those in model group (P<0.05).5. IFN-y and IL-4 level in serum in each experimental group:The serum level of IFN-y decreased and IL-4 increased significantly in Gubenzhike recipe group than that in model group (P< 0.05)Conclusion Gubenzhike recipe can reduce the proportion of γδT cells in T cells,that in lung epithelial cells of COPD model mice. To reduce the excessive immune response of γδT cells and Improve the local respiratory mucosa of mice inflammatory condition. Gubenzhike recipe can influence of the following related cytokines by adjust γδT ymphocytes bioactivities:First of all, Gubenzhike recipe can reduce IL-17 content in the culture medium of lung lymphocyte of mice model of COPD, thereby reducing the airway inflammation, reducing the occurrence of inflammation in the airway wall damage process, improving mucus hypersecretion, delaying gas road reconstruction. At the same time, can promote the secretion of lung tissue in mice COPD model KGF. Upregulation of KGF expression in lung tissue. Promoting repair of inflammatory injury of lung tissue. Improved airway mucosal integrity. Enhanced mechanical barrier function of the respiratory tract mucosa immunity. In addition, Gubenzhike recipe can decrease the content of serum IFN-y,elevating the level of IL-4,correction of Th1/Th2 imbalance,alleviating airway inflammation in mice, slowing emphysema formation, to delay COPD disease processes.We speculate that Gubenzhike recipe can effectively enhance the defense and supervising functions of Wei Qi by upright support, adjust the γδT clls biological activity, and regulatory cytokines produced by it. control of airway mucosa local inflammation injury. Promote the repair process after injury, to improve the function of respiratory mucosa integrity, playing a role in maintaining tissue homeostasis. That is, to achieve the body "Yinping Yangmi" state, enhance the ability of the body to resist the evils, at the same time by reducing the damage repair repeatedly, delaying the formation of airway remodeling and pulmonary emphysema, so as to achieve the purpose of control process of COPD disease. It embodies the thought of TCM theory,that is "Preventive treatment of disease" of "which lead to disease recurrence after cure."...