Analysis Of The Composition Of An Chuan-fang In The Treatment Of Bronchial Asthma And The Influence On Th2/Treg Cell Subsets

Background:Bronchial asthma is one of the most common chronic respiratory diseases which is a chronic airway inflammation in involved variety of cells and cell components.Asthma is characterized by variable symptoms of wheeze,shortness of breath,chest tightness and/or cough,and by variable expiratory airflow limitation.Both symptoms and airflow limitation characteristically vary over time and in intensity.These variations are often triggered by factors such as exercise,allergen or irritant exposure,change in weather,or viral respiratory infections.China's incidence rate arrives 1%-5%and still rise.Especially in the past 20 years,asthma's morbidity and mortality increased significantly.The etiology mechanism of bronchial asthma is complex and the Th2/Treg response imbalance plays a very important role.It may provide new ideas and strategies for the treatment of bronchial asthma in the future.By detecting the changes of Th2/Treg cell subsets level,people can confirm the presence of airway inflammation and severity to assess the therapeutic effect of drugs.In traditional Chinese medicine asthma was belonged to "Xiao-Disease" category.The motherland medicine research on asthma has a long history which has been related to asthma-related symptoms,etiology and pathogenesis,treatment and other description.Chinese medicine point out that asthma has pods.That is,phlegm in the lungs,the case of exogenous,diet,emotions,fatigue,exercise and other factors,lead to phlegm obstruction and lung loss of"Qi" for asthma.Since ancient times,Chinese medicine has unique advantages on treatment of bronchial asthma.An-Chuan-Fang which is derived from many years of clinical practice and experience of Chinese medicine has a good effect in the treatment of colds type asthma.Granted a national patent in 2015,it has been made a preliminary study and admitted as hospital preparations.Objective:Two aspects of the study on clinical efficacy and animal experiments would be conducted on the basis of previous research.The aim of these studies was to confirm the therapeutic effect of An-Chuan-Fang on colds type bronchial asthma and to explore the material basis and possible molecular mechanisms of its treatment.These studies provide a theoretical basis for the clinical treatment effect of An-Chuan-Fang.Method:High performance liquid chromatography(HPLC)has been used to quantitatly analyze of chemical composition and the main components of An-Chuan-Fang.The study of the effect of Th2/Treg cell subsets in the mice with bronchial asthma.50 mice were divided into 5 groups.Normal group mice were fed with SPF forage.Bronchial asthma model mice were treated with physiological saline.Low-dose group bronchial asthma mice were treated with An-Chuan-Fang Chinese herbal medicine(12.5 g/kg)by intragastric administration.High dose group bronchial asthma mice were treated with An-Chuan-Fang Chinese herbal medicine(50.1g/kg)by intragastric administration.Positive control group bronchial asthma mice were treated with prednisone(7.5mg/kg).The amount change of Th2/Treg cell subsets on the expression of IL-4,IL-13,IL-10 and TGF-? protein in the serum of mice were studied by enzyme-linked immunosorbent assay(ELISA).Th2 and Treg cell subsets were detected by flow cytometry.100 cases of bronchial asthma at the onset of cold type asthma were divided into two groups according to the random numbers.The treatment group of patients was taked An-Chuan-Fang 1 dose/day combining with budesonide/formoterol powder inhalant 160/4.5ug,1 suction 2 times/day while the control group of patients only budesonide/formoterol powder inhalant.After four weeks' treatment,two groups were observed and compared by asthma control test(ACT)score,chest tightness,wheezing,lung wheezing,cough,sputum,and pulmonary functionResult:1.HPLC can quantitatly analyze the chemical composition of Anchuan Fang,including ephedra,ganoderma lucidum and licorice.The results showed that there was a good linear relationship between ephedrine hydrochloride at 40.9ug/ml and 409.0ug/ml,while the pseudoephedrine hydrochloride showed a good linear relationship between 20.6ug/ml and 206.0ug/ml.The HPLC chromatograms of ephedrine hydrochloride and pseudoephedrine hydrochloride were precisely drawn.2.After treatment with An-chuan-fang,compared with the normal control group,the administration group model animal's lung tissue pathological sections showed that lung tissue and tracheal wall lesions were light.The proportion of Th2 cells in the blood of the model group was significantly lower than that of the normal control group(P<0.01)and the dose was correlated(P<0.01),that was to say,the higher the dose of An-chuan-fang,the lower the proportion.In contrast,the proportion of Treg cells in the blood of the mice treated with the control group was significantly higher than that of the control group(P<0.01)and the dose was correlated(P<0.01).Compared with the control group,the levels of IL-4 and IL-13 in the serum of asthmatic mice were obviously decreased with significant differences(P<0.01)and dose-dependent effect(P<0.01).While compared with the control group,the levels of anti inflammatory factor IL-10 in the serum of asthmatic mice increased significantly(P<0.01)in the administration group and had a dose dependent effect(P<0.01).There was no significant difference in the levels of TGF-? between the asthma group and the control group(P>0.05).3.Asthma control rate comparison.After 4 weeks of treatment,the complete control rate of two groups with mild asthma was better than the same group(P<0.05).And the control rate of patients with moderate asthma was better than that of the control group(P<0.05).Comparison of therapeutic effects of traditional Chinese medicine.The control rate of the two groups was significantly higher than that of the control group(P<0.05).After 2 weeks the odds ratio of the two groups was obviously higher than that of the control group(P<0.05)and the difference was significant(P<0.05).This difference was further expanded after four weeks(P<0.01).After 4 weeks of treatment,the remission rate of chest tightness in the treatment group of mild asthma patients was significantly higher than in the control group(P<0.05).After 2 weeks of treatment,the respite control rate of both two groups with moderate asthma was better than the same group and the difference was further expanded after 4 weeks(P<0.01).The control rate of patients with mild asthma was better than that of the moderate group after 4 weeks(P<0.05).After treatment,the wheezing remission rate of both two groups with moderate asthma was better than than the same group(P<0.05).After 2 weeks of treatment,the wheezing remission rate in the treatment group of mild asthma patients was much higher than in the control group(P<0.05)and the same phenomenon occured to the moderate asthma patients after 4 weeks(P<0.05).There was no significant difference in the rate of cough and sputum between the two groups(P<0.01).The improvement rate of pulmonary function in the two groups was significantly higher than that in the same group(P<0.01),but the treatment group was no better than the control group whether mild or moderate asthma(P>0.05).Adverse reactions.There were 4 cases:1 case of tremor in the treatment group,2 cases of tremor in the control group and 1 case of pharynx stimulation in the control group.The adverse reaction rate of the two groups had no difference(P>0.05).Conclusion:1.The chemical composition and the main components of An-Chuan-Fang treatment of bronchial asthma may be mainly ephedrine hydrochloride and pseudoephedrine hydrochloride.2.That An-Chuan-Fang can be used to therapy bronchial asthma may be reduce of Th2 cell subsets of the differentiation level and improve the relative content of Treg cells and restore the asthma state Th2/Treg cell subgroup imbalance.The mechanism also lies on inhibiting the inflammation associated with asthma factors such as IL-4,IL-13 and promoting the factor IL-10 secretion which inhibit asthma.So the airway inflammation may be reduced and the airway hyperresponsiveness may be relieved.3.Compared with the control group,An-Chuan-Fang combined with western medicine budesonide/formoterol in the treatment of bronchial asthma has a significant advantage.An-Chuan-Fang was confirmed the efficacy by the clinical research.lt is the effective prescription on treatment of cold typel asthma.