Study On The Efficacy And Immunological Regulation Mechanism Of Astragalus And Asthma

Asthma is a kind of immune allergic diseases for the characteristics of reversible airway obstruction, airway hyper-responsiveness and airway inflammation caused by variety of factors. Asthma is a common disease that issevere harm to human health over the world, which is considered as one of the four major chronic diseases by WHO.The etiology and pathological mechanism of asthma are very complex, there are many factors involved in the pathogenesis of asthma, the etiology and pathogenesis of the disease has not yet entirely clear. The study of the pathogenesis of asthma by the western medicine found that there are mainly several mechanisms, such as the immunological mechanism, inflammatory mechanism, mechanisms of airway remodeling and airway neural regulation mechanism. In addition, glucocorticoid resistance is also one of the main pathogenesis to the intractable asthma. Genetic factors also play an important role in the pathogenesis of asthma.At present, the treatment of bronchial asthma is mainly basied on anti-inflammatory and antispasmodic. In the acute attack of asthma, glucocorticoid, theophylline and β2agonists can relieve airway spasm symptoms and control the progress of the disease. In the remission stage of asthma, it can be treated by atomization inhalation for topical anti-inflammatory. However, there are many disadvantages of western medicine in the treatment of asthma, such as the price is relatively high, the side effects are relatively serious and the treatment cycle is long, so it is difficult to be accepted by the majority of patients with bronchial asthma. The traditional Chinese Medicine for the treatment of asthma through clinical practice for thousands of years, has the characteristics of rich experience, diversification of the means of treatment, no obvious side effect etc, with the nearly natural therapies and significant effect, it is gradually accepted by the majority of patients with asthma.Compared with other diseases in Department of internal medicine, external treatment plays an important role in the treatment of asthma. External treatment of asthma is originated from 《Zhang Shi Yi Tong》 in the Qing Dynasty. The main characteristic of this therapy is the combination of acupoint sticking and the topical drug, not only retain the effect of the drug, but also correct the viscera Yin and Yang partial hyper-function or partial failure through the conduction and adjustment of main and collateral channels.To improve the operation of Qi andblood in the main and collateral channels, produce treatment and good regulating effect on asthma pathological state.MJPC cataplasm is a new kind preparation of traditional Chinese medicine, which is composed of Ephedra, white mustard seed, Rhizoma Corydalis, bitter almond, ginger juice, according to the characteristics of asthma with qi stagnation and blood stasis, In order to play a role in the treatment of asthma in the acute attack of asthma and in the remission stage of asthma. The study mainly focuses on the effect of MJPC cataplasm to the treatment of asthma on BN animal model and the regulation of Th1/Th2 and Th17/Treg imbalance, analyzing the treatment on asthma of MJPC cataplasm and its immune regulation mechanism.Objective1. Establish the BN rat asthma animal model, treat with MJPC cataplasm, in order to study the effect of MJPC cataplasm on the asthma animal model.2. Study the expression of Thl, Th2, Th17 and Treg cells related cytokines and proteins of the animal model, in order to explore the immunological regulation mechanism of MJPC cataplasm on asthma animal model.Methods:40 SPF BN rats fed for 3 days, randomly divided into four groups, blank control group, model control group, positive control group, MJPC cataplasm treatment group. On the fourth day, the four groups except the blank control group, intraperitoneal injection by 10% sensitizedOVA suspension for the first time. On the eleventh day, the four groups except the blank control group, intraperitoneal injection by 10% sensitizedOVA suspension for the second time. From the sixteenth day, positive control groupintraperitoneal injection of 1%dexamethasone (0.001ml/g); MJPC cataplasmtreatment group stick MJPC cataplasm on the GeShu and FeiShu. From the nineteenth day, the four groups atomization excitation with 1% OVA for seven days.After the atomization excitation, four groups BN rats detected lung function index under anesthesia.Then collect the bronchoalveolar lavage fluid(BALF), blood, spleen and lung tissue, for white blood cell counting and classification, Elisa detection of Thl, Th2, Th17 and Treg related cytokines, and flow cell counting and classification.Results:1. Pulmonary function test results:Compared to the model control group, the value of airway resistance in MJPC cataplasm treatment group is significantly lower(**P<0.01) than that in the model group when the concentration of Ach is 3.125mg/ml and 6.25mg/ml, when the concentration of Ach is 12.5mg/ml, the value of airway resistance in MJPC cataplasm treatment group is lower (*P<0.05) than that in the model group, the differences have statistical significance.2.The classification of WBC in whole blood count results:Compared to the model control group,the MJPC cataplasm treatment group of eosinophil and neutrophil number weresignificantly lower than that of model group(**P<0.01),2the differences have statistical significance.3. The classification of WBC in BALF count results:Compared to the model control group, the MJPC cataplasm treatment group of eosinophil and neutrophil were significantly lower than that of model group(**P<0.01), the differences have statistical significance. 4. The results of H&E staining of lung tissue sections:The model control group is visible in lung tissue inflammatory cell infiltration, Bronchial epithelial cells arranged in disorder, fall off, alveolar septal thickening. On the contrary, the MJPC cataplasm treatment group and the positive control group with a small amount of inflammatory cell infiltration, there was no obvious lung pathological injury. In the blank control group, there was no inflammatory cell infiltration, and the alveolar and bronchial epithelial structure is complete and clear.5. Flow cytometry results:Compared to the model control group, the number of Thl cells in MJPC cataplasm treatment group is higher than that of model group(*P<0.05), the number of Th2 cells in MJPC cataplasm treatment group is significantly lower than that of model group(**P<0.01), the number of Th17 cells in MJPC cataplasm treatment group is lower than that of model group(*P<0.05), the number of Treg cells in MJPC cataplasm treatment group is significantly higher than that of model group(**P<0.01).6. The results of immunohistochemistry:Compared to the model control group,the integral optical density of IFN-y in MJPC cataplasm treatment group was significantly higher than that in the model group(**P<0.01); the integral optical density of IL-4 in MJPC cataplasm treatment group was lower than that in the model group(*P<0.05); the integral optical density of IL-17 in MJPC cataplasm treatment group was significantly lower than that in the model group(**P<0.01);IL-10 integral optical density value and model group had no significant difference. For the immunohistochemistry of the four proteins pre control protein detection, compared to the model control group,the integral opticaldensity of T-bet in MJPC cataplasm treatment group was higher than that in the model group(*P<0.05); the integral optical density of GATA-3 in MJPC cataplasm treatment group was significantly lower than that in the model group(**P<0.01); the integral optical density of RORrt in MJPC cataplasm treatment group was significantly lower than that in the model group(**P<0.01); the integral optical density of FoxP3 in MJPC cataplasm treatment group was higher than that in the model group(*P<0.05).7. Related cytokines in serum, BALF and lung homogenate in Elisa testing results: compared to the model control group, there was no significant difference in the concentration of IL-4 in serum of MJPC cataplasm treatment group, theconcentrationof IL-4 in BALF is lower than that of model group(*P<0.05), the concentration of IL-4 in lung homogenate is significantly lower than those in the model group(**P<0.01). There was no significant difference in the concentration of IFN-γ in serum of MJPC cataplasm treatment group, the concentration of IFN-γ in BALF is higher than that of model group(*P<0.05), the concentration of IFN-γ in lung homogenate is significantly higher than those in the model group(**P<0.01). The concentration of IL-17 in serumis significantly lower than those in the model group(**P<0.01), the concentration of IL-17 in BALF is lower than that of model group(*P<0.05), the concentration of IL-17 in lung homogenateis significantly lower than those in the model group(**P<0.01). The concentration of IL-10 in serumis significantly higher than those in the model group(**P<0.01), the concentration of IL-10 in BALFishigher than those in the model group(*P<0.05), there was no significant difference in the concentration of IL-10 in lung homogenate between two groups.Conclusion:1. MJPC cataplasm has therapeutic effect on BN rat model of asthma.Pulmonary function detection showed that it can ease airway smooth muscle spasm induced by Ach.WBC classification and counting showed that it can significantly reduce the number of eosinophils and neutrophils and other inflammatory cells.H&E staining showed that it can reduce the injury of lung tissue in asthmatic animal.2. Immune regulation for BN rat asthma of MJPC cataplasm was by improving the imbalance of Th1/Th2 andThl7/Treg implementation. The results of flow cytometry, Immunohistochemistry and the results of related cytokines in serum, BALF and lung homogenate in Elisa testing showed that, there were low expression of Th2 and Th17, andhigh expression of Thl and Treg in MJPCcataplasm treatment group, while there were high expression of Th2 and Th17, and low expression of Thl and Treg in model control group.To sum up, MJPC cataplasm has therapeutic effect on BN rat model of asthma, and immune regulation for BN rat asthma of MJPC cataplasm was by improving the imbalance of Thl/Th2 and Thl7/Treg implementation. And the role of MJPC cataplasmin the treatment of asthma remains to be further confirmed in clinical.