| [Objective]1. To looking for the acting targets of Yiqi Ditan Huayu recipe in regulating immune balance by experimental research on Yiqi Ditan Huayu recipe,that is the experience recipe of Yunnan province famous traditional Chinese medicine doctor in curing asthma.2. Establishment of asthmatic mouse model, to explore mechanism of Yiqi Ditan Huayu recipe in curing asthma by observing the effecets of the recipe regulate the balance of Thl/Th2and Th17/Treg.[Methods]1. Establishment of asthmatic mouse model:50SPF female BALB/C mice were randomized into the normal control group, asthma model group and Yiqi Ditan Huayu recipe high, medium and low dose groups respectively. The mice in the normal control group and drug intervention groups were intraperitoneally injected with sensitized solution (contains50u g OVA)on the1st,8th and15th day, and stimulating solution with2%OVA were consecutively given by aerosol inhalation from the22nd to28th day to stimulate the asthma attack. Sensitization and stimulation were performed with physiological saline in the normal control group.2.Drug intervention experiment:During the stimulating period (the22nd to28th day), Yiqi Ditan Huayu recipe were intragastrically given every day to the mice in the intervention groups30minutes before the aerosol inhalation of2%OVA. Physiological saline was intragastrically given in the normal control and asthma model group.3.Index detection:Behavioral changes of all the mice were observed. Lung tissue biopsies were made and examined under a microscopy. BALF was collected for white cell differential count. The serum of the mice was collected and concentrations of IFN-Y, IL-4, IL-4/IFN-γ and IL-17were determined in ELISA. [Results]1. Behavior ObservationThe symptoms and physical signs of the mice were alleviated by different extent in the3drug intervention groups compared with those in the normal control group. The symptoms and physical signs were significantly improved. During the feeding period, the fur was glory, the body weight increased significantly and only1mouse died, so the overall situation was relatively good.2. Pathological ChangesAccording to the lung tissue biopsy:compared with the asthma group, inflammatory cells around intrapulmonary bronchioles and blood vessels decreased in the3drug intervention groups, especially in the group treated with medium dose. The airway wall was complete and secretions from airway epithelial mucus decreased.3. Aggregation of airway inflammatory cellsCompared with the mice in the model group, the total white blood cell count as well as EOS, NEU and LYM differential count in Bronchoalveolar Lavage Fluid (BALF) decreased in the drug intervention group with medium dose; while in the drug intervention groups with high and low dose, the total WBC count as well as EOS, NEU and LYM differential count in BALF also showed decrease, but the differences were not statistically significant.4. Cytokine levels in serumCompared with the normal model group, in the drug intervention groups with high and medium does, serum IFN-γ level increased (P<0.05), IL-4level decreased (P<0.05), IL-4/IFN-γ ratio decreased (P<0.05), IL-17level decreased greatly (P<0.01). In the drug intervention groups with low does, serum IFN-γ level increased (P<0.05), IL-4level decreased (P>0.05), IL-4/IFN-γ ratio decreased (P<0.05), IL-17level decreased greatly (P<0.01), and the differences were statistically significant. The differences of IFN-γ level, IL-4level, IL-4/IFN-γ ratio and IL-17level within the intervention groups were not statistically significant (P>0.05) [Conclusions]1. Yiqi Ditan Huayu recipe could treat the bronchial asthma by reversing the Th1/Th2and Th17/Treg imbalance, the immune unbalance caused by Th2and Th7advantage.2. Yiqi Ditan Huayu recipe could alleviate acute asthmatic airway inflammation lesions to some extent by intervening the number of airway inflammatory cells and expression level of inflammatory cytokines, which preliminarily indicated that the mechanism of the prescription was reversing... |
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