Clarify The Mechanism About How To Treat The Disease Of CARAS By Yi Qi Qu Feng Xuan Bi Hua Yin Formula On Regulating The Signal Transduction Pathway Of TLR9/AP-1

1 Object:Yiqi Qufeng, Xuan Bi Hua Yin Formula by content of AP-1 in the intervention TLR9/AP-1 signaling pathway and downstream leukocyte interleukin factor to charify the mechanism.of treatment of CARAS.2 Method:The guinea pig CARAS models which are established by OVA+Al(OH)3 intraperitoneal injection and OVA atomization inhalation are randomly divided into 4 drug intervention groups, model group, and a blank control group, a total of 6 groups, the Yiqi Qufeng, Xuanbi Formula is divided into high dose group, middle dose group and low dose group, western medicine group was intervented by dexamethasone. Guinea pigs are executed after 2 weeks intervention, left the guinea pig serum, lung tissue and nasal mucosa tissue and bronchoalveolar lavage fluid, 1.lung tissues and nasal mucosa tissues are observed by HE staining; 2. Giemsa staining is used to observe the lung tissue and bronchoalveolar lavage fluid eosinophilic granulocyte infiltration; 3, Western blotting is used to detect lung tissue homogenate of c-jun, c-fos, IL-4, IL-5, IL-6 expression; 4, PCR is used to detect c-jun and c-fos gene expression; 5, ELISA method is used to detect serum’s IL-4, IL-5 and IL-13.3 Results:3.1 HE stainingNasal mucosa:model group shows epithelial cell denaturation destroys, cilia necrosis, submucosal inflammatory cell infiltration; QFXBF high dose group, middle dose group, low dose group and dexamethasone group can been seen nasal ciliated epithelial cells and different degree of destruction and loss. Blank control group’s epithelial cells and cilia are good, no obvious inflammatory cell infiltration was found.Lung tissue:model group show bronchial alveolar destruction, bronchial wall thickening, and accompanied by exudation, bronchial submucosa and muscular layer, lung interstitial matter can be seen a large number of eosinophils and lymphocyte infiltration of inflammatory cells; QFXBF high dose group, middle dose group, low dose group and dexamethasone group, the lung tissue visible varying degrees of bronchial wall thickening and effusion, bronchial submucosa and muscular layer, lung interstitial can been seen different levels of eosinophils eosinophils, lymphocytes and other inflammatory cells infiltration.3.2 Giemsa stainingLung tissue and bronchoalveolar lavage fluid (BALF):model group shows a large number of eosinophils eosinophil infiltration. QFXBF high dose group, middle dose group, low dose group and dexamethasone group’s lung tissue can be seen different degrees of addicted to eosinophil infiltration. The model group showed a large number of eosinophils in the high dose group, middle dose group, low dose group and dexamethasone group. There is no obvious abnormality in the lung tissue and bronchoalveolar lavage in the blank group.3.3 ELISA detection of IL-4, IL-5, IL-13IL-4:four treatment groups and model group are significantly different (P<0.01);between low dose of QFXBF group and dexamethasone group have difference (P<0.05), and among the QFXBF three groups show no differences(P> 0.05); QFXBF middle dose group compares high dose group and dexamethasone group with no significant difference (P> 0.05). IL-5:four treatment groups are significantly different from model group (P< 0.01);between low dose of QFXBF group and dexamethasone group have difference (P< 0.05), and among the QFXBF three groups show no difference (P>0.05); between QFXBF middle dose group and high dose group, dexamethasone group show no significant difference (P>0.05). IL-13:four groups treatment group and model group are significantly different (P<0.01), low dose of QFXBF group compared the other three groups with difference in the treatment groups, and the three QFXBF groups compar with the presence of difference (P< 0.05), comparing QFXBF lowgroup with the high dose of QFXBF group,dexamethasone group shows the presence of significant difference (P < 0.01);middle of QFXBF group and high dose of QFXBF group compare no significant difference (P>0.05), but comparing with the dexamethasone group showed the presence of significant difference (P<0.01); high dose of QFXBF group and dexamethasone group have no difference.3.4 Western Blotting tests C-Jun, C-Fos, IL-4, IL-5 and IL-13In C-Jun and C-Fos group, drug intervention groups are significant difference in the model group (P< 0.01); Dexamethasone group and high, medium and low QFXBF groups are no significant difference. In IL-4 group, QFXBF low dose group compares with model group,and there are no significant difference (P>0.05); Model group and high dose group, high dose group and dexamethasone group have significant difference (P<0.01);between middle dose and high dose group of QFXBF groups, dexamethasone group, there are no significant difference (P>0.05). In IL-5 group, there is no significant difference between high dose group and dexamethasone group (P>0.05); Middle dose and low dose group have significant difference (P<0.01); Between QFXBF middle dose group and low dose group,there are no obvious difference (P>0.05), compared with the high dose group,there had significant difference (P<0.01). In IL-13 groups, model group and drug treatment of four groups have significant difference (P<0.01); QFXBF middle dose and low dose group show difference (P<0.05);comparing QFXBF low dose group with high dose group and dexamethasone group,there have significant difference (P<0.01); QFXBF middle dose group,high dose group and dexamethasone group have no significant difference (P>0.05).3.5 PCR detects the expression of C-Jun and C-Fos genesIn the C-Fos group, QFXBF groups,dexamethasone group and model group have significant difference (P< 0.01). There have different between QFXBF low dose group and QFXBF high dose group,so is dexamethasone group (P<0.05); QFXBF middle dose group, high dose group and dexamethasone group have no significant difference. In C-Jun group, there are significant differences between the QFXBF groups,the dexamethasone group and the model group (P< 0.01). Low dose of QFXBF group compared with QFXBF high dose group and dexamethasone group, there exist significant difference (P< 0.01); and compared with QFXBF middle dose group, there have no significant difference; QFXBF middle dose group compared high dose group with no significant difference, QFXBF middle dose group compares dexamethasone group with significant difference (P<0.01);QFXBF high dose group and dexamethasone group have no significant difference.4 Conclusion:Yiqi Qufeng, Xuan Bi Hua Yin Formula can inhibit c-Jun, c-fos gene expression and inhibition of AP-1, and further inhibit IL-4, IL-5 and IL-13 production. Yiqi Qufeng, Xuan Bi Hua Yin Formula can improve the CARAS guinea pig model of symptoms and lung pathology and the nasal mucosa pathological syndrome, inhibit the eosinophil eosinophil accumulation. While the existence of the relationship between therapeutic effects of Yiqi Qufeng, Xuan Bi Hua Yin Formula, namely the high dose with low dose, more conducive to pathological improvement, inflammatory factors and gene low expression, protein content reduced.