| Objective:asthma is an immune-inflammatory disease in which T lymphocyte-associated airway inflammation plays an important role.Airway inflammation is characterized by Th2 cell activation,eosinophilic infiltration of EOS and increased IgE.Thl/Th2 cell imbalance plays an important role in the pathogenesis of asthma.IL-27 plays an important role in the pathogenesis of asthma.IL-27 can activate the STAT1 pathway of early activated T cells,thereby promoting Thl type immune response and inhibiting Th2 immune response.STAT1 phosphorylation disorder in airway of bronchial asthma leads to impaired function of I1-27 inhibiting Th2 cells,leading to imbalance of Thl/Th2 and inducing and aggravating asthma.Therefore,increasing the phosphorylation level of STAT1,enhancing the inhibitory effect of IL-27 pathway on Th2 cells,and restoring the airway Thl/Th2 immune balance in asthma patients provide a new pathway and target for asthma treatment.Xiaochuanning granule has significant clinical efficacy.Previous studies have confirmed that Xiaochuanning granule can inhibit the activation of T cells in asthma model mice and promote Thl/Th2 balance.Based on the regulatory effect of IL-27 pathway on Th cell immune balance,we propose the scientific hypothesis that asthmatic Thl/Th2 immune imbalance and airway inflammatory response can be regulated by Xiaochuanning granule through regulating STAT1 pathway.This project based on the previous research work,by Xiaochuanning granules intervention in asthma rats model,compared with the rest of the group,adopting bronchoalveolar lavage,immunohistochemical and WB,technology and method,respectively,the model of bronchial lung tissue,bronchoalveolar lavage fluid,such as in-depth study,reveal Xiaochuanning granules in the role of IL-27 access control targets,to clarify Xiaochuanning granules by STAT1 pathways regulating Thl/Th2 immune balance mechanism of bronchial asthma and provide objective experimental basis for its clinical application.Methods:based on the previous work,male SD rats were randomly divided into normal group,model group,IL-27 prevention group,IL-27 treatment group,asthmatic Xiaochuanning granules group and asthmatic Xiaochuanning granules plus STAT1 inhibitor group.Adopt the method of egg protein mixed sensitization,copy the rat model of asthma,Xiaochuanning granules and IL-27 to asthma intervention in rats,compared with the rest of the group,observe the relevant indexes in serum and BALF,lung tissue concentration,positive cells expression and the change of protein expression,study Xiaochuanning grain IL-27-STAT1 pathways through the regulation of immune inflammation of asthma rats imbalances.Results:(1)compared with the normal group,a large number of inflammatory cells infiltrated into the bronchial,perivascular and lumen,macrophages,eosinophils and lymphocytes were observed in the lung tissue sections of the model group.Edema,degeneration and abscission of bronchial mucosa epithelium;Capillary hyperemia,bronchioles and terminal bronchioles can be seen mucous plug,bronchioles and small vessel wall slightly thickened;The alveolar cavity fusion is enlarged,the alveolar wall is partially thickened,and the alveolar epithelial cells are increased.The other groups showed less inflammatory damage than the model group.(2)immunohistochemical results showed that compared with the normal group,the level of IFN-y and STAT1 in the model group decreased(P<0.01),and the level of IL-4 increased(P<0.01).Compared with the model group,the levels of IFN-y and STAT1 were increased in the IL-27 prevention group,the IL-27 treatment group and the Xiaochuanning group(all P<0.01),and the levels of IL-4 were decreased(all P<0.01).Compared with the model group,the level of IFN-y was increased(P<0.01),the level of IL-4 was decreased(P<0.01),and the level of STAT1 was slightly increased in the Xiaochuanning plus STAT1 inhibitor group,but the difference was not statistically significant.It indicates that the model group of rats has Th2 immune response hyperactivity,Thl immune response is weakened,and the rat airway and lung tissue have obvious inflammatory reaction.Compared with the model group,the infiltration of inflammatory cells and airway secretions in the lung tissues of the IL-27 prevention group and the treatment group,the asthmatic Xiaochuanning group and the asthmatic Xiaochuanning plus STAT1 inhibitor group were reduced,suggesting that IL-27 and asthmatic Xiaochuanning granules have certain anti-inflammatory effects on asthmatic rats.The effect of IL-27 prevention group was better than that of treatment group.The effect of asthmatic Xiaochuanning group was better than that of asthmatic Xiaochuanning plus STAT1 inhibitor group,and the difference was statistically significant.(3)compared with the normal group,the eosinophil count and percentage in the model group were all increased(P<0.01).In the serum and BALF of the model rats,the expression levels of IL-4 and IFN-y were significantly decreased(all P<0.01),and the expression levels of IFN-y,STAT1 and Samd2 were significantly decreased(all P<0.05),and the expression levels of IL-4 were significantly increased(P<0.05).Compared with the model group,the percentage of eosinophils in the blood of the other treatment groups was decreased(all P<0.01).The serum levels of IL-4 and IFN-? were decreased and increased in BALF of the other groups(all P<0.05).The expression levels of IFN-y,STAT1 and Samd2 were significantly increased in the IL-27 prevention group and the asthmatic Xiaochuanning group(all P<0.05),and the expression levels of IL-4 were significantly decreased(all P<0.05).The IL-4 expression level in the lung tissues of the IL-27 treatment group was decreased(P<0.05),but the expression levels of IFN-?,STAT1 and Samd2 in the lung tissues showed no statistical significance compared with the model group.There was no statistically significant difference in the expression levels of various indicators between the asthmatic Xiaochuanning plus STAT1 inhibitor group and the model group.The effect of the IL-27 prevention group was better than that of the IL-27 treatment group(P<0.05).The effect of asthmatic Xiaochuanning group was better than that of asthmatic Xiaochuanning plus STAT1 inhibitor group(P<0.05).Conclusion:(1)both Xiaochuanning granule and IL-27 prevention group can promote the phosphorylation expression of STAT1 in rat lung tissue and activate the signal transduction pathway of STAT1 cells,so as to enhance Thl immune response and inhibit Th2 immune response,and thereby reduce the inflammatory response in the lung tissue of asthma-rats,while Xiaochuanning granule can block this effect by STAT1 inhibitor.Based on the above results,we speculated that the mechanism of asthmatic attack inhibition by Xiaochuanning granule may be related to the IL-27 regulatory pathway.(2)Xiaochuanning granules may activate the downstream STAT1 pathway through the IL-27 pathway,and then inhibit IL-4,increase IFN-? and restore Thl/Th2 cell balance by increasing the phosphorylation level of STAT1,so as to exert the inhibitory effect of airway inflammation. |
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