Study Of Regulation Of IL-4 And IL-13 Membrane Receptor Expression On STAT-6 Signaling Pathway To Investigate The Mechanism Of Pingxiao Granules In Treating Bronchial Asthma Rat Model

Background:Pingxiao granule is the experience of tutor Professor Shi Suofang in the treatment of bronchial asthma.The early clinical observation shows that the curative effect is accurate,but its mechanism is still not clear.This study is based on the expression of IL-4R and IL-13R?1,IL-13R?2 and then regulate the STAT-6 signaling pathway,and to explore the mechanism of the treatment of bronchial asthma.Objective:This experimental study is to elucidate the Pingxiao granule by IL-4,IL-13 membrane receptor expression mechanism of STAT-6 signaling pathway in the treatment of bronchial asthma.At the same time,it provides molecular biological evidence for the effective treatment of asthma.Methods:60 SD rats?clean grade?,are randomly divided into five groups:blank group?C-G?,model group?M-G?,DXM group?D-G?,Ping Xiao low dose group?L-G?,Ping Xiao high dose group?H-G?,twelve rats in each group,half male and half female.In addition to the blank group,rats of each group by intraperitoneal injection of suspension solution?OVA+AlO₃H₃?and inactivated pertussis sensitization,and OVA inhalation induced asthma model;the blank group by intraperitoneal injection of the same volume of physiological salineinhalation of distilled water.After the establishment of the model,the experimental groups are given corresponding drug treatment;the blank group is given distilled water as control.During the course of the experiment,the symptoms of rats are observed closely,and the scores are evaluated by asthma behavior.After 14 days of drug intervention,rats are killed,serum and lung tissue of rats are detected:1)to observe the pathological changes of lung tissue by H.E staining;2)Giemsa staining under optical microscope observation of lung tissue eosinophilia;3)ELISA method is used to detect the changes of serum IL-4,IL-12 and IL-13 concentrations;4)the expression of pSTAT-6 in lung tissue is detected by western blot analysis;RT-PCR is used to detect the expression of IL-4Ra,IL-13 Ral/a2 of lung tissue.Results:1)In this study,OVA,Al?OH?₃ and Bordetella pertussis are combined to induce sensitization and induce asthma.The model is fit to the basic characteristics of asthma animal model,and the experimental model is established.2)HE staining:There is no significant change in lung in blank group.In the model group,the bronchial wall is significantly thickened,and the exudate is present in the lumen.There are a lot of inflammatory cells infiltration,fibrous connective tissue proliferation,and so on.In the other three groups,there is a slight thickening of bronchial smooth muscle and interstitial infiltration of inflammatory cells.3)Giemsa staining:there is no obvious abnormality in the lung tissue of the blank group,and there is a large number of eosinophils infiltration in the model group.In the other groups,there is a small amount of eosinophil infiltration.4)ELISA detection:IL-4 level:there is no significant difference between the five groups?P>0.05?.IL-13 level:compared with the model group,the IL-13 level is significantly lower?P<0.05?.However,the level of IL-13 in the model group is the same as that in the other groups,and there is no significant difference between the blank group and the other groups.IL-12:the OD value of almost all is negative,and has no significance.5)RT-PCR detection:IL-4R mRNA:the model group is significantly higher than the other four groups?P<0.05?,and the model group and dexamethasone group,Chinese medicine group and control group had significant difference?p<0.01?;and the blank group is significantly lower than the model group and the low,and the difference is significant?P<0.01?,but when compared with the other two groups level,no statistical significance;in the high group and dexamethasone group and low group had no significant difference.IL-13Ral mRNA:the model group is significantly higher than the other four groups,the difference is statistically significant?p<0.05?,and blank group,dexamethasone group and high group compared with the model group,there is significant difference?p<0.01?;control group is significantly lower than that in the model group,high and low,the difference is statistically significant?p<0.05?.And the model,the low group and the control group there is significant difference?p<0.01?;the level of M group and high group,no significant difference?P>0.05?,but with low group than the difference is statistically significant?p<0.05?;traditional Chinese medicine or the level of the two groups,no statistical significance?P>0.05?.IL-13Ra2 mRNA:the level of model group is significantly higher than the other four groups is statistically significant?P<0.05?,and model group and dexamethasone group,high group and blank group had significant difference?p<0.01?;control group compared with the model group,the level is low,and the difference is significant?P<0.01?,but with the other three groups the same level,no significant difference?P>0.05?.The levels of the three groups are similar?p>0.05?.Conclusion:the pathological changes of lung tissue and the related inflammatory indexes are improved after the treatment of asthma and dexamethasone.Ping Zhu granule can inhibit the phosphorylation of STAT-6 and reduce the release of Th2 type cytokines by inhibiting the membrane receptor IL-4R and IL-13R alpha 2 of IL-4 and IL-13,so as to make the inflammatory response to Th1.This may be one of the mechanisms for treating allergic asthma.