A Research On The Intervention Of Xingbi Gel In Allergic Rhinitis And Its Effect On Fyn-STAT5 Signaling Pathway

ObjectiveIn this study,guinea pigs with allergic rhinitis(AR)and rat nasal mucosa fibroblasts isolated in vitro were selected as the research object,and the role of Fyn-STAT5 signaling pathway in the pathogenesis of AR was analyzed by in vivo and in vitro combined research methods.And Xingbi Gel was used to clarify its mechanism of intervention in allergic rhinitis in order to establish the experimental foundation of improving the clinical efficacy of allergic rhinitis.Methods1.Experimental study on the intervention of Xingbi Gel in AR guinea pig model and its effect on Fyn-STAT5 signaling pathway.60 healthy and clean guinea pigs aged 3 months were selected,half male and half female,and were randomly divided into blank control group(group A,n = 15)and model group(n =45),and the latter received intraperitoneal injection of ovalbumin(OVA)and Al(OH)3 mixture to establish AR model.After the modeling,the subjects,according to their weight and sex,were divided into AR model group(group B),Xingbi Gel treatment group(group C)and budesonide nasal spray control group(group D)by stratified randomization.The administration began 2 days after the intraperitoneal injection,and both A,B groups received saline nasal drops while group C received 50?l of Xingbi Gel stoste and group D received 50?l of budesonide nasal spray stoste,3 times a day,continued for 11 days.When provocated,group A received 50?l of saline nasal drops,and group B,C,D received 2%OVA solution intranasally.The provocation began 4 days after the intraperitoneal injection,once every two days,5 times in total.The guinea pigs were executed as soon as the administration was over,and serum IL-10 and MMP10 levels of the arterial blood were measured by ELISA.The nasal mucosa of guinea pig was taken and its morphology was observed by HE staining,and immunohistochemical method was used to detect the expression of CD19 and CD23 protein in nasal mucosa.The expression of MCP-1,MMP-10 and TGF-?1 mRNA was detected by real-time PCR.Western blot was used to detect the expression of Fyn and STAT5 protein.2.Experimental study on the effect of Xingbi Gel on Fyn-STAT5 signaling pathway in nasal mucosa fibroblasts of AR rat model.10 healthy and clean rats aged 2 months were selected,half male and half female,weight 150g-250g,and were divided into normal group(n = 5)and model group(n = 5)by stratified randomization.Five NF-?B knockout rats aged 40 days were selected,including 3 females and 2 males,weight 130g-200g.The modeling methods of the model group and the NF-?B knockout AR group were the same as in vivo experiment.Bilateral nasal mucosa was separated after the rats were executed,and the fibroblasts were extracted to be purified by differential adhesion method.The morphology of the fibroblasts was observed from the morphological point of view,and the vimentin related antigen was detected by immunocytochemical method.Fibroblasts were divided into normal rat nasal fibroblasts group(group A),AR rat nasal fibroblasts group(group B),NF-?B knockout AR rat fibroblasts group(group C),NF-?B knockout + TGF-?1 group(group D),NF-?B knockout +Xingbi Gel group(group E)and NF-?B knockout + budesonide nasal spray group(group F).Cells in each group were intervened for 12 hours,24 hours and 48 hours respectively.After the intervention,cells were collected and the proteins were extracted,and then the expression level of Fyn and STAT5 protein in each group was detected by Western blot.Results1.The success rate of AR guinea pig model was 86.67%,6 experimental guinea pigs of 45 fell off,among which 2 died and 3 were excluded because their score was less than 5.2.AR guinea pig nasal mucosa HE staining showed eosinophil infiltration,both the Xingbi Gel and budesonide nasal spray intervention could significantly improve the nasal mucosa eosinophil infiltration(P<0.05)and inhibited the expression of CD 19 and CD23(P<0.05).There was no significant difference between the Xingbi Gel group and the budesonide nasal spray group(P>0.05).3.Both Xingbi Gel and budesonide nasal spray could intervene in AR guinea pigs to lower the levels of IL-10,MMP10,TGF-?1 MCP-1 and MMP10 mRNA in serum(P<0.05),which could also inhibit Fyn expression and up-regulate STAT5 expression(P<0.05).There was no significant difference between the Xingbi Gel group and the budesonide nasal spray group(P>0.05).4.After NF-?B gene knockout,the Fyn expression in AR rat nasal mucosa fibroblasts was significantly inhibited while the STAT5 expression was significantly up-regulated(P<0.05).5.After TGF-?1 intervention,the expression of Fyn in AR rat nasal mucosa fibroblasts was significantly inhibited while STATS expression was significantly up-regulated(P<0.05).6.Both Xingbi Gel group and the budesonide nasal spray group could significantly inhibit,in vitro,the signaling pathway of Fyn-STAT5 in AR rat nasal mucosal fibroblasts(P<0.05),and there was no significant difference between the two groups(P>0.05).Conclusions1.Xingbi Gel inhibit the activation and degranulation of mast cells by reducing the production of inflammatory mediators and cytokines as well as inhibiting the synthesis of IgE receptors,so as to treat AR.2.Xingbi Gel inhibit the activity of mast cells to reduce allergic reaction by inhibiting Fyn-STAT5 signaling pathway,which is also one of its mechanisms of treating AR.3.NF-?B and TGF-?1 participate in the activation of nasal fibroblasts by mediating Fyn-STAT5 signaling pathway,which has an inhibitory effect on this signaling pathway.4.Xingbi Gel has an inhibitory effect on nasal mucosa fibroblasts in vitro,and this inhibitory effect is achieved by modulating the Fyn-STAT5 signaling pathway,which inhibits the activation of nasal fibroblasts and reduces the incidence of AR.