| BackgroundAllergic rhinitis is a chronic inflammatory disease of the airway,which is essentially an allergic disease involving a variety of cells(such as eosinophIL-s,mast cells,T lymphocytes,neutrophIL-s,epithelial cells,etc.),inflammatory mediators and cytokine networks.Rhinitis is a complex and refractory disease that endangering the health of all age groups.Its incidence is increasing year by year.It is one of the hotspots in current research.The treatment of allergic rhinitis with traditional Chinese medicine has a long history and its own characteristics.It has been recognized by people in the continuous medical practice after it has made up for the short time of anti allergy,easy to recur and many side effects.In addition,the external treatment of Chinese medicine has its unique advantages.It can not only achieve the therapeutic effect of internal medicine,but also play an important role in the prevention of rhinitis and remission of rhinitis.The majie cataplasm used in our research is a new external dosage form.Its composition includes ephedra,mustard seed,rhizoma corydalis,bitter almond and ginger juice.As a new preparation for rhinitis,we need to study its clinical efficacy,experimental animal efficacy and mechanism.Therefore,this topic will be carried out from four aspects:clinical case study,network pharmacology research,drug effect and immunoregulation mechanism of majie cataplasm,so as to evaluate the therapeutic effect of majie cataplasm and explain the mechanism of its action.Objective1.Through clinical collection of allergic rhinitis patients,observe the clinical curative effect of majie cataplasm after the intervention.2.Through network pharmacology research,we excavate the potential target of majie cataplasm and predict the path of majie cataplasm.3.Through the preparation of BN rhinitis model,the drug treatment was used to observe the pharmacodynamics effect of majie cataplasm on rhinitis animal model.4.Through the preparation of BN rat rhinitis model,the effects of majie cataplasm on MAPK pathway,TSLP,Th1/Th2 balance and mucous connexin were observed,and the mechanism of immunoregulation was explained.Method1.48 cases of allergic rhinitis with lung qi deficiency and cold type were collected.After 4 weeks of acupoint application on the back of majie cataplasm,the symptoms of sneezing,nasal itching,runny nose and nasal plug were observed according to the visual analogue scale,and the clinical efficacy of majie cataplasm was evaluated according to the standard of curative effect.2.The components of ephedra,almonds,white mustard,Rhizoma Corydalis,ginger,and corresponding targets were inquired through the TCMSP database,and the NCBI Gene database was used to inquire the target of rhinitis.The target gene of majie cataplasm was fused with rhinitis target gene,and the protein interaction network was constructed through database.The core target was extracted from the protein action network,and the interaction protein was enriched and analyzed by DAVID database,and the specific pathway of majie cataplasm was selected to regulate the effect of rhinitis.3.Using OVA sensitized BN rats to prepare the animal model of rhinitis,which were divided into 4 groups:blank group,model group,cetirizine hydrochloride group(17.5mg/kg/d),majie cataplasm acupoint application group(6.65g/kg/d),10 rats in each group.After the success of the model preparation,the drug intervention was 6 weeks.Behavioral changes were observed during the treatment.After the treatment,the whole blood eosinophil was counted by rayleigh giemsa staining,and the infiltration of mast cells was observed by toluidine blue staining,and the pathological changes of nasal mucosa were observed by HE staining,and the content of IgE and histamine in serum and lavage fluid was detected by ELISA.4.The expression of ERK,p38,TSLP,T-bet,GATA,E-cadherin,and β-catenin protein was detected by Western Blot;the number of Thl/Th2 cells in whole blood was detected by flow cytometry;the contents of IFN,IL-4,IL-5,IL-13 and TSLP were detected by ELISA.Result1.Clinical observation:with the extension of the intervention time of majie cataplasm,the sneezing,itching,nasal congestion and runny nose symptoms have been alleviated effectively.The total effective rate of disease was 75%,and the symptoms such as nasal cavity examination,nasal mucosa and secretion were relieved.The effective rate of TCM syndrome reached 79.17%,and the symptoms of lung qi deficiency and cold type were relieved with self sweating,fear of wind,cold and cough.2.Network pharmacology analysis results:by searching the TCMSP database,we find 498 human target proteins(131 ephedra,117 almonds.177 yercords,46 white mustard seeds,27 ginger),and 364 rhinitis related human genes.74 core targets were screened by the fusion of target protein by Cytoscape,which involved airway inflammation,signal transduction,immunization and reaction,material synthesis and metabolism,airway remodeling,gene inheritance and other functions.DAVID database was used to analyze the pathway enrichment,and the MAPK pathway was used as a potential regulatory pathway for majie cataplasm.3.Pharmacodynamic results:(1)behavioral evaluation:after 6 weeks of treatment,both cetirizine group and majie cataplasm group could reduce the symptoms of sneezing after stimulation.Compared with the model group,the decrease of majie cataplasm was significantly different from that of the model group.After 6 weeks of treatment,both cetirizine group and majie cataplasm group could reduce the symptoms of nasal flex after stimulation.Compared with the model group,the decrease of majie cataplasm group was significantly different.(2)the count of eosinophils in the whole blood:the number of eosinophils in the peripheral blood of OVA induced rhinitis rats was reduced by the majie cataplasm group and cetirizine group.Compared with the model group,the number of eosinophils in the peripheral blood of majie cataplasm group decreased significantly.(3)the HE staining of nasal mucosa showed that the nasal mucosa of the rats in the model group was disordered,the cilium was different,there were obvious shedding,the epithelial cells were in confusion,the goblet cells increased,around the submucosa,the glands and vessels are dilated and proliferated,with congestion and tissue edema,and infiltration of eosinophiils and other inflammatory cells,showing the characteristics of mucosal remodeling.The nasal mucosa of cetirizine group improved significantly compared with the model group,the mucosa was more complete and smooth,and the tissue edema was relieved,with only a small amount of inflammatory cell infiltration.Compared with the model group,the majie cataplasm group improved significantly,and the mucosa recovered completely and smoothly,with only slight edema and a small amount of inflammatory cell infiltration.(4)the nasal mucous membrane toluidine blue staining:the model group can see a large number of mast cells infiltration,some cell cytoplasm particles release to the cell membrane,cetirizine group and majie cataplasm group mast cells significantly decreased.(5)the content of serum IgE and histamine:majie cataplasm can reduce the content of serum IgE and histamine.Compared with the model group,the decrease of IgE in serum has significant difference,and the decrease of histamine has very significant difference,and there is no statistical difference compared with cetirizine.(6)the content of IgE and histamine in the lavage fluid:the content of IgE and histamine in the lavage liquid could be reduced by majie cataplasm.Compared with the model group,there was no statistical difference of IgE,and there was significant difference in histamine reduction,and there was no significant difference compared with cetirizine.4.The results of immune mechanism research:(1)flow cytometry results:majie cataplasm can increase the number of Thl cells,which is significantly higher than that of the model group.majie cataplasm can reduce the number of Th2 cells,and is significantly lower than that of the model group.(2)WB test results:majie cataplasm can reduce the protein expression of ERK,p38,TSLP and GATA in nasal mucosa,and improve the expression of T-bet,E-cadherin,and beta-catenin protein.(3)ELISA test results:majie cataplasm can reduce the contents of IL-4,IL-5,IL-13 and TSLP in serum,and increase the content of IFN.Conclusion1.Majie cataplasm has certain curative effect on allergic rhinitis which is lung qi deficiency and cold type,and provides a basis and reference for the experimental study of the mechanism of the drug.2.Network pharmacology analysis and prediction of the MAPK pathway play an important role in the treatment of rhinitis with majie cataplasm.3.Majie cataplasm can inhibit nasal and sneezing allergy symptoms of rhinitis animal model,and reduce the content of IgE and histamine by inhibiting the infIL-tration of inflammatory cells,and play a role in the treatment of rhinitis.4.By inhibiting the phosphorylation of ERK and p38 and the content of TSLP,and regulating the Thl/Th2 balance and the expression of related cytokines in the downstream of the immune system,majie cataplasm can protect the function of E-cadherin and beta-catenin to alleviate the local nasal mucosa inflammation. |
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