The Role Of CCL8-Mediated STAT3 Signaling Pathway In The Pathogenesis Of Allergic Rhinitis

Objective To study the expression of CC Motif Chemokine Ligand 8(CCL8)in Allergic rhinitis(AR)mice and AR human nasal mucosa epithelial cells,and to explore the role of CCL8 in the pathogenesis of AR.The possible pathwaysinvolved in AR could be preliminarily explored to provide the theoretical basis for its prevention and treatment.Method 1.Animal experiments: 6 mice were successfully constructed as AR model group through basic sensitization and local stimulation,6normal mice were used as blank control group.The mice in the model group and the control group were observed for 30 minutes after being challenged with normal saline and OVA.The number of sneezing,the degree of nasal itching and the amount of nasal secretions in the two groups of mice were observed and recorded,and their behavioral performance was evaluated by scoring.HE was used to observe the infiltration of eosinophils in the nasal mucosa of two groups of mice.The serum of the two groups of mice was collected,and the serum IgE level was detected by ELISA.Two groups of mouse nasal mucosa tissues were taken,mice total RNA was extracted,and RNA samples were preliminarily quantified by NanoDrop 2000;The RNA sample concentrations were accurately quantified by Aglient 2100.After the total amount,concentration,integrity,and purity of RNA met the standards,paired-end PE150 sequencing was performed based on the IlluminaHiseq platform,with a data volume of 12 G per sample,followed by data quality control and bioinformatics analysis,selecting CCL8 gene,which was significantly rise in allergic diseases.2.Cell experiment part: culture human nasal mucosa epithelial cell line;use Der p1 to stimulate the nasal mucosa epithelial cell modeling,RT-qPCR was used to detect the expression levels of IL-4 and IL-13 levels.To determine whether the modeling was successful.RT-qPCR and Western blot were used to detect the expression of CCL8.In order to explore the possible mechanism of CCL8 involved in AR immune regulation,the CCL8 gene was transfected into human nasal mucosa epithelial cells by lipofectamine 2000.A cell model with high expression of CCL8 was established,and the gene expression levels of IL-4,IL-13,CCL8 and STAT3 were detected by RTqPCR;the protein of CCL8 and STAT3 were detected by Western blot.Results 1.Animal experiments: Through the established AR mouse model,it was found that the degree of nasal itching,the number of sneezes,the amount of nasal secretions,the number of eosinophils in the nasal mucosa,the level of serum IgE,and the expression of CCL8 gene were significantly different in the model group.higher than the blank control group,2.Cell experiment part: By establishing AR human nasal mucosa epithelial cell line,it was found that the expressions of IL-4,IL-13,CCL8 and STAT3 were all rise in AR human nasal mucosa epithelial cells compared with normal nasal mucosa epithelial cells.After transient transfection of CCL8 DNA into human nasal mucosal cells,STAT3 mRNA also increased.Conclusion The expression of CCL8 is increased in AR mice and AR human nasal mucosa epithelial cells,and CCL8 may participate in the regulation of AR immune pathway by regulating STAT3 signaling pathway.STAT3 signaling pathway may be a new way of AR targeted therapy in the future...