Xinfeng Capsule Improved Submandibular Gland Saliva Secretion Function On Primary Sj(?)Gren Syndrome Mice Modle And EGR1-STAT3 Signal Ing Pathway

Objective: To explore the mechanism of saliva secretion function of XFC in the treatment of primary Sj(?)gren’s syndrome by establishing animal models of primary Sjogren syndrome(ESS),and to investigate whether the target gene EGR1 can be used as a biomarker in primary Sj(?)gren’s syndrome.Methods: Mouse salivary gland protein induced the establishment of an animal model of primary Sj(?)gren’s syndrome.C57BL/6 mice were randomly divided into normal group,model group,low-dose group,and high-dose group,with 10 animals in each group.After taking C57BL/6 mouse submandibular gland shearing and grinding,BCA protein was used for quantification.Emulsified to a concentration of 2.0 mg/ml with an equal volume of Freund’s complete adjuvant,mice are anesthetized and injected continuously into the abdominal and dorsal subcutaneous multi-point for 3weeks.In the fourth week,it was injected with Freund’s incomplete adjuvant emulsification.Mice that successfully selected models were gavaged six days a week using fresh air capsules for four consecutive weeks.During this period,the relevant measurement data of mouse weight and saliva volume were collected.After the gavage is completed,mouse submandibular gland tissue is collected for tissue sequencing and follow-up experiments.Pathological staining observed the pathological condition of mice;RNA-seq sequencing of mouse submandibular glands to identify differentially expressed genes.RT-q PCR detected the expression of submandibular gland M3 R and target genes in mice before and after the intervention of fresh air capsules.WB detected the expression of submandibular gland M3 R and target gene in mice before and after the intervention of fresh air capsule.Data analysis predicted and detected the downstream target of the target gene EGR1.Results:1.Successful establishment of a mouse model of Sjogren syndrome;Compared with the control group,mice with primary Sj(?)gren’s syndrome lost weight,had slightly dry coat color,and reduced saliva volume;2.The results of pathological staining of submandibular gland showed that mice with model group had obvious lymphocyte infiltration compared with the pathological staining of the control group,and the number and scope of infiltrates increased;3.RNA-seq sequencing revealed the differentially expressed gene of model group,then verified by RT-q PCR in mouse submandibular gland tissue,finally the highexpression differential gene EGR1 was selected for subsequent experiments;4.After the administration of Xinfeng Capsules,the saliva volume increased significantly;Pathological staining of the submandibular gland showed a significant reduction in the extent of lymphatic infiltration;5.Compared with the control group,M3 R expression in model group was reduced;While compared with the model group,the expression of M3 R in the high-dose group of Xinfeng Capsules,was significantly increased;6.Compared with the control group,the expression of the target gene EGR1 in model group was significantly increased,and it decreased after the administration of Xinfeng Capsules;6.STAT3,as the downstream target of EGR1 was predicted and verified in mouse submandibular gland tissue,and the difference was statistically significant.Conclusion:1.Xinfeng Capsules has a significant effect on the salivary secretion function of Sjogren’s syndrome model mice.2.EGR1 can be used as a biomarker and therapeutic target for primary Sj(?)gren’s syndrome;3.It was preliminarily explored that Xinfeng Capsules can affect saliva secretion and relieve dryness symptoms by regulating EGR1-STAT3 signaling pathway;...