| Objective: Otitis media is a middle ear inflammatory disease with infection as the main cause,middle ear mucosal injury as the main pathological manifestation,and progressive hearing loss,tinnitus and earache as the main clinical manifestations.It is the most common disease which affected middle ear mucosa's function.Trefoil factor family(TFF)is closely related to respiratory system and plays an important role in the protection and repair of respiratory mucosa.However,the role of TFF in the pathogenesis of chronic otitis media and its mechanism have not been reported.In this study,the expression of TFF1 and TFF3 proteins in the development of otitis media in rats were detected by building an animal model of otitis media in rats,and the relationship between the expression of TFF1 and TFF3 proteins and the expression of inflammatory factors in the middle ear mucosa was studied.The possible role of TFF1 and TFF3 proteins in the pathogenesis of otitis media was discussed.Methods: In the first part of the study,researchers bought 25 healthy adult male SD rats with the same age,in addition,50 ul type 3 streptococcus pneumoniae suspension was injected with bilateral auditory vacuoles to construct the otitis media model in rats.All of SD rats were randomly divided into 5 groups: sham operation control group,model 2-week group,model 4-week group,model 8-week group and model 12-week group.We utilized HE staining to observe the morphological changes of rat middle ear mucosal cells at different stages.The expression of TFF1 and TFF3 proteins in rat middle ear mucosa at different stages was detected by immunohistochemistry and Western blot.In the second part of this study,10 SD rats were randomly divided into 2 groups: sham operation control group and otitis media model group.The rat model of otitis media was constructed with the same modeling method as the first part.Two groups of rat middle ear mucosal tissue samples were collected,and the concentrations of TFF1 and TFF3 proteins,TNF-?,IL-6 and IL-8 in the tissues were detected by ELISA.Results: In the model group,collagen fiber aggregation,the fibrous tissue proliferation and vascularization were observed in HE staining of rat middle ear mucosa.Immunohistochemical staining showed that TFF1 and TFF3 proteins were expressed in the middle ear mucosal tissues of rats in the normal group and increased in the middle ear mucosal tissues of rats in the model group,mainly expressed in the cytoplasm of epithelial cells and fibroblasts.Western blot detection showed comparing with the sham operation control group,the expression levels of TFF1 and TFF3 proteins in the middle ear mucosal tissues of rats in the model group increased with the development of the disease,then decreased slightly at 8 weeks of modeling,and then increased,reaching a peak at 12 weeks of modeling.ELISA detection showed that the concentrations of TFF1,TFF3 proteins,TNF-?,IL-6 and IL-8 which was in the middle ear mucosa of otitis media group were significantly higher than those of the control group(P<0.05).Correlation analysis showed that the level of TFF1 protein in otitis media rats was positively correlated with the level of IL-6 and IL-8(P<0.05),but not with the level of TNF-?.The level of TFF3 protein was positively correlated with the levels of TNF-??IL-6 and IL-8(P<0.05).Conclusion: The expression levels of TFF1 and TFF3 proteins in otitis media rats were significantly increased,and the levels of TNF-?,IL-6 and IL-8 inflammatory factors in otitis media rats were significantly correlated with the levels of TFF3 proteins,beside,the levels of IL-6 and IL-8 were significantly correlated with the levels of TFF1 proteins.Thus,the levels of TFF1 and TFF3 proteins in the tissues can reflect the activity degree of the mucosal inflammation of otitis media in rats,and have a good evaluation efficacy for the inflammatory progress of otitis media.Significance: This topic which built the rat animal model of otitis media firstly discussed the TFF1 and TFF3 protein expressed in rats' middle ear inflammation control theory besides related in vitro experimental methods,and made clear the TFF1 and TFF3 protein could be regard as an important regulatory factors in middle ear mucosa inflammation,which involved in the occurrence and development of the middle ear inflammation.The results of the study will enrich the theoretical understanding of the pathogenesis of otitis media,provide new ideas for targeted based on the treatment of otitis media as well. |
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