| Purpose:To explore the effect of traditional chinese medicine recipes Sanao Decoction on the oscillating frequency of eustachian tube(ET)in rats with otitis media with effusion(OME),and the regulation of aquaporin 1(AQP1)and goblet cell number,providing valuable theoretical basis for the clinical use of Sanao Decoction in the treatment of OME.Material and method:Fifty-5-week-old male SD rats with healthy middle ear were adopted.After one week of adaptation feeding,normal saline or artificial gastric juice were injected with auditory vesicle puncture at the same time for modeling.After the completion of auditory vesicle puncture injection,ear endoscope was used to observe the modeling immediately,and the liquid plane and clear liquid were visible in the tympanum,indicating that the auditory vesicle puncture injection was successful.The rats with middle ear cavity bleeding were excluded,and 24 hours after the modeling,the modeling situation was observed through the tympanic membrane.In the group injected with normal saline by listening to the bubble,the fluid level disappeared and the tympanic membrane was transparent;in the group injected with artificial gastric fluid,the tympanic membrane was yellow and amber,with the fluid level and slight congestion,indicating that the modeling was successful.The rats were divided into three groups: model group,low dose group and high dose group.The middle ear cavity effusion was observed after 6 days of traditional Chinese medicine intragastric administration.The rats without auditory vesicle puncture were set as control group and sacrificed on the 7th day,as well as the other rats in the tracheostomy group.The Eustachian tube mucociliary oscillating frequency was analyzed,respectively,and morphological observation and associated protein study were carried out.Results:1.Auditory vesicle puncture injection of artificial gastric fluid can successfully create OME model of rats.The success rate of modeling is high,simple and easy,and it can be a new method of modeling.2.The low dose of Sanao Decoction has no therapeutic effect on the effusion of OME middle ear cavity.High dose can promote the discharge of middle ear effusion,the cure rate is 75%.3.The high dose of Sanao Decoction has a certain protective effect on ET mucosal cilia in OME rats: The immunohistochemical results showed that the high dose of Sanao decoction can reduce the loss of ET mucosal cilia caused by OME.Scanning electron microscopy(SEM)showed that the tips of ET mucosa cilia were clustered in rat OME model.After high dose treatment,Sanao Decoction could relieve the phenomenon of ciliary apex clustering and restore the coordination of ciliary oscillation.High-speed video microscope results showed that the high dose of Sanao Decoction could improve the ciliary oscillation frequency,and the ciliary oscillation frequency of ET mucosa in the model group was significantly decreased compared with the control group(p < 0.05).Although the ciliary oscillation frequency did not return to the normal level after high dose treatment with traditional Chinese medicine,it was significantly increased compared with the model group(p < 0.05).These results indicated that high dose of Sanao Decoction could improve ET function and promote effusion excretion by increasing the oscillating frequency of ET mucosa cilia.4.Immunohistochemical results showed that AQP1 expression in the model group was significantly lower than that in the control group.High dose of Sanao Decoction could enhance the expression of AQP1,but it did not reach the level before injection,suggesting that Sanao Decoction might enhance the water-liquid metabolism and accelerate the absorption and excretion of effusion by increasing the expression of AQP1.5.On the 7th day,the number of ET goblet cells in model group increased significantly(p <0.05),and the volume of ET goblet cells in model group was smaller than that in control group.High dose of Sanao Decoction could reduce the number of ET goblet cells(p < 0.05),but there was no significant change in the volume of ET goblet cells in model group.HE staining results showed that glandular mucosa in the model group was significantly thickened compared with that in the control group(p < 0.05).After high-dose treatment with Sanao Decoction,mucosa in the model group was improved compared with that in the model group(p < 0.05),but it did not recover to the level before injection.Conclusion:At high dose,Sanao Decoction can restore the function of eustachian tube,accelerate the absorption and excretion of secretions in middle ear cavity,reduce excessive secretion of mucus,and shorten the course of disease by accelerating the frequency of ET cilium oscillation,promoting the coordination polarity of cilium oscillation,increasing the expression of AQP1,and inhibiting goblet cell proliferation.However,morphology and function did not return to normal level in a short time,which still needs to be further studied whether this is the reason why OME is prone to recurrence,long treatment period and slow recovery needs further research. |
