Effect Of Menthol On The Occurrence And Development Of Chronic Rhinosinusitis

Background:Chronic rhinosinusitis(CRS)is the most common refractory chronic disease,mainly manifested as nasal congestion,runny nose,headache and olfactory disorder.Due to its strong heterogeneity and complex pathogenesis,the current treatment methods still can not achieve very ideal results.Traditional Chinese medicine and related studies of traditional Chinese medicine generally believe that Mint has potential therapeutic value in inflammatory respiratory diseases such as allergic rhinitis and chronic sinusitis.In clinic,a large number of CRS patients require compound menthol nasal drops in the outpatient clinic to alleviate symptoms such as nasal congestion,dizziness and headache,but there is no Study on the mechanism of menthol and its receptor in the treatment of CRS.The biology of respiratory epithelial cells can play a "bridge" role between innate and adaptive airway mucosal immunity.We found that menthol receptor TRPM8 is highly expressed in nasal mucosa epithelium of patients with CRS,but the role of menthol and its receptor TRPM8 in the occurrence and development of CRS has not been clearly studied.Objective:This study will explore the effect of menthol stimulated TRPM8 on epithelial cell function and the role of menthol stimulated TRPM8 in the occurrence and development of CRS diseases,in order to provide new ideas for the treatment and prevention and control of CRS.Methods:1.Clinical relevance study: compound menthol nasal drops were used for normal people and CRS patients.(1)The patients with CRS were assessed by SNOT-22 scale to determine the severity of the disease and the main symptoms of the patients.(2)Detect the changes of nasal resistance of normal people and CRS patients before and after use,(4)Compare the effects of vas nasal patency of normal people and CRS patients before and after use,(3)Compare the changes of nasal volume of CRS patients before and after use through CT reconstruction nasal volume model,(5)Compare the changes of emotional dimensions of normal people and CRS patients before and after use.2.Cytological analysis of the correlation between TRPM8 and CRS:(1)Collect clinical samples,analyze the expression level and high expression site of TRPM8 in nasal mucosa of patients with CRS,and analyze the correlation between the relative quantification of TRPM8 and the severity of CRS.(2)Cell experiment: human nasal epithelial cells(cell lines)were treated with menthol and NF-κB pathway inhibitor.The transcriptional levels of TRPM8,IL-33 and TSLP were detected by RT-PCR,and NF-κB phosphorylated p65 protein expression level was detected by WB.3.Animal model analysis:(1)The mouse model was established in three groups.Group A was ns nasal drip,group B was ova modeling CRS,and group C was synchronized with menthol nasal drip on the basis of group B;(2)The behavioral manifestations of scratching and sneezing of CRS mice in each group were recorded and analyzed;(3)The CT condition of nasal sinuses and mucosal HE staining of mouse model were compared to compare the degree of inflammation;(4)The expression level of TRPM8 in nasal mucosa and the expression level of epithelial cytokines IL-33 and TSLP were detected by ELISA;(5)The changes of spontaneous activity of CRS mice before and after menthol nasal drip were detected by open field test.Results:1.The SNOT-22 score of CRS patients showed that the most serious symptom was nasal obstruction;The nasal resistance of normal population and CRS patients increased after intervention compared with that before intervention.The nasal resistance of CRS patients was greater 5 minutes after intervention than 1 minute after intervention;VAS scale showed that subjective nasal patency increased in both groups after intervention compared with that before intervention,and vas patency increased more significantly in patients with CRS;CT reconstruction of nasal space in patients with CRS showed that the nasal space was significantly reduced after using compound menthol nasal drops;The emotional dimensions of the two groups were improved to a certain extent after the intervention.2.Compared with the normal control group,the expression of TRPM8 in nasal mucosa of patients with CRS was significantly higher,and the expression of TRPM8 in epithelial cells of patients with CRS was significantly higher than that of the control group.There was a positive correlation between snot score and TRPM8 protein level in nasal mucosa of patients with CRS.The m RNA transcription levels of TRPM8,IL-33 and TSLP in human nasal epithelial cells stimulated by menthol increased,and the higher the concentration and the longer the stimulation time,the more obvious the effect;After menthol stimulation,the expression level of phosphorylated p65 protein in NF-κB pathway increased significantly,and this effect decreased after using NF-κB pathway inhibitor.3.The frequency of scratching nose and sneezing symptoms in the three groups was the most severe in group C,followed by group B,and almost asymptomatic in group A;CT of mouse nose and paranasal sinuses showed that the swelling degree of nasal mucosa in group C was the heaviest,followed by group B;He staining showed that the degree of nasal mucositis in group C was the most serious,followed by group B;The expression level of TRPM8 in mouse nasal mucosa was the highest in group C,followed by group B;The expression levels of IL-33 and TSLP in nasal lavage fluid were the highest in group C,followed by group B;The open field test showed that the spontaneous activity time and distance of CRS mice increased significantly after menthol stimulation.Conclusion:The use of compound menthol nasal drops will increase the nasal resistance and reduce the space of nasal cavity and paranasal sinus in patients with CRS,but it will increase the subjective feeling of nasal patency.This open subjective feeling may be the "illusion" of neurosensory or the result of emotional improvement;Menthol stimulated HNECs TRPM8 can activate NF-κB pathway to further improve the expression of IL-33 and TSLP;Simultaneously dropping menthol in CRS mouse model can aggravate nasal mucosal inflammation and increase the levels of TSLP and IL-33 in nasal lavage fluid.Menthol may improve the mood of CRS mice by stimulating trigeminal nerve.The use of menthol nasal drops in patients with CRS is at risk of aggravating the disease.