Effect Of Bi Min Fang On AQP5 In Nasal Mucosa Of Rats Of Allergic Rhinitis With Lung And Spleen Qi Deficiency Syndrome Though NF-κB Pathway

ObjectiveSet up the rat model of allergic rhinitis with deficiency of lung and spleen qi syndrome,and intervene with Bi Min Fang,and then observed the effect of NF-κB pathway on the expression of Aquaporin 5(AQP5)in the nasal mucosa,to explore the mechanism of "Enhancing earth to supplement metal”method for preventing and treating allergic rhinitis.Methods1.The rats were given smoking,senna gavage,intraperitoneal injection of ovalbumin and aluminium hydroxide suspension,and nasal provocation to set up the rat model of AR with deficiency of lung and spleen qi syndrome.During the modeling period,the general condition and nasal symptoms of the rats were observed,the body weight,food intake and swimming load time were measured to evaluate the rat model of deficiency of lung and spleen qi syndrome.Serum IgE levels were measured by ELISA,nasal mucosa were observed by pathological HE staining,and calculated the behavioral scores to evaluate the animal model of AR.2.Establishing the rat model of AR with deficiency of lung and spleen qi syndrome again.The rats were administered with Bi Min Fang of low concentration(11.3g/kg/d)and high concentration(22.6g/kg/d)by gavage for two weeks and four weeks respectively.During the experiment,the general conditions of the rats were observed.The rats were sacrificed after the intervetion.The nasal mucosa tissue of each group were observed by pathological HE staining.And immunohistochemistry and Western blot were used to detect the expression of NF-κB and AQP5,the technology of reverse transcription-polymerase chain reaction(RT-PCR)was used to detect he expression of IL-1β,TNF-α,IL-6 and AQP5.Results1.The behavioral scores of the rats in AR model group and AR with deficiency of lung and spleen qi syndrome model group were greater than or equal to 5 points.And the HE pathological sections of nasal mucosal epithelial cells under light microscope showed inflammatory cell infiltration,cilia shedding,disorder,and severe structural breakdown.Compared with the normal group,the serum IgE levels of the two model groups were higher(P<0.05),and there was no significant difference between the groups.Compared with the normal group and the AR model group,the AR with deficiency of lung and spleen qi syndrome model group showed the signs of the deficiency of lung and spleen qi,such as low energy,less activity,yellow hair color and more shedding,thin fecal matter,diarrhea,and even anorectal prolapse.Meanwhile,the food intake,body mass,weight-bearing swimming time decreased significantly(P<0.05).There was no significant difference between the normal group and the AR model group in the above-mentioned index.2.Compared with the normal group,the expression of NF-κB protein,the expression of AQP5 protein and mRNA,and the expression of IL-1β,TNF-α and IL-6 mRNA in the two model groups were significantly increased(P<0.05);compared with the AR model group,the activated nuclear NF-κB protein expression,and the TNF-α and AQP5 mRNA expression were higher in the AR with deficiency of lung and spleen qi syndrome model group(P<0.05).Compared with the AR with deficiency of lung and spleen qi syndrome model group,pathological sections of nasal mucosa of these four Chinese medicine intervention groups showed that the mucosal epithelial structure were almost recovered without obvious inflammatory cell infiltration.Meanwhile,NF-κB protein expression,AQP5 protein and mRNA expression,IL-1β,TNF-α and IL-6 mRNA expression were significantly down-regulated(P<0.05);compared between these four groups,only the high-concentration four-week group showed lower nuclear NF-κB protein expression than the low-concentration two-week group.Conclusion1.The method of smoking,senna gavage,intraperitoneal injection of ovalbumin and aluminium hydroxide suspension,and nasal provocationcan can establish the rat model of AR with deficiency of lung and spleen qi syndrome.2.Chinese medicine Bimin Decoction of“Enhancing earth to supplement metal" method can treat AR and reduce the hypersecretion of glands,which probably via inhibiting NF-κB signal pathway and down-regulateing the expression of AQP5.