The Expression Of STAT6and IL-4in Nasal Mucosa Of Guinea Pig Allergic Rhinitis Models By Different Prophylactic Time

Background and ObjectiveAllergic rhinitis (AR) is the chronic inflammatory disease of nasal mucus triggered by IgE antibodies involved, mast cells release mediators (mainly include histamine, etc.) at the beginning, and the combined action of multiple immuno-active cells and cytokines.20years ago, under the base of former experiment, Ciprandi G proposed the term of allergic rhinitis minimal persistent inflammation (hereafter referred to as AR-MPI), it is that there is without any symptoms when patients suffered AR under low allergen, but the nasal mucosa inflammation have been exist, and he also put forward the method of prevention treatment, that is giving previous medicine therapy before symptoms happening, so it will reduce the incidence of complication and improve the life quality of the patients. Although it was not accepted widely, people get enlightenment of AR treatment from it and researches related AR with preventive treatments have never been stopped. The establishment of animal model of AR-MPI gives a opening of the research. This dissertation is to observe the nasal mucosa pathological changes and expressions of IL-4and STAT6during guinea pig experiment with previous treatment with glucocorticoid at the time of one week, two weeks, four weeks to explore the AR previous treatment effect in different time with medicine treatment.Methods120health Hartley guinea pigs, without gender limited and the weights are between200g～300g, they were randomly divided into six groups:A. Control group; B. AR without intervention; C. AR-MPI without intervention; D. AR-MPI with glucocorticoid treatment ahead of one week; E. AR-MPI with glucocorticoid treatment ahead of two weeks; F. AR-MPI with glucocorticoid treatment ahead of four weeks, twenty in each group, Groups B、C、D、E and F were treated with ovalbumin(OVA) by basic and strengthen sensitization, and then did the part intranasal excitation to establish the guinea pig allergic rhinitis and allergic rhinitis MPI models, then group B treated with1%OVA, groups C,D,E,F of allergic rhinitis were treated by0.01%OVA in nasal and came out AR-MPI animal models. Group A were treated with normal saline in place of OVA, group D, E, F were treated with Fluticasone Propionate Nasal Spray (hereafter referred to as FP) as prevention treatment group, after the establishment of animal models, used FP(50μg/side/time) treatment twice per day, treated them with one week, two weeks and four weeks. Observed the behavioral changes and nasal mucosa got from killed guinea pigs after last treatment, used immunofluorescence technique localization to check STAT6and IL-4in nasal mucosa of guinea pigs in different group.Results1. Animal behavior observationThe superposition method of scoring was adopted to evaluate the guinea pig model, total score more than5shown that AR models are established successfully, to judge successful AR-MPI model in accordance with pathology of nasal mucosal. The treatment in different time, after the treatment of1%OVA, group A, E, F didn’t appear typical AR symptoms, Group B, C, D appeared typical symptoms of AR, the difference of group A, E, F is lass than0.05, and group B, C, D were same, there are without statistically significant, the difference between group A, E, F and B, C, D had statistically significant (P<0.05).2. The test result of nasal mucosa pathology After establishment of the animal models, compared the eosinophil (hereafter referred to as EOS) of nasal mucosa in different group, group B is more than group A, and the difference have statistically significant (P<0.05); compared with A, B, group C, D, E, F are less, the difference had statistically significant (P<0.05). it showed that the model were successful.After the FP treatment of topical nasal with1%OVA, the nasal mucosa of guinea pigs of group A had no obvious changes, group B appeared serious tissue remolding, Group C, D appeared a lot of EOS, group E, F were completed, group B had difference with others (P<0.05), group A, E, F were without difference(P>0.05), group C and D had no difference (P>0.05), group A, C, D had difference(P<0.05).3. The test result of immunohistochemical of STAT6and IL-4STAT6and IL-4could be detected in nasal mucosa of normal and AR guinea pig, the expression were basically conformity, mostly concentrated in the cytoplasm of sub-epithelial infiltrative cells.After the establishment of animal models, the expression of IL-4and STAT6were as follows:Group A appeared brown granules; group B was more than A; brown granules distributed on whole of group C, D, E, F, the difference of groups had no statistically significant (P>0.05), the difference between group A, B and C, D, E, F had statistically significant (P<0.05).After FP treatment with1%OVA again, the expression of IL-4and STAT6were as follows:Group A had no obvious change; group B was serious, group C, D appeared by group, group E, F distributed by dot. Compared the difference, Group B had difference with others (P<0.05), Group A, E, F had no difference (P>0.05), group C, D were without difference (P>0.05), group A, C, D had difference (P<0.05).Take group B as sample, after establishment of AR model, compared the gray value, they had clear correlations (r=0.960, p<0.05).Conclusions1. The expression of IL-4and STAT6were found in the nasal mucosa of both normal guinea pigs and allergic rhinitis guinea pigs. Both of them have correlation and STAT6could be used as index of estimating severity of AR.2. FP treatment can down regulate the expression of STAT6and IL-4; it is possible that it is one of the methods for AR treatment.3.Two weeks is the best prophylactic time to prevent guinea pig AR model previously, and can relieve the inflammation of AR after one week.