| 1. Background and objectiveAllergic rhinitis (AR) is noninfectious hypersensitivity reaction of Type I which isthe chronic inflammatory disease of nasal mucus triggered by IgE antibodies involved,mast cells release mediators, and it would cause the release of inflammatorytransmitters、neuropeptide and cytokine after hypoallergenic contact with specificityperson. Some reports indicated that the ratio of allergic rhinitis is about10%-25%inglobal, but the ratio in Chian is15%-22%approximately. Allergic rhinitis is one ofcommon dieases in the clinical ENT department, but the clinical effects of it wasmultiplicate as well. Until now, the mechanism of AR of how it happen is unclear, butthe clinical research is going on and had indicated that AR may be concern about theenvironment and heredity. People known that the pathogenesis of AR is morecomplex. Currently, the accepted theory of pathogenesis is Th1/Th2imbalance theory,the specific expression of cytokines secreted by its uneven due. Intercellular adhesionmolecule (ICAM-1) and interleukin-12(IL-12) are two of important chemokine inmediated inflammatory reaction. In recent years, some scholars have proposed that itis much better to joint nasal corticosteroid and nasal vasoconstrictor to relieve ARsymptoms than the simple application of nasal corticosteroids. This study was designed to detect the changes intercellular adhesion molecule1and interleukin-12innasal mucosa by immunohistochemical method and to investigate the influence ofnasal corticosteroid and nasal vasoconstrictor on animal model of allergic rhinitis inrats, in order to provid the theoretical basis in animal experiment and molecular levelfor optimizing the prevention and treatment of allergic rhinitis.2. Materials and Methods2.1Experimental animals and groupingA total of sixty Sprague-Dawley (SD) rats, male or female,4-6weeks old and180-220weight, were included in this experiment. All of the animals were sensitizedwith ovalbumin(OVA)(OVA10mg, aluminum hydroxide gel20mg and PBS liquid1ml) to established the rat model and then divided into three groups: AR shame group(group A), xylometazoline hydrochloride treatment group (group B), xylometazolinehydrochloride and mometasone furoate combined treatment group (group C). GroupA was treated with saline, group B was treated with xylometazoline hydrochlorideintervention, and group C was treated with xylometazoline hydrochloride joint nasalcorticosteroids intervention.2.2Animal modelsAll of the experimental animals were treated with ovalbumin(OVA) by basicsensitization to established the AR rat model. First of all, using the allergensuspension which contains10mg OVA, aluminum hydroxide gel20mg to basissensitized by intraperitoneal injection, every other day, a total of seven times.Followed by the strengthen sensitization: using a concentration of5%OVAsuspension50ul to stimulate the bilateral intranasal once daily for two weeks. Torecording and score the nasal symptoms, from the first time that using the dose, forexample the nasal itching (sensitized animals with paws scratching nose), sneezing,nasal discharge, etc. At the end of treatment, killed the rats by decapitation, taken thenasal mucosa and fixed in10%formalin.2.3Score standards of nasal symptomThe score standards of nasal symptom were as follows:(1)Nasal itching:0pointswithout scratching nose; mild nose grab1point;2points for frequent nose grab; grabthe nose more than3points.(2) Sneeze: No sneeze0points;1point sneezing1-3; 4-10to2points;11more than3point.(3) Runny nose: No nose is0; the flow to theanterior nares of1points;2points beyond the front nose; tears streaming sounded forthe3points. When recording the score, use the superposition method to finish it. Thepoints greater than five indicates that the AR rat model is successful.2.4Histopathological observationThe organization will placed in10%formalin solution soak to fixed,paraffin-embedded sections under observation, using HE staining and observed in10×20optical microscope.2.4ICAM-1and IL-12protein expression in nasal tissueOperate by the immunohistochemical methods to accordance with the kitinstructions. The positive of ICAM-1、IL-12expression was shown brown particlesunder the microscope. Select five horizons of each film randomly for imageacquisition. Application Biosens Digital Imaging System v1.6to calculate the meanoptical density of the collected photographs of immunohistochemical staining weremeasured. Repeated three times and averaged.2.5Statistical analysisThe SPSS18.0software was used in this study for statistical analysis. Data werepresented as mean±standard deviation (S.D.). One-way ANOVA was conductedacross all groups first followed by a Bonferroni post-hoc correction between all groupcomparisons. Significance was accepted at p﹤0.05.3. Results3.1Animal behavior observationSuperposition method of scoring was adopted to evaluate the AR rat model.Total score more than5indicated that AR models are established successfully, tojudge successful AR model in accordance with pathology of nasal mucosal. Afterusing the drug therapy, three groups showed that typical symptoms and thedifference was obviously（P<0.05）. Compared with group B, the decrease of groupC was much more significant （P<0.05）.3.2The result of nasal mucosa pathologyCompared the eosinophil (hereafter referred to as EOS) of nasal mucosa indifferent group showed that group A is more than other groups, and the differences have statistically significant (P<0.05); compared with group B, there is present muchfewer in group C, the difference had statistically significant (P<0.05).3.3The test result of immunohistochemical of ICAM-1and IL-12ICAM-1is usuarally present in the cytoplasm of epithelial cells, such aseosinophils, basophils eosinophilic in the nasal mucosa of rats. The results indicatedthat there were a great quantity of positive cells in nasal mucosa of group A, while thepositive cells in group B and C had a mild expression. The group A had significantdifference compared with the group B and C（P<0.05）. Compared with group B, thereis present much fewer in group C, which the difference had statistically significant(P<0.05). IL-12is mainly present in mucosal epithelium and glandular epithelium ofnasal mucosa, showed brown yellow. The positive cells had sporadic expression ingroup A and a large quantity expression in group B and C. The expression of positivecells was significant in group B and C, compared with group A. While group B and Chad obvious difference compared with each other as well（P<0.05）.4. ConclusionsXylometazoline hydrochloride combined mometasone furoate treatment cansignificantly reduce the symptoms of AR rats.The expression of ICAM-1and IL-12plays an important role in allergic rhinitis;the mechanism may be associated with reduced expression of ICAM-1in nasalmucosa, increased expression of IL-12on the nasal mucosa, thus reducing nasalinflammation.It is much better to joint Xylometazoline hydrochloride and mometasone furoateto relieve AR symptoms than the simple application of nasal corticosteroids. |
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