The Expression And Role Of Nods Like Receptors In The Nasal Mucosa Of Patients With Allergic Rhinitis

Background:Allergic rhinitis (AR) is a nasal allergic disease with widely incidence, and20-50%incidence reported in the literature,which seriously plagues people in living, learning and working. Its incidence involves both environmental and genetic aspects, and the main treatment for it is drug therapy. ARIA defines it as a disease difficult to cure, but can be long-term controled. Thus, there is a need to clarify the pathogenesis and explore new therapeutic measures. Infection factors for allergy controversy exists. Mainly reflected in the theory of the "hygiene hypothesis" proposed early infection protection, infection-inhibiting allergy, Aaby reported that in French children, during infants and young children, inoculated with Mycobacterium was able to inhibit the occurrence of allergy on the allergy and infection aggravated this two aspects. Therefore, the impact of the infection becomes one of the research directions of the disease.Microbial stimulation of the nasal mucosa causes the body to start innate immunity, mainly by the pathogen associated molecular patterns (pathogen-associated molecular patterns, PAMPs) and mucosal cells pattern recognition receptors (Pattern Recognition Receptor, PRR), PRR is present in the cell a wide variety of family of receptor proteins, they can identify the tag of pathogenic microbes (bacteria, virus, etc.) or a danger signal from the host itself, thus triggering the immune response. The study shows that, PRR involved in the pathogenesis of many diseases, including allergy, autoimmune diseases and tumor. Toll-like receptors locate at the cell surface and specific organelles, such as endocytosis lysosomes main identify extracellular microorganisms, allergens and thus to identify the cell surface in allergic reactions and play an important role, thus, there have been more research in this area. But some microbes can escape the PRR of cell membrane and invade into the cell. How to identify such pathogens and intracellular bacteria, causing researchers to keep on exploring. Later found Nod-like receptor (Nod like receptors, NLRs) mode with this function. In recent years, this newly found but the "old" model receptor makes positive role in the inflammatory response in the upper respiratory tract, including allergic reactions, and draws much attention. While,the research of Nod1, Nod2and Nalp3Nod-like receptor family about allergic rhinitis remains poorly understood.The NLRs can identify microbial molecules in the cytoplasm. Nod1identifies tiny structures dipeptide through the meso-DAP(diaminopimelic acid), which is a characteristic component of the majority of Gram-negative bacteria and some Gram-positive bacteria. While, Nod2detects tiny structures-muramyl dipeptide (muramyldipeptide MDP), which is the minimum bioactive peptidoglycan motif present in all Gram-positive and-negative bacteria cell wall. After detecting their respective ligands, Rip2(serine, threonine kinase Rip2), and then mitogen-activated protein kinase (MAPK) and nuclear factor (NF-kappaB) are activated and the synthesis of IL-1β is any further promoted; Nalp3belongs to PYD subfamily molecule, which is able to identify the bacterial RNA, uric acid crystals, ATP,certain bacterial toxins, and aluminum hydroxide (alum). After Nalp1b, and Cryopyrin/Nalp3identification, the synthesis of inflammation bodies (inflammasomes) is promoted, thereby it activates caspase1(caspase-1, also known as IL-1β-converting enzyme.),and generates mature IL-1β molecule or may promote apoptosis.The study found, the main main indoor allergen is house dust mite. How to identify the house dust mite allergens and ingredients? Pattern recognition receptors TLR plays an important role. Such as Der p1is identified by TLR2, LPS through TLR4identification, and it shows TLR is related with allergic rhinitis. While,TLR is distributed only within the endosome membrane and intracellular, for other pathogens, which can evade the cell membrane into the cytoplasm component, TLR can not recognize, but the NLR can identify, the existence of such components can be found in the structure of the house dust mite.So NLR is very critical in disease caused by house dust mites in the innate immune incidenceIt reported that (Jacquet et al) microbial components can be detected in HDM medium,including lipopolysaccharide LPS and b-glucan class, and these components seem as important as HDM real antigen component.,it causes Th2by the activation of pattern recognition receptors,, the polarization. But whether the adjuvant composition alone can activate innate immunity, and further research is needed. He highlighted that house dust mite skeleton proteins contained chitin, which can be detected by surface pattern recognition receptor TLR-2recognition, resulting in pro-inflammatory and pro-inflammatory media. Further literature [7] reported,description adjuvant chitosan (chitin-based polymeric sugar), and silica, asbestos, aluminum hydroxide, can activate NLRP3(i.e. NALP3) and make signal transduction by antigen-presenting cell. Pattern-recognition receptors NALP3may play a key role.in the house dust mite allergen,as it can take in signal transduction, but the mechanism needs further study.Through NOD1gene polymorphisms research,Hysi [9] found, susceptibility to Card4variability was associated with increasing risk of asthma. Intracellular pattern recognition NOD1could identify bacteria-specific products, which affected the incidence of asthma in children. Asthma and allergic rhinitis just like "one airway, one disease", NOD1gene polymorphism may be associated with allergic rhinitis.Therefore, NLRs have a certain influence on immune function, and a role in the incidence of allergic rhinitis.It may be reflected in:1.Perception of micro-organisms, and mediate inflammatory response.2.They can cause the release of Th2cytokines after activation, thereby increase or maintain the inflammatory state of allergic rhinitis.3.Perception adjuvant can enhance the immune response.4Gene polymorphism may be associated with allergic rhinitis. This study is to further investigate the role of the NLR the acceptors (Nod1, Nod2, Nalp3) in allergic rhinitis.The experimental results are reported below.ObjectiveTo explore role of Nods (nucleotide-binding oligomerization domain Nod Like receptors) kind of pattern recognition receptors (PRR) in inferior turbinate in patients with allergic rhinitis and nasal polyps mucosa, deviated nasal septum.To compare tissue homogenate IL-4, IL-6, IL-10, IFN-r expression among allergic rhinitis, nasal septum deviation and nasal polyps. To explore the relationship between Nod1, Nod2, Nalp3and cytokine. To analyze Nod1expression changes of peripheral blood mononuclear cells after desensitization therapy.to learn change of IL-4, IL-6, IL-10, IFN-r in serum, nasal secretions after desensitization therapy. and relationship between Nod1expression change and cytokine change.MethodsDuring September2011and September2012, outpatient patients with allergic rhinitis (AR) in Zhujiang Hospital of Southern Medical University were recruited according to AIRA2010(based on clinical manifestations、nasal inspection, the skin prick experimental results or serum allergen-specific IgE detection during) into the group; deviated nasal septum alone orthotics patients as control group1, chronic sinusitis with nasal polyps (CRSw NP chronic rhinosinusitis with nasal polyps) hospitalized patients as control group2, according to EPOS2007Guide standards included in the study group. This study was approved by the ethics committee of Zhujiang Hospital of the Southern Medical University and informed consent of all subjects.Respectively, using real-time RT-PCR, western-blot and immunohistochemistry to analyze mRNA and protein of Nod1, Nod2of NALP3in the turbinate mucosa of patients with allergic rhinitis, nasal septum deviation nasal mucosa of patients with decompensated side, nasal polyp mucosal. using western-blot to learn Nod1expression changes in peripheral blood mononuclear cells after the treatment of desensitization. Level ofIL-4, IL-6, IL-10, IFN-r of allergic rhinitis, nasal septum deviation nasal mucosa, nasal polyps tissue homogenate were measured by ELISA,also with level of IL-4, IL-6, IL-10, IFN-r in serum and nasal secretions after desensitization treatment.Result1.Real-Time RT-PCR:mRNA expression of Nod1was decreased in mucosa of allergic rhinitis compared to normal control of the inferior turbinate mucosa and nasal polyps (P=0.31, P=0.00), NALP3mRNA expression was increased compared to normal control of the inferior turbinate mucosa, nose polyps (P=0.01, P=0.01), the Nod2mRNA expression among three groups showed no significant difference, the difference was not statistically significant (P=0.06, P=0.10).2.western-Blot results suggest that:Nod1expression of mucosa in allergic rhinitis was lower than the control inferior turbinate mucosa and nasal polyps group (both P=0.00),while NALP3was higher than normal control inferior turbinate mucosa, nasal polyps (P=0.00), Nod2among the three groups was not statistically significant (P=0.52, P=0.29).3.1mmunohistochemistry showed that the three modes of receptors in the three groups were expressed, they were mainly expressed in epithelial cells, glandular epithelial and inflammatory cells (e.g.plasma cells, eosinophils, neutrophils), allergic rhinitis mucosal tissue. Nod1expression was lower compared to normal control of the inferior turbinate mucosa, nasal polyps (P=0.01, P=0.00), NALP3expression was high compared the turbinate mucosa compared with normal controls, nasal polyps (P=0.01, P=0.01), Nod2protein expression showed no significant difference among the three groups.(P=0.14)4.Before and after desensitization therapy, Nod1expression was detected in peripheral blood mononuclear cells of patients with allergic rhinitis,before treatment (relative amount of1.06±0.13) compared with control group (0.29±0.05) and decreased expression (0.34±0.15,P=0.00) after treatment, the change of expression before and after the difference was statistically significant (P=0.00).5There was positive correlation between nasal IL-4, IL-6and Nod1Movement, r=0.751, p=0.001, r=0.826, p=0.000, closely related.Level of IL-4, IL-6, IL-10of mucosa in allergic rhinitis and deviated nasal septum group showed no statistically significant (Unit:pg/mg,15.22±1.17vs13.02±4.60,254.67±27.3vs 206.00±62.56,20.25±2.75vs24.11±3.81, P=0.53,0.27,0.20), but lower than in patients with nasal polyps group (25.80±2.20,1133.09±312.08,51.10±17.22, P=0.00,0.00,0.01). IFN-gamma was lower than a deviated nasal septum group (20.50±2.00vs28.19±7.63, P=0.04), while, compared with the nasal polyps group (19.89±5.01, P=0.86), there was no significant difference.6. Serum cytokine concentrations after Sublingual desensitization treatment showed:pre-treatment of IL-6(8.25±0.93) pg/ml,after treatment (6.39±0.71) pg/ml, P=0.00; IL-10(2.66±0.48) pg/ml, after treatment (4.78±0.44) pg/ml,(P=0.00); IFN-gamma (2.90±0.68) pg/ml, after treatment (9.82±0.85) pg/ml, increased (P=0.00). The IL-4of nasal secretions decreased (P=0.01), while IL-6, IL-10, IFN-gamma between two groups showed no significant difference (P=0.33, P=0.19, P=0.18).7after sublingual desensitization therapy, there was a negative correlation between Nod1expression change in peripheral blood mononuclear cells and IL-10change in peripheral blood of,r=-0.884, p=0.002, closely related. While,also, a positive correlation of IL-6, r=0.574, p=0.106. Conclusion1. Nod1, Nod2and NALP3expression was seen in the three groups of organizations, and the Nod1expression in allergic rhinitis group was lower than the control group and nasal polyps group, while,the NALP3was higher than the normal control group, nasal polyps group. It showed Nod1,NALP3may be involved in the pathogenesis of allergic rhinitis.2Nod1expression can be detected in peripheral blood mononuclear cells of allergic rhinitis before and after the sublingual desensitization treatmentAnd expression of Nod1of peripheral blood mononuclear cells reduced after treatment..Besides,the change of IL-10in the peripheral blood was negatively correlated with the change of Nod1. So,it seemed that Nod1may regulate IL-10changes and be involved in sublingual desensitization therapy.