The Expressions Of Inflammatory Cytokines In The Tissue Of Chronic Rhinosinusitis And Their Significance

Chapter 1 Analysis of Inflammatory Cytokine Expressions in Chronic Rhinosinusitis with Antibody ChipObjective:To investigate the expressions of inflammatory cytokines in the nasal mucosa of the patients with chronic rhinosinusitis with or without nasal polyps,recurrent nasal polyps and in the normal nasal mucosa by using an antibody chip.Methods:The total proteins from the all patients' nasal membrane were labeled with biotin.The biotin-labeled proteins reacted with antibody chips on which the antibodies of 40 major inflammatory cytokines were prepared.The target proteins were conjugated with streptomycin antibody labeled by horseradish peroxidase (HRP),and signals were imaged by laser scanner.Results:(1) In comparison with normal control group,there were 15 cytokines whose expression enhanced more than 1.5 fold in the patients without NP;and there were 13 cytokines of more than 1.5 fold expressions in the patients with nasal polyps;and there were 9 cytokines of more than 1.5 fold expressions in the patients with RNP.The expressions of IL-11,MIP-1δ, MCP-2 and IL-6sR enhanced in all the three patient groups.(2) It is only eotaxin-2 whose expression enhanced in chronic rhinosinusitis with NP, and decreased in RNP and chronic rhinosinusitis without NP. Impressively,increasing expression of IL-8 occurred in patients with RNP,but did not in chronic rhinosinusitis with or without NP.(3) The expressions of cytokines including IL-1β,IL-12p40,IL-12p70,IL-17, Eotaxin and M-CSF enhanced in the chronic rhinosinusitis with or without NP,but not in RNP.It was showed that sTNFR-Ⅰand sTNFR-Ⅱup-regulated in RNP and chronic rhinosinusitis with NP,yet they were not increased in chronic rhinosinusitis without NP.Discussion and Conclusions:In our study,the differential expressions of a lot of cytokines in three types of inflammatory lesions of nasal mucosa were determined with a novel antibody array technology.It was found that the expressions of a large number of inflammatory cytokines enhanced in the above three types of nasal mucosa lesions,whereas the differential expressions were encountered commonly among the three groups.The result indicate that the three types of lesions may be developed by different mechanisms or by activation of different cell signal pathways.It provides clues and theory foundations for further study of molecular mechanism and biomrkers of the three types of inflammatory lesions.In addition,the significance of the cytokines including Eotaxin,sTNFR-Ⅰand sTNFR-Ⅱin the generation and development of nasal polyps need to be clarified.Also,the possible causative relationship between IL-8 and the recurrence of nasal polyps is worthy of further studing.We did not find similar reports by searching MEDLINE database on April 3rd,2009.Chapter 2 The Expression of IL-8 and MIP-1δProtein in Chronic Rhinosinusitis and its SignificanceObjective:To validate the differential expressions of IL-8 and MIP-1δprotein in patients with chronic rhinosinusitis with or without NP, recurrent nasalpolyps and normal nasal mucosa by semiquantitative Western-Blotting,and to observe their localization and expression in nasal mucosa,so as to preliminaryly select the biomarkers from the three above-mentioned groups with inflammatory affection of nasal mucosa so that the biomarkers could possibly be employed as indicators in the clinical classification and prognosis judgement of chronic rhinosinusitis. Methods:All samples were investigated by Western-Blotting stained with two step immunohistochemical methods to evaluate the expressions of IL-8 and MIP-1δ,respectively.Meanwhile,the correlation analysis between IL-8 and MIP-1δwere carried out.Results:(1) The expression of IL-8 in tissue of RNP is higher than that of chronic rhinosinusitis with or without NP(P＜0.05),also higher than that in normal nasal mucosa. But there is no significant difference of IL-8 expression among the groups of normal control and chronic rhinosinusitis with or without NP (P＞0.05).(2) As compared to normal controls,MIP-1δexpression increased in chronic rhinosinusitis with or without NP(P＜0.05),whereas, MIP-1δexpression in RNP was further higher than chronic rhinosinusitis with or without NP(P＜0.05).(3) The location of IL-8 and MIP-1δprotein were coincident in all samples,expressing in the cytoplasm and cellular membrane of nasal mucosa,glandular epithelium and inflammatory cells. The correlation coefficient betweeen IL-8 and MIP-1δwas 0.61. Discussion and Conclusions:(1) The location of IL-8 and MIP-1δprotein were coincident in all samples,suggesting that they possibly contribute to the same/similar cellular signal pathway or metabolism.(2) The higher expression of IL-8 and MIP-1δin the nasal mucosa of patients with recurrent nasal polyps indicate that these cytokines could possibly become clinical objective indicators for postoperative follow-up and prognosis judgement of nasal polyps,and also a possible clue in searching for the therapeutic targets for recurrent nasal polyps.