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      Efeito da mitomicina C em polipose nasossinusal eosinofílica, in vivo: dosagem de IL5 e GM-CSF, RT-PCR Translated title: Effect of mitomocin C in eosinophilic nasal polyposis, in vivo: concentration of IL5 and GM-CSF, RT-PCR

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          Abstract

          A polipose nasossinusal eosinofílica (PNS) é manifestação de uma doença inflamatória crônica na mucosa do nariz e nos seios paranasais caracterizada por infiltração de granulócitos eosinófilos. O fator responsável pela eosinofilia e manutenção dessas células com a perpetuação do processo inflamatório e formação polipóide é objeto constante de estudos. As citocinas como IL5 (interleucina 5) e GM-CSF (fator estimulador de colônia granulócito macrófago) aumentam a sobrevida dos eosinófilos e prolongam a sua presença no tecido polipóide, diminuindo o índice de apoptose eosinofílica. OBJETIVO: Avaliar o efeito da mitomicina C - MMC - por meio de aplicação tópica em pacientes portadores de PNS eosinofílica quanto à presença de IL5 e GM-CSF. CASUÍSTICA E MÉTODOS: Quinze pacientes portadores de PNS eosinofílica foram submetidos à aplicação tópica de MMC na concentração de 0,5mg/ml, 1ml, durante cinco minutos, na cavidade nasal direita, e submetidos à biópsia para RT-PCR 24hs após. O grupo-controle foi a cavidade nasal esquerda. O perfil de citocinas foi analisado para IL5 e GM-CSF. RESULTADOS: A comparação dos resultados de GM-CSF pré e pós-uso de MMC quando usamos o teste t pareado apresenta p=0,041. A comparação para IL5 resulta em p < 0,001. CONCLUSÃO: O uso de MMC em pacientes com PNS mostra redução com significância estatística par GM-CSF e importante significância para IL5.

          Translated abstract

          Eosinophilic nasosinusal polyposis is a chronic inflammatory infection with elevated infiltration of eosinophils, which presents high rate of recurrence after surgical treatment. The continuous inflammatory process that leads to the formation of polyps requires constant clinical treatment. Contributing to the maintenance of eosinophilia are cytokines IL5 (interleukin-5) and GM-CSF (granulocyte macrophages colony-stimulating factor), which show up in elevated concentrations. These oligoproteins diminish the rate of apoptosis and prolong the survival of eosinophils. AIM: By diminishing these cytokines, the action of Mitomycin C (MMC), an antineoplasic drug which inhibits the synthesis of DNA, was studied. In a recent study the power of this drug to cause apoptosis in eosinophils, in vitro, of nasal polyps was verified. METHODOLOGY: A biopsy of the nasal polyps was undertaken in 15 patients carriers of eosinophilic nasosinusal polyposis 24 hours after applying 0.5 mg/ml of MMC during five minutes. RT-PCR (reverse transcription of polymerase chain reaction) for IL5 and GM-CSF was the method used to obtain the results. RESULTS: The comparison of the results of GM-CSF pre- and post-application of MMC, when the paired T-test was used, showed p=0.041 and for IL5 we found p<0.001. CONCLUSION: Topic use of MMC in patients with eosinophilic nasosinusal polyposis shows statistically significant reduction for GM-CSF and significant and important reduction for IL5.

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          Most cited references27

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          IL-5 synthesis is upregulated in human nasal polyp tissue.

          In most nasal polyps, tissue eosinophilia is a striking finding, the pathologic mechanism of which is not understood. This study was performed to investigate a possibly distinct cytokine and chemokine pattern that could explain the characteristic tissue eosinophilia in nasal polyps. Polyps from 23 patients and turbinate tissue from 18 control subjects were investigated. The cytokine protein content (IL-1 beta, IL-3, IL-4, IL-5, IL-6, IL-8, IL-10, tumor necrosis factor-alpha, granulocyte-macrophage colony-stimulating factor, IL-1RA, RANTES, GRO-alpha) of tissue homogenates was measured by ELISA. Immunohistochemistry was performed in selected samples to detect IL-5+, major basic protein-positive, and EG2+ cells. IL-5 was detectable in only one sample of tissue from 18 control subjects but was found in 18 of 23 nasal polyps. Immunohistochemistry revealed an abundant number of IL-5+ cells, of which 69.5% could be identified as eosinophils by morphology. IL-6, IL-8, IL-10, tumor necrosis factor-alpha, GRO-alpha, and RANTES were detected in all specimens, without significant differences between groups (p > or = 0.05), whereas significnatly higher concentrations of IL-1 beta and IL-1RA were found in turbinate mucosa (p < or = 0.05). IL-3 was not detectable: granulocyte-macrophage colony-stimulating factor could only occasionally be found. This study indicates that IL-5 plays a key role in the pathophysiology of eosinophilic nasal polyps and may be produced by eosinophils.
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            Prolonged localized tissue effects from 5-minute exposures to fluorouracil and mitomycin C.

            Rabbits undergoing full-thickness glaucoma filtering surgery were exposed for 5 minutes to one of three intraoperative treatments: (1) distilled water; (2) fluorouracil, 50 mg/mL; or (3) mitomycin C, 0.4 mg/mL. Tissue samples were taken from the subconjunctival and scleral tissues at the treated area and 90 degrees and 180 degrees from the center of the treated area and the adjacent cornea 2 mm from the limbus, 1 hour, 5 days, and 30 days postoperatively. The biopsy specimens were then placed in tissue culture media and the fibroblast outgrowths measured. Five-minute intraoperative treatments with fluorouracil resulted in a reversible delay of fibroblast outgrowths from treated subconjunctival and scleral tissues of just over 1 week in this model, whereas treatment with mitomycin C, 0.4 mg/mL, resulted in prolonged inhibition of at least 30 days. These effects were localized to the area treated. The many clinical implications of these findings are discussed.
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              Enhanced soluble interleukin-5 receptor alpha expression in nasal polyposis.

              Alternative splicing of the interleukin-5 receptor alpha (IL-5Ralpha)-subunit leads to the generation of a signalling, membrane-anchored (TM) isoform, or a secreted [soluble (SOL)], antagonistic variant. Given the key role of IL-5 in eosinophil function, we investigated SOL IL-5Ralpha expression pattern in an eosinophil-associated disease such as nasal polyposis (NP). An SOL IL-5Ralpha enzyme-linked immunosorbent assay and quantitative real-time polymerase chain reaction (PCR) were established and applied in serum, nasal secretion and nasal tissue of controls (n = 12), and NP patients (n = 42) with or without asthma. Analysis of serum, nasal secretion, and nasal tissue samples revealed that SOL IL-5Ralpha protein concentrations were significantly increased in NP vs control tissue. Within the NP group, there was a significant up-regulation of SOL IL-5Ralpha in patients with systemic airway disease. These findings were confirmed at the mRNA level, using an optimized real-time reverse-transcriptase PCR procedure. This report demonstrates SOL IL-5Ralpha transcript and protein up-regulation in NP. Soluble IL-5Ralpha differentiates nasal polyps with or without concomitant asthma. As SOL IL-5Ralpha is strongly up-regulated for disease and has antagonistic properties in vitro, our studies shed new light on the mechanisms of specific immunomodulatory therapies, such as anti-IL-5.
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                Author and article information

                Journal
                rboto
                Revista Brasileira de Otorrinolaringologia
                Rev. Bras. Otorrinolaringol.
                ABORL-CCF Associação Brasileira de Otorrinolaringologia e Cirurgia Cérvico-Facial (São Paulo, SP, Brazil )
                0034-7299
                February 2006
                : 72
                : 1
                : 38-42
                Affiliations
                [05] orgnameUniversidade Federal de Minas Gerais orgdiv1departamento de parasitologia
                [08] orgnameEscola Paulista de Medicina
                [07] orgnameUniversidade Federal de Minas Gerais orgdiv1departamento de oftalmologia, otorrinolaringologia e fonoaudiologia
                [01] orgnameUFMG orgdiv1Faculdade de Medicina
                [06] orgnameUFMG
                [03] orgnameUFMG orgdiv1Hospital das Clínicas
                [02] orgnameCiências Médicas orgdiv1Faculdade de Medicina
                [04] orgnameSanta Casa de Misericórdia
                Article
                S0034-72992006000100007 S0034-7299(06)07200107
                10.1590/S0034-72992006000100007
                3703975a-d50f-4a40-84eb-9ad5b7a44bff

                This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.

                History
                : 23 September 2005
                : 23 September 2005
                Page count
                Figures: 0, Tables: 0, Equations: 0, References: 28, Pages: 5
                Product

                SciELO Brazil

                Categories
                Artigos Originais

                nasosinusal polyposis,eosinophils,mitomycin C,interleukin 5,GM-CSF,polipose nasossinusal,eosinófilos,mitomicina c,interleucina 5

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