Modelo experimental de conjuntivite alérgica: efeito do tratamento farmacológico com a proteína anti-inflamatória galectina - 1
Autor(a) principal: | |
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Data de Publicação: | 2014 |
Tipo de documento: | Dissertação |
Idioma: | por |
Título da fonte: | Repositório Institucional da UNESP |
Texto Completo: | http://hdl.handle.net/11449/122179 |
Resumo: | Allergic conjunctivitis (AC) is a complex disease of the immune system which includes mast cell degranulation in the conjunctiva and release of mediators that trigger inflammatory cell infiltration in this mucous membrane. Among the anti-inflammatory mediators, we highlight the protein galectin-1 (Gal-1) which regulates the process of leukocyte transmigration, release of cytokines and mast cell degranulation. However, its role in ocular inflammation has been little studied. BALB/c male mice were sensitized subcutaneously with 5μg of ovalbumin (OVA) on days 0 and 7 and challenged by eye drops containing OVA (250 μg) once daily (days 14, 15 and 16). For pharmacological treatment (days 14-16) animals received intraperitoneally recombinant Gal-1 (rGal-1; 0.3 μg /animal) or dexamethasone (Dex; 1 mg/kg). The clinical score was performed twenty minutes after OVA challenge (days14-16). After 4 and 24 hours of last OVA challenge, the animals were sacrificed and the followed analyses were performed: histopathology and quantification of inflammatory cells in the bulbar conjunctiva and blood; plasma anti-OVA IgE levels by ELISA; cytokines (IL-2, IL-4, IL-5, IL-10, IL-13, TNF-α and IFN-γ) and chemokines (RANTES, MCP- 1 and eotaxin) levels in the eyes, lacrimal fluid and lymph nodes by multiplex panel; and immunohistochemistry to detect Gal-1 expression in the conjunctiva. The CA was characterized by a significant increase in the score, especially on the 16th day (5.7 ± 0.3, P < 0.001)compared with controls (2.6 ± 0.6), followed by high anti-OVA IgE levels after 4 and 24h. Pharmacological treatments with rGal - 1(1.2 ± 0.5, P < 0.001) and Dex (3.0 ± 0.2, P < 0.05) decreased the clinical signs and rGal -1 reduced anti-IgE OVA levels(28.6 ± 6.3, P < 0.05), compared to the CA group (70 ± 12.8). Histopathological and quantitative analysis of inflammatory cellspresented a significant increase of mast cells,intravascular and transmigrated ... |
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Modelo experimental de conjuntivite alérgica: efeito do tratamento farmacológico com a proteína anti-inflamatória galectina - 1Biologia molecularProteínas - EstruturaGalectina 1Conjutivite alérgicaOlhos - InflamaçãoAgentes anti-inflamatoriosImuno-histoquímicaMolecular biologyAllergic conjunctivitis (AC) is a complex disease of the immune system which includes mast cell degranulation in the conjunctiva and release of mediators that trigger inflammatory cell infiltration in this mucous membrane. Among the anti-inflammatory mediators, we highlight the protein galectin-1 (Gal-1) which regulates the process of leukocyte transmigration, release of cytokines and mast cell degranulation. However, its role in ocular inflammation has been little studied. BALB/c male mice were sensitized subcutaneously with 5μg of ovalbumin (OVA) on days 0 and 7 and challenged by eye drops containing OVA (250 μg) once daily (days 14, 15 and 16). For pharmacological treatment (days 14-16) animals received intraperitoneally recombinant Gal-1 (rGal-1; 0.3 μg /animal) or dexamethasone (Dex; 1 mg/kg). The clinical score was performed twenty minutes after OVA challenge (days14-16). After 4 and 24 hours of last OVA challenge, the animals were sacrificed and the followed analyses were performed: histopathology and quantification of inflammatory cells in the bulbar conjunctiva and blood; plasma anti-OVA IgE levels by ELISA; cytokines (IL-2, IL-4, IL-5, IL-10, IL-13, TNF-α and IFN-γ) and chemokines (RANTES, MCP- 1 and eotaxin) levels in the eyes, lacrimal fluid and lymph nodes by multiplex panel; and immunohistochemistry to detect Gal-1 expression in the conjunctiva. The CA was characterized by a significant increase in the score, especially on the 16th day (5.7 ± 0.3, P < 0.001)compared with controls (2.6 ± 0.6), followed by high anti-OVA IgE levels after 4 and 24h. Pharmacological treatments with rGal - 1(1.2 ± 0.5, P < 0.001) and Dex (3.0 ± 0.2, P < 0.05) decreased the clinical signs and rGal -1 reduced anti-IgE OVA levels(28.6 ± 6.3, P < 0.05), compared to the CA group (70 ± 12.8). Histopathological and quantitative analysis of inflammatory cellspresented a significant increase of mast cells,intravascular and transmigrated ...A conjuntivite alérgica (CA) representa uma doença complexa do sistema imune envolvendo desgranulação dos mastócitos na conjuntiva e liberação de mediadores que provocam o influxo de células inflamatórias nessa membrana mucosa. Entre os mediadores anti-inflamatórios, destacamos a proteína galectina-1 (Gal-1) capaz de controlar o processo de transmigração dos leucócitos, liberação de citocinas e desgranulação de mastócitos. Contudo, seu papel nas inflamações oculares tem sido pouco estudado. Nos dias 0 e 7, camundongos machos Balb/c receberam a administração via subcutânea de ovalbumina (OVA; 5μg) e, nos dias 14, 15 e 16 foram desafiados com OVA (250μg) por instilação direta no saco conjuntival. Para os tratamentos farmacológicos (dias 14-16), foram administrados intraperitonealmente Gal-1 recombinante (rGal-1; 0,3 μg/animal) ou dexametasona (Dex; 1 mg/kg). O escore clínico foi realizado 20 minutos após o desafio. Após 4 e 24 horas do último desafio, os animais foram sacrificados e as análises realizadas por meio de: histopatologia e quantificação de células inflamatórias na conjuntiva bulbar e sangue; dosagem dos níveis de IgE anti OVA no plasma pela técnica de ELISA; dosagens das citocinas (IL-2, IL-4, IL-5, IL-10, IL-13, TNF-α e IFN-γ) e quimiocinas (RANTES, MCP-1 e eotaxina) nos olhos, fluido lacrimal e linfonodos, por meio de painel multiplex e, imuno-histoquímica para detectar a expressão da Gal-1 na conjuntiva. A CA foi caracterizada pelo aumento significante do escore, principaelmnte no 16º dia (5,7 ± 0,3; P < 0,001) comparado ao controle (2.6±0.6), seguido de nível elevado de IgE anti-OVA após 4 e 24h. Os tratamentos farmacológicos com rGal-1 (1,2 ± 0,5; P <0,001) e Dex (3,0 ± 0,2; P < 0,05) diminuíram os sinais clínicos, e rGal-1 reduziu o nível de IgE anti-OVA (28,6 ± 6,3; P < 0,05), comparado ao grupo CA (70 ± 12,8A). As análises histopatológicas e quantitativas de células ...Universidade Estadual Paulista (Unesp)Gil, Cristiane Damas [UNESP]Oliani, Sonia Maria [UNESP]Universidade Estadual Paulista (Unesp)Mello-Bochi, Cláudia Bosnic [UNESP]2015-04-09T12:28:27Z2015-04-09T12:28:27Z2014-02-25info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesis106 f. : il. color., tabs.application/pdfMELLO-BOCHI, Cláudia Bosnic. Modelo experimental de conjuntivite alérgica: efeito do tratamento farmacológico com a proteína anti-inflamatória galectina - 1. 2014. 106 f. Dissertação (mestrado) - Universidade Estadual Paulista Julio de Mesquita Filho, Instituto de Biociências, Letras e Ciências Exatas, 2014.http://hdl.handle.net/11449/122179000809772000809772.pdf33004153023P55102737730539655Alephreponame:Repositório Institucional da UNESPinstname:Universidade Estadual Paulista (UNESP)instacron:UNESPporinfo:eu-repo/semantics/openAccess2023-10-04T06:04:58Zoai:repositorio.unesp.br:11449/122179Repositório InstitucionalPUBhttp://repositorio.unesp.br/oai/requestopendoar:29462024-08-05T13:58:52.100719Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP)false |
dc.title.none.fl_str_mv |
Modelo experimental de conjuntivite alérgica: efeito do tratamento farmacológico com a proteína anti-inflamatória galectina - 1 |
title |
Modelo experimental de conjuntivite alérgica: efeito do tratamento farmacológico com a proteína anti-inflamatória galectina - 1 |
spellingShingle |
Modelo experimental de conjuntivite alérgica: efeito do tratamento farmacológico com a proteína anti-inflamatória galectina - 1 Mello-Bochi, Cláudia Bosnic [UNESP] Biologia molecular Proteínas - Estrutura Galectina 1 Conjutivite alérgica Olhos - Inflamação Agentes anti-inflamatorios Imuno-histoquímica Molecular biology |
title_short |
Modelo experimental de conjuntivite alérgica: efeito do tratamento farmacológico com a proteína anti-inflamatória galectina - 1 |
title_full |
Modelo experimental de conjuntivite alérgica: efeito do tratamento farmacológico com a proteína anti-inflamatória galectina - 1 |
title_fullStr |
Modelo experimental de conjuntivite alérgica: efeito do tratamento farmacológico com a proteína anti-inflamatória galectina - 1 |
title_full_unstemmed |
Modelo experimental de conjuntivite alérgica: efeito do tratamento farmacológico com a proteína anti-inflamatória galectina - 1 |
title_sort |
Modelo experimental de conjuntivite alérgica: efeito do tratamento farmacológico com a proteína anti-inflamatória galectina - 1 |
author |
Mello-Bochi, Cláudia Bosnic [UNESP] |
author_facet |
Mello-Bochi, Cláudia Bosnic [UNESP] |
author_role |
author |
dc.contributor.none.fl_str_mv |
Gil, Cristiane Damas [UNESP] Oliani, Sonia Maria [UNESP] Universidade Estadual Paulista (Unesp) |
dc.contributor.author.fl_str_mv |
Mello-Bochi, Cláudia Bosnic [UNESP] |
dc.subject.por.fl_str_mv |
Biologia molecular Proteínas - Estrutura Galectina 1 Conjutivite alérgica Olhos - Inflamação Agentes anti-inflamatorios Imuno-histoquímica Molecular biology |
topic |
Biologia molecular Proteínas - Estrutura Galectina 1 Conjutivite alérgica Olhos - Inflamação Agentes anti-inflamatorios Imuno-histoquímica Molecular biology |
description |
Allergic conjunctivitis (AC) is a complex disease of the immune system which includes mast cell degranulation in the conjunctiva and release of mediators that trigger inflammatory cell infiltration in this mucous membrane. Among the anti-inflammatory mediators, we highlight the protein galectin-1 (Gal-1) which regulates the process of leukocyte transmigration, release of cytokines and mast cell degranulation. However, its role in ocular inflammation has been little studied. BALB/c male mice were sensitized subcutaneously with 5μg of ovalbumin (OVA) on days 0 and 7 and challenged by eye drops containing OVA (250 μg) once daily (days 14, 15 and 16). For pharmacological treatment (days 14-16) animals received intraperitoneally recombinant Gal-1 (rGal-1; 0.3 μg /animal) or dexamethasone (Dex; 1 mg/kg). The clinical score was performed twenty minutes after OVA challenge (days14-16). After 4 and 24 hours of last OVA challenge, the animals were sacrificed and the followed analyses were performed: histopathology and quantification of inflammatory cells in the bulbar conjunctiva and blood; plasma anti-OVA IgE levels by ELISA; cytokines (IL-2, IL-4, IL-5, IL-10, IL-13, TNF-α and IFN-γ) and chemokines (RANTES, MCP- 1 and eotaxin) levels in the eyes, lacrimal fluid and lymph nodes by multiplex panel; and immunohistochemistry to detect Gal-1 expression in the conjunctiva. The CA was characterized by a significant increase in the score, especially on the 16th day (5.7 ± 0.3, P < 0.001)compared with controls (2.6 ± 0.6), followed by high anti-OVA IgE levels after 4 and 24h. Pharmacological treatments with rGal - 1(1.2 ± 0.5, P < 0.001) and Dex (3.0 ± 0.2, P < 0.05) decreased the clinical signs and rGal -1 reduced anti-IgE OVA levels(28.6 ± 6.3, P < 0.05), compared to the CA group (70 ± 12.8). Histopathological and quantitative analysis of inflammatory cellspresented a significant increase of mast cells,intravascular and transmigrated ... |
publishDate |
2014 |
dc.date.none.fl_str_mv |
2014-02-25 2015-04-09T12:28:27Z 2015-04-09T12:28:27Z |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/masterThesis |
format |
masterThesis |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
MELLO-BOCHI, Cláudia Bosnic. Modelo experimental de conjuntivite alérgica: efeito do tratamento farmacológico com a proteína anti-inflamatória galectina - 1. 2014. 106 f. Dissertação (mestrado) - Universidade Estadual Paulista Julio de Mesquita Filho, Instituto de Biociências, Letras e Ciências Exatas, 2014. http://hdl.handle.net/11449/122179 000809772 000809772.pdf 33004153023P5 5102737730539655 |
identifier_str_mv |
MELLO-BOCHI, Cláudia Bosnic. Modelo experimental de conjuntivite alérgica: efeito do tratamento farmacológico com a proteína anti-inflamatória galectina - 1. 2014. 106 f. Dissertação (mestrado) - Universidade Estadual Paulista Julio de Mesquita Filho, Instituto de Biociências, Letras e Ciências Exatas, 2014. 000809772 000809772.pdf 33004153023P5 5102737730539655 |
url |
http://hdl.handle.net/11449/122179 |
dc.language.iso.fl_str_mv |
por |
language |
por |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
eu_rights_str_mv |
openAccess |
dc.format.none.fl_str_mv |
106 f. : il. color., tabs. application/pdf |
dc.publisher.none.fl_str_mv |
Universidade Estadual Paulista (Unesp) |
publisher.none.fl_str_mv |
Universidade Estadual Paulista (Unesp) |
dc.source.none.fl_str_mv |
Aleph reponame:Repositório Institucional da UNESP instname:Universidade Estadual Paulista (UNESP) instacron:UNESP |
instname_str |
Universidade Estadual Paulista (UNESP) |
instacron_str |
UNESP |
institution |
UNESP |
reponame_str |
Repositório Institucional da UNESP |
collection |
Repositório Institucional da UNESP |
repository.name.fl_str_mv |
Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP) |
repository.mail.fl_str_mv |
|
_version_ |
1808128299534123008 |