Efeito genoprotetor in vitro de β-glucanas sobre linfócitos de frangos (Gallus gallus domesticus) expostos à aflatoxina B1
Autor(a) principal: | |
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Data de Publicação: | 2013 |
Tipo de documento: | Dissertação |
Idioma: | por |
Título da fonte: | Manancial - Repositório Digital da UFSM |
Texto Completo: | http://repositorio.ufsm.br/handle/1/9003 |
Resumo: | The aflatoxin B1 (AFB1) is one of the main mycotoxins that can be identified in foods, and has relevance for agricultural economics and public health because of its immunotoxic properties. A functional immune system is a basic requirement for a healthy life in modern animal production. The interaction involving nutrition and immunity is a strategic factor to obtain a high quality performance in the poultry industry. Immunomodulators such as β-glucans have an immunostimulating activity, which enables the host ability to resist opportunistic infections. To contribute to the elucidation of the mechanism of action of β-glucans in broiler chicken lymphocytes, the effects of the concentrations of 0.1, 1 and 10% of β-glucans derived from Saccharomyces cerevisiae were investigated in lymphocytes exposed to increasing concentrations of AFB1 (0, 0.1, 1, 10, 20 μg/ml). Lymphocytes were separated by Ficoll-Histopaque density and cultured in 96 well-plates containing AFB1 and/or β-glucans in a 5% CO2 atmosphere at 39°C. MTT, PicoGreen® and 2'-7'-dichlorofluorescein diacetate cytotoxicity tests were evaluated at 24, 48 and 72 h of incubation. The comet assay to elucidate the DNA damage was also performed. The percentage of viable cells decreased in the presence of 10 μg/ml AFB1 at 48 h (p < 0.05) and 10 and 20 μg/ml AFB1 at 72 h (p < 0.001 and p < 0.01, respectively), when compared to the control group (0 μg/ml). Furthermore, an increase in cell-free DNA in AFB1 concentrations > 1 μg/ml (p < 0.001) and the generation of ROS at 24 h were also observed. DNA damage increased approximately 2.3 fold in lymphocytes exposed to 20 μg/ml of AFB1 when compared to the control group. Conversely, β-glucans showed cytoprotective effects (p < 0.001), and the concentration of 1% reverted the AFB1-induced lymphocyte damage. β-glucans at 10% significantly increased (p < 0.001) the formation of reactive oxygen species (ROS), potentiating the AFB1-induced ROS formation. In conclusion, this study showed that AFB1 and β-glucans exert influence on lymphocyte oxidative metabolism and have dose-dependent potentiating effects. The results also showed genoprotective in vitro effect of β-glucans in poultry lymphocytes exposed to AFB1, being the concentration of 1% β-glucans able to maintain DNA integrity. |
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Efeito genoprotetor in vitro de β-glucanas sobre linfócitos de frangos (Gallus gallus domesticus) expostos à aflatoxina B1In vitro genoprotective effect of β-glucan on broiler chicken (Gallus gallus domesticus) lymphocytes exposed to aflatoxin B1Aflatoxina B1β-glucanaLinfócitosFrangos de corteGenotoxicidadeAflatoxin B1β-glucansLymphocytesBroiler chickensGenotoxicityCNPQ::CIENCIAS BIOLOGICAS::FARMACOLOGIAThe aflatoxin B1 (AFB1) is one of the main mycotoxins that can be identified in foods, and has relevance for agricultural economics and public health because of its immunotoxic properties. A functional immune system is a basic requirement for a healthy life in modern animal production. The interaction involving nutrition and immunity is a strategic factor to obtain a high quality performance in the poultry industry. Immunomodulators such as β-glucans have an immunostimulating activity, which enables the host ability to resist opportunistic infections. To contribute to the elucidation of the mechanism of action of β-glucans in broiler chicken lymphocytes, the effects of the concentrations of 0.1, 1 and 10% of β-glucans derived from Saccharomyces cerevisiae were investigated in lymphocytes exposed to increasing concentrations of AFB1 (0, 0.1, 1, 10, 20 μg/ml). Lymphocytes were separated by Ficoll-Histopaque density and cultured in 96 well-plates containing AFB1 and/or β-glucans in a 5% CO2 atmosphere at 39°C. MTT, PicoGreen® and 2'-7'-dichlorofluorescein diacetate cytotoxicity tests were evaluated at 24, 48 and 72 h of incubation. The comet assay to elucidate the DNA damage was also performed. The percentage of viable cells decreased in the presence of 10 μg/ml AFB1 at 48 h (p < 0.05) and 10 and 20 μg/ml AFB1 at 72 h (p < 0.001 and p < 0.01, respectively), when compared to the control group (0 μg/ml). Furthermore, an increase in cell-free DNA in AFB1 concentrations > 1 μg/ml (p < 0.001) and the generation of ROS at 24 h were also observed. DNA damage increased approximately 2.3 fold in lymphocytes exposed to 20 μg/ml of AFB1 when compared to the control group. Conversely, β-glucans showed cytoprotective effects (p < 0.001), and the concentration of 1% reverted the AFB1-induced lymphocyte damage. β-glucans at 10% significantly increased (p < 0.001) the formation of reactive oxygen species (ROS), potentiating the AFB1-induced ROS formation. In conclusion, this study showed that AFB1 and β-glucans exert influence on lymphocyte oxidative metabolism and have dose-dependent potentiating effects. The results also showed genoprotective in vitro effect of β-glucans in poultry lymphocytes exposed to AFB1, being the concentration of 1% β-glucans able to maintain DNA integrity.Coordenação de Aperfeiçoamento de Pessoal de Nível SuperiorA aflatoxina B1 (AFB1) é uma das principais micotoxinas que podem ser identificadas em alimentos, possuindo relevância agroeconômica e para a saúde pública, por ser considerada imunotóxica. Um sistema imune funcional é um requisito básico para uma vida saudável na produção moderna de animais. A interação envolvendo nutrição e imunidade é um fator estratégico para obter um bom desempenho em frangos de corte. Devido as suas atividades imunomodulatórias, substâncias como as β-glucanas proporcionam ao hospedeiro uma maior capacidade de resistir a infecções oportunistas. Para melhor compreender o mecanismo de ação das β-glucanas, sobre os linfócitos de frangos de corte, investigou-se os efeitos das concentrações de 0.1, 1 e 10% de β-glucanas derivadas do fungo Saccharomyces cerevisiae em linfócitos expostos a crescentes concentrações de AFB1 (0, 0.1, 1, 10, 20 μg/ml). Os linfócitos foram separados através do reagente de densidade Ficoll-Histopaque e cultivados em placas de 96 poços, contendo as concentrações de AFB1 e/ou β-glucanas em atmosfera de 5% de CO2 a 39°C durante 24, 48 e 72 h. A citotoxicidade celular foi avaliada através dos testes MTT e PicoGreen®, e a formação de espécies reativas de oxigênio (EROS) através do ensaio 2′-7′- diacetato diclorofluoresceína. Também foi utilizado o teste cometa para elucidar os danos ao DNA. A viabilidade celular reduziu na presença de 10 μg/ml de AFB1 em 48 h (p < 0.05) e em 10 and 20 μg/ml de AFB1 (p < 0.01 e p < 0.001, respectivamente) em 72 h quando comparada ao grupo controle. Além disso, as concentrações de AFB1 > 1 μg/ml aumentaram significativamente (p < 0.001) a liberação de fita dupla de DNA (dsDNA) e a produção de EROS em 24 h. Os danos causados ao DNA foram confirmados através do momento cauda do cometa e aumentaram cerca de 2,3 vezes em linfócitos expostos a 20 μg/ml de AFB1 quando comparados ao grupo controle. Por outro lado, as β-glucanas exerceram efeitos citoprotetores (p < 0.001), sendo que a concentração de 1% foi capaz de reverter os danos genotóxicos causados pela ação da AFB1. Já a concentração de 10% de β-glucanas aumentou significativamente (p < 0.001) a formação de EROS, potencializando a ação da AFB1. Em conclusão, este estudou evidenciou que a AFB1 e as β-glucanas exercem influência sobre o metabolismo oxidativo dos linfócitos e possui efeito potencializador dose-dependente. Os resultados também evidenciaram o efeito genoprotetor in vitro de β-glucanas em linfócitos de frangos expostos a AFB1, sendo a concentração de β-glucanas a 1% capaz de manter a integridade do DNA.Universidade Federal de Santa MariaBRFarmacologiaUFSMPrograma de Pós-Graduação em FarmacologiaSanturio, Janio Moraishttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4787502H6Leal, Daniela Bitencourt Rosahttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4706487E7Loreto, érico Silva dehttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4713905Z8Zimmermann, Carine Eloise Prestes2015-02-232015-02-232013-08-29info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisapplication/pdfapplication/pdfZIMMERMANN, Carine Eloise Prestes. IN VITRO GENOPROTECTIVE EFFECT OF β-GLUCANS ON BROILER CHICKEN (Gallus gallus domesticus) LYMPHOCYTES EXPOSED TO AFLATOXIN B1. 2013. 79 f. Dissertação (Mestrado em Farmácia) - Universidade Federal de Santa Maria, Santa Maria, 2013.http://repositorio.ufsm.br/handle/1/9003porinfo:eu-repo/semantics/openAccessreponame:Manancial - Repositório Digital da UFSMinstname:Universidade Federal de Santa Maria (UFSM)instacron:UFSM2017-07-30T05:31:45Zoai:repositorio.ufsm.br:1/9003Biblioteca Digital de Teses e Dissertaçõeshttps://repositorio.ufsm.br/ONGhttps://repositorio.ufsm.br/oai/requestatendimento.sib@ufsm.br||tedebc@gmail.comopendoar:2017-07-30T05:31:45Manancial - Repositório Digital da UFSM - Universidade Federal de Santa Maria (UFSM)false |
dc.title.none.fl_str_mv |
Efeito genoprotetor in vitro de β-glucanas sobre linfócitos de frangos (Gallus gallus domesticus) expostos à aflatoxina B1 In vitro genoprotective effect of β-glucan on broiler chicken (Gallus gallus domesticus) lymphocytes exposed to aflatoxin B1 |
title |
Efeito genoprotetor in vitro de β-glucanas sobre linfócitos de frangos (Gallus gallus domesticus) expostos à aflatoxina B1 |
spellingShingle |
Efeito genoprotetor in vitro de β-glucanas sobre linfócitos de frangos (Gallus gallus domesticus) expostos à aflatoxina B1 Zimmermann, Carine Eloise Prestes Aflatoxina B1 β-glucana Linfócitos Frangos de corte Genotoxicidade Aflatoxin B1 β-glucans Lymphocytes Broiler chickens Genotoxicity CNPQ::CIENCIAS BIOLOGICAS::FARMACOLOGIA |
title_short |
Efeito genoprotetor in vitro de β-glucanas sobre linfócitos de frangos (Gallus gallus domesticus) expostos à aflatoxina B1 |
title_full |
Efeito genoprotetor in vitro de β-glucanas sobre linfócitos de frangos (Gallus gallus domesticus) expostos à aflatoxina B1 |
title_fullStr |
Efeito genoprotetor in vitro de β-glucanas sobre linfócitos de frangos (Gallus gallus domesticus) expostos à aflatoxina B1 |
title_full_unstemmed |
Efeito genoprotetor in vitro de β-glucanas sobre linfócitos de frangos (Gallus gallus domesticus) expostos à aflatoxina B1 |
title_sort |
Efeito genoprotetor in vitro de β-glucanas sobre linfócitos de frangos (Gallus gallus domesticus) expostos à aflatoxina B1 |
author |
Zimmermann, Carine Eloise Prestes |
author_facet |
Zimmermann, Carine Eloise Prestes |
author_role |
author |
dc.contributor.none.fl_str_mv |
Santurio, Janio Morais http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4787502H6 Leal, Daniela Bitencourt Rosa http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4706487E7 Loreto, érico Silva de http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4713905Z8 |
dc.contributor.author.fl_str_mv |
Zimmermann, Carine Eloise Prestes |
dc.subject.por.fl_str_mv |
Aflatoxina B1 β-glucana Linfócitos Frangos de corte Genotoxicidade Aflatoxin B1 β-glucans Lymphocytes Broiler chickens Genotoxicity CNPQ::CIENCIAS BIOLOGICAS::FARMACOLOGIA |
topic |
Aflatoxina B1 β-glucana Linfócitos Frangos de corte Genotoxicidade Aflatoxin B1 β-glucans Lymphocytes Broiler chickens Genotoxicity CNPQ::CIENCIAS BIOLOGICAS::FARMACOLOGIA |
description |
The aflatoxin B1 (AFB1) is one of the main mycotoxins that can be identified in foods, and has relevance for agricultural economics and public health because of its immunotoxic properties. A functional immune system is a basic requirement for a healthy life in modern animal production. The interaction involving nutrition and immunity is a strategic factor to obtain a high quality performance in the poultry industry. Immunomodulators such as β-glucans have an immunostimulating activity, which enables the host ability to resist opportunistic infections. To contribute to the elucidation of the mechanism of action of β-glucans in broiler chicken lymphocytes, the effects of the concentrations of 0.1, 1 and 10% of β-glucans derived from Saccharomyces cerevisiae were investigated in lymphocytes exposed to increasing concentrations of AFB1 (0, 0.1, 1, 10, 20 μg/ml). Lymphocytes were separated by Ficoll-Histopaque density and cultured in 96 well-plates containing AFB1 and/or β-glucans in a 5% CO2 atmosphere at 39°C. MTT, PicoGreen® and 2'-7'-dichlorofluorescein diacetate cytotoxicity tests were evaluated at 24, 48 and 72 h of incubation. The comet assay to elucidate the DNA damage was also performed. The percentage of viable cells decreased in the presence of 10 μg/ml AFB1 at 48 h (p < 0.05) and 10 and 20 μg/ml AFB1 at 72 h (p < 0.001 and p < 0.01, respectively), when compared to the control group (0 μg/ml). Furthermore, an increase in cell-free DNA in AFB1 concentrations > 1 μg/ml (p < 0.001) and the generation of ROS at 24 h were also observed. DNA damage increased approximately 2.3 fold in lymphocytes exposed to 20 μg/ml of AFB1 when compared to the control group. Conversely, β-glucans showed cytoprotective effects (p < 0.001), and the concentration of 1% reverted the AFB1-induced lymphocyte damage. β-glucans at 10% significantly increased (p < 0.001) the formation of reactive oxygen species (ROS), potentiating the AFB1-induced ROS formation. In conclusion, this study showed that AFB1 and β-glucans exert influence on lymphocyte oxidative metabolism and have dose-dependent potentiating effects. The results also showed genoprotective in vitro effect of β-glucans in poultry lymphocytes exposed to AFB1, being the concentration of 1% β-glucans able to maintain DNA integrity. |
publishDate |
2013 |
dc.date.none.fl_str_mv |
2013-08-29 2015-02-23 2015-02-23 |
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 |
ZIMMERMANN, Carine Eloise Prestes. IN VITRO GENOPROTECTIVE EFFECT OF β-GLUCANS ON BROILER CHICKEN (Gallus gallus domesticus) LYMPHOCYTES EXPOSED TO AFLATOXIN B1. 2013. 79 f. Dissertação (Mestrado em Farmácia) - Universidade Federal de Santa Maria, Santa Maria, 2013. http://repositorio.ufsm.br/handle/1/9003 |
identifier_str_mv |
ZIMMERMANN, Carine Eloise Prestes. IN VITRO GENOPROTECTIVE EFFECT OF β-GLUCANS ON BROILER CHICKEN (Gallus gallus domesticus) LYMPHOCYTES EXPOSED TO AFLATOXIN B1. 2013. 79 f. Dissertação (Mestrado em Farmácia) - Universidade Federal de Santa Maria, Santa Maria, 2013. |
url |
http://repositorio.ufsm.br/handle/1/9003 |
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 |
application/pdf application/pdf |
dc.publisher.none.fl_str_mv |
Universidade Federal de Santa Maria BR Farmacologia UFSM Programa de Pós-Graduação em Farmacologia |
publisher.none.fl_str_mv |
Universidade Federal de Santa Maria BR Farmacologia UFSM Programa de Pós-Graduação em Farmacologia |
dc.source.none.fl_str_mv |
reponame:Manancial - Repositório Digital da UFSM instname:Universidade Federal de Santa Maria (UFSM) instacron:UFSM |
instname_str |
Universidade Federal de Santa Maria (UFSM) |
instacron_str |
UFSM |
institution |
UFSM |
reponame_str |
Manancial - Repositório Digital da UFSM |
collection |
Manancial - Repositório Digital da UFSM |
repository.name.fl_str_mv |
Manancial - Repositório Digital da UFSM - Universidade Federal de Santa Maria (UFSM) |
repository.mail.fl_str_mv |
atendimento.sib@ufsm.br||tedebc@gmail.com |
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1805922175737135104 |