Efeito genoprotetor in vitro de β-glucanas sobre linfócitos de frangos (Gallus gallus domesticus) expostos à aflatoxina B1

Detalhes bibliográficos
Autor(a) principal: Zimmermann, Carine Eloise Prestes
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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spelling 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
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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)
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instname_str Universidade Federal de Santa Maria (UFSM)
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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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