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

Detalhes bibliográficos
Ano de defesa: 2013
Autor(a) principal: Zimmermann, Carine Eloise Prestes lattes
Orientador(a): Santurio, Janio Morais lattes
Banca de defesa: Leal, Daniela Bitencourt Rosa lattes, Loreto, érico Silva de lattes
Tipo de documento: Dissertação
Tipo de acesso: Acesso aberto
Idioma: por
Instituição de defesa: Universidade Federal de Santa Maria
Programa de Pós-Graduação: Programa de Pós-Graduação em Farmacologia
Departamento: Farmacologia
País: BR
Palavras-chave em Português:
Palavras-chave em Inglês:
Área do conhecimento CNPq:
Link de acesso: 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 2015-02-232015-02-232013-08-29ZIMMERMANN, 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/1/9003The 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.A 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.Coordenação de Aperfeiçoamento de Pessoal de Nível Superiorapplication/pdfporUniversidade Federal de Santa MariaPrograma de Pós-Graduação em FarmacologiaUFSMBRFarmacologiaAflatoxina B1β-glucanaLinfócitosFrangos de corteGenotoxicidadeAflatoxin B1β-glucansLymphocytesBroiler chickensGenotoxicityCNPQ::CIENCIAS BIOLOGICAS::FARMACOLOGIAEfeito 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 B1info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisSanturio, 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=K4713905Z8http://lattes.cnpq.br/2071516563093592Zimmermann, Carine Eloise Prestes20100000000040050030050050006a1b29c-fde0-4a69-8097-2a9d6a9d96494b17ea50-5e4b-4780-b44e-5ef9c71731342dbfa8b1-3511-410e-8203-4bb0cf639ac8ea251119-5531-432f-bf54-8c8111248727info:eu-repo/semantics/openAccessreponame:Biblioteca Digital de Teses e Dissertações do UFSMinstname:Universidade Federal de Santa Maria (UFSM)instacron:UFSMORIGINALZIMMERMANN, CARINE ELOISE PRESTES.pdfapplication/pdf826543http://repositorio.ufsm.br/bitstream/1/9003/1/ZIMMERMANN%2c%20CARINE%20ELOISE%20PRESTES.pdfa9347987db0d7ae894018616aeba7762MD51TEXTZIMMERMANN, CARINE ELOISE PRESTES.pdf.txtZIMMERMANN, CARINE ELOISE PRESTES.pdf.txtExtracted texttext/plain136403http://repositorio.ufsm.br/bitstream/1/9003/2/ZIMMERMANN%2c%20CARINE%20ELOISE%20PRESTES.pdf.txt60cd537209fc87cee0d73ccf3e602c11MD52THUMBNAILZIMMERMANN, CARINE ELOISE PRESTES.pdf.jpgZIMMERMANN, CARINE ELOISE PRESTES.pdf.jpgIM Thumbnailimage/jpeg5019http://repositorio.ufsm.br/bitstream/1/9003/3/ZIMMERMANN%2c%20CARINE%20ELOISE%20PRESTES.pdf.jpg5bb1f5b688e3e48ecccc2d4fdda4039dMD531/90032017-07-30 02:31:45.738oai: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:2017-07-30T05:31:45Biblioteca Digital de Teses e Dissertações do UFSM - Universidade Federal de Santa Maria (UFSM)false
dc.title.por.fl_str_mv Efeito genoprotetor in vitro de β-glucanas sobre linfócitos de frangos (Gallus gallus domesticus) expostos à aflatoxina B1
dc.title.alternative.eng.fl_str_mv 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.advisor1.fl_str_mv Santurio, Janio Morais
dc.contributor.advisor1Lattes.fl_str_mv http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4787502H6
dc.contributor.referee1.fl_str_mv Leal, Daniela Bitencourt Rosa
dc.contributor.referee1Lattes.fl_str_mv http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4706487E7
dc.contributor.referee2.fl_str_mv Loreto, érico Silva de
dc.contributor.referee2Lattes.fl_str_mv http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4713905Z8
dc.contributor.authorLattes.fl_str_mv http://lattes.cnpq.br/2071516563093592
dc.contributor.author.fl_str_mv Zimmermann, Carine Eloise Prestes
contributor_str_mv Santurio, Janio Morais
Leal, Daniela Bitencourt Rosa
Loreto, érico Silva de
dc.subject.por.fl_str_mv Aflatoxina B1
β-glucana
Linfócitos
Frangos de corte
Genotoxicidade
topic Aflatoxina B1
β-glucana
Linfócitos
Frangos de corte
Genotoxicidade
Aflatoxin B1
β-glucans
Lymphocytes
Broiler chickens
Genotoxicity
CNPQ::CIENCIAS BIOLOGICAS::FARMACOLOGIA
dc.subject.eng.fl_str_mv Aflatoxin B1
β-glucans
Lymphocytes
Broiler chickens
Genotoxicity
dc.subject.cnpq.fl_str_mv 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.issued.fl_str_mv 2013-08-29
dc.date.accessioned.fl_str_mv 2015-02-23
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dc.identifier.citation.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.
dc.identifier.uri.fl_str_mv 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
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