Avaliação dos efeitos do eugenol e do óleo essencial de Ocimum gratissimum durante o cultivo in vitro de folículos pré-antrais bovinos

Detalhes bibliográficos
Autor(a) principal: Vasconcelos, Erlândia Márcia
Data de Publicação: 2020
Tipo de documento: Dissertação
Idioma: por
Título da fonte: Repositório Institucional da Universidade Federal do Ceará (UFC)
Texto Completo: http://www.repositorio.ufc.br/handle/riufc/53680
Resumo: The aim of this work was to evaluate the effects of different concentrations of Eugenol and the essential oil of Ocimum gratissimum on growth, viability, morphology, antrum formation and mRNA expression for SOD, CAT, GPX1, and PRDX6 in secondary bovine follicles grown in vitro. Bovine ovaries were collected in a local slaughterhouse, under aseptic conditions. In the laboratory, the secondary follicles were isolated from the ovarian cortex and cultured in TCM- 199 + drops alone or supplemented with different concentrations of the essential oil of Ocimum gratissimum (0.125%, 0.25% and 0.5%) and Eugenol (5, 10 and 50 μM). The culture medium was changed every 2 days and the photographic record of the follicles was done on days 0, 6, 12 and 18, with growth rate and antrum formation being observed. At the end of the cultivation period, the follicles were subjected to viability analysis using calcein (viable) and homodimer ethidium (non-viable). In addition, follicles were used to quantify messenger RNA levels for SOD, CAT, GPX1 and PRDX6. The data on follicular growth were submitted to ANOVA, followed by the T test. For the analysis of follicular viability and antrum formation, the Chisquare test was used (p <0.05). The messenger RNA expression data were analyzed using the nonparametric Kruskal-Wallis test (p <0.05). The results showed that the secondary follicles grown in the presence of the essential oil of O. gratissiumum maintained morphology, viability and growth. In addition, the presence of 0.5% or 0.25% essential oil in the culture medium significantly increased the levels of mRNAs for PRDX6 and CAT. Secondary follicles grown with Eugenol maintained similar morphology and viability to the control group. A progressive increase in follicular diameter was observed between days 0 and 12 in all treatments, except for follicles grown with 50 μM Eugenol. 5.0 and 50.0 μΜ Eugenol increased mRNA levels for PRDX6 in follicles, but 0.5 μΜ Eugenol reduced mRNA levels for SOD in cultured follicles. Eugenol did not influence mRNA expression for CAT and GPX1. In conclusion, O. gratissimum essential oil increases the levels of mRNAs for PRDX6 and CAT in secondary bovine follicles grown in vitro for 18 days and maintains their viability, morphology and growth. Eugenol reduces the levels of mRNAs for SOD and increases the levels of PRDX6 in bovine secondary follicles grown in vitro. Eugenol maintains follicular viability and morphology, but reduces follicular growth when in high concentration (50 μM).
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spelling Avaliação dos efeitos do eugenol e do óleo essencial de Ocimum gratissimum durante o cultivo in vitro de folículos pré-antrais bovinosEvaluation of the effects of eugenol and the essential oil of Ocimum gratissimum during in vitro cultivation of bovine preantral folliclesFolículosEstresse oxidativoAntioxidanteThe aim of this work was to evaluate the effects of different concentrations of Eugenol and the essential oil of Ocimum gratissimum on growth, viability, morphology, antrum formation and mRNA expression for SOD, CAT, GPX1, and PRDX6 in secondary bovine follicles grown in vitro. Bovine ovaries were collected in a local slaughterhouse, under aseptic conditions. In the laboratory, the secondary follicles were isolated from the ovarian cortex and cultured in TCM- 199 + drops alone or supplemented with different concentrations of the essential oil of Ocimum gratissimum (0.125%, 0.25% and 0.5%) and Eugenol (5, 10 and 50 μM). The culture medium was changed every 2 days and the photographic record of the follicles was done on days 0, 6, 12 and 18, with growth rate and antrum formation being observed. At the end of the cultivation period, the follicles were subjected to viability analysis using calcein (viable) and homodimer ethidium (non-viable). In addition, follicles were used to quantify messenger RNA levels for SOD, CAT, GPX1 and PRDX6. The data on follicular growth were submitted to ANOVA, followed by the T test. For the analysis of follicular viability and antrum formation, the Chisquare test was used (p <0.05). The messenger RNA expression data were analyzed using the nonparametric Kruskal-Wallis test (p <0.05). The results showed that the secondary follicles grown in the presence of the essential oil of O. gratissiumum maintained morphology, viability and growth. In addition, the presence of 0.5% or 0.25% essential oil in the culture medium significantly increased the levels of mRNAs for PRDX6 and CAT. Secondary follicles grown with Eugenol maintained similar morphology and viability to the control group. A progressive increase in follicular diameter was observed between days 0 and 12 in all treatments, except for follicles grown with 50 μM Eugenol. 5.0 and 50.0 μΜ Eugenol increased mRNA levels for PRDX6 in follicles, but 0.5 μΜ Eugenol reduced mRNA levels for SOD in cultured follicles. Eugenol did not influence mRNA expression for CAT and GPX1. In conclusion, O. gratissimum essential oil increases the levels of mRNAs for PRDX6 and CAT in secondary bovine follicles grown in vitro for 18 days and maintains their viability, morphology and growth. Eugenol reduces the levels of mRNAs for SOD and increases the levels of PRDX6 in bovine secondary follicles grown in vitro. Eugenol maintains follicular viability and morphology, but reduces follicular growth when in high concentration (50 μM).O objetivo deste trabalho foi avaliar os efeitos de diferentes concentrações do Eugenol e do óleo essencial de Ocimum gratissimum sobre o crescimento, viabilidade, morfologia, formação de antro e expressão de mRNA para SOD, CAT, GPX1, e PRDX6 em folículos secundários bovinos cultivados in vitro. Os ovários bovinos foram coletados em abatedouro local, em condições assépticas. No laboratório, os folículos foram isolados do córtex ovariano e cultivados em gotas de TCM-199+ sozinho ou suplementado com diferentes concentrações do óleo essencial de O. gratissimum (0,125%, 0,25% e 0,5%) e Eugenol (0,5, 5 e 50 μM). A troca do meio de cultivo foi feita a cada 2 dias e o registro fotográfico dos folículos foi feito nos dias 0, 6, 12 e 18, sendo observado a taxa de crescimento e formação de antro. Ao final do cultivo, os folículos foram submetidos à análise de viabilidade utilizando calceína e etídio homodímero. Os folículos foram utilizados para quantificar os níveis de RNA mensageiros para SOD, CAT, GPX1 e PRDX6. Os dados referentes ao crescimento folicular foram submetidos a ANOVA, seguido do teste T. Para a análise da viabilidade folicular e formação de antro foi utilizado o teste Qui-quadrado (p< 0,05). Os dados de expressão de RNA mensageiros foram analisados utilizando o teste de Kruskal-Wallis não paramétrico (p < 0,05). Os resultados mostraram que os folículos secundários cultivados na presença do óleo essencial de O. gratissiumum mantiveram morfologia, viabilidade e crescimento. A presença de 0,5% ou 0,25% de óleo essencial no meio de cultura aumentou significativamente os níveis de mRNAs para PRDX6 e CAT. Folículos secundários cultivados com Eugenol mantiveram morfologia e viabilidade semelhantes ao grupo controle. Um aumento progressivo no diâmetro folicular foi observado entre os dias 0 e 12 em todos os tratamentos, exceto nos folículos cultivados com Eugenol 50 μM. 5,0 e 50 μΜ Eugenol aumentaram os níveis de mRNA para PRDX6 nos folículos, mas 0,5 μΜ Eugenol reduziu os níveis de mRNA para SOD nos folículos cultivados. Eugenol não influenciou a expressão de mRNA para CAT e GPX1. Em conclusão, o óleo essencial de O. gratissimum aumenta os níveis de mRNAs para PRDX6 e CAT em folículos secundários bovinos cultivados por 18 dias e mantém viabilidade, morfologia e crescimento. Eugenol reduz níveis de mRNAs para SOD e aumenta os níveis de PRDX6 em folículos secundários bovinos os cultivados in vitro. O eugenol mantem a viabilidade e morfologia foliculares, mas reduz o crescimento folicular quando em alta concentração (50 μM).Silva, José Roberto VianaVasconcelos, Erlândia Márcia2020-08-27T11:43:30Z2020-08-27T11:43:30Z2020-02-07info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisapplication/pdfVasconcelos.E.M. Avaliação dos efeitos do eugenol e do óleo essencial de Ocimum gratissimum durante o cultivo in vitro de folículos pré-antrais bovinos. 2020. 98 f. Dissertação (Mestrado em Biotecnologia), Programa de Pós-Graduação em Biotecnologia - Campus de Sobral, Universidade Federal do Ceará, Sobral, 2020.http://www.repositorio.ufc.br/handle/riufc/53680porreponame:Repositório Institucional da Universidade Federal do Ceará (UFC)instname:Universidade Federal do Ceará (UFC)instacron:UFCinfo:eu-repo/semantics/openAccess2020-08-27T11:43:30Zoai:repositorio.ufc.br:riufc/53680Repositório InstitucionalPUBhttp://www.repositorio.ufc.br/ri-oai/requestbu@ufc.br || repositorio@ufc.bropendoar:2020-08-27T11:43:30Repositório Institucional da Universidade Federal do Ceará (UFC) - Universidade Federal do Ceará (UFC)false
dc.title.none.fl_str_mv Avaliação dos efeitos do eugenol e do óleo essencial de Ocimum gratissimum durante o cultivo in vitro de folículos pré-antrais bovinos
Evaluation of the effects of eugenol and the essential oil of Ocimum gratissimum during in vitro cultivation of bovine preantral follicles
title Avaliação dos efeitos do eugenol e do óleo essencial de Ocimum gratissimum durante o cultivo in vitro de folículos pré-antrais bovinos
spellingShingle Avaliação dos efeitos do eugenol e do óleo essencial de Ocimum gratissimum durante o cultivo in vitro de folículos pré-antrais bovinos
Vasconcelos, Erlândia Márcia
Folículos
Estresse oxidativo
Antioxidante
title_short Avaliação dos efeitos do eugenol e do óleo essencial de Ocimum gratissimum durante o cultivo in vitro de folículos pré-antrais bovinos
title_full Avaliação dos efeitos do eugenol e do óleo essencial de Ocimum gratissimum durante o cultivo in vitro de folículos pré-antrais bovinos
title_fullStr Avaliação dos efeitos do eugenol e do óleo essencial de Ocimum gratissimum durante o cultivo in vitro de folículos pré-antrais bovinos
title_full_unstemmed Avaliação dos efeitos do eugenol e do óleo essencial de Ocimum gratissimum durante o cultivo in vitro de folículos pré-antrais bovinos
title_sort Avaliação dos efeitos do eugenol e do óleo essencial de Ocimum gratissimum durante o cultivo in vitro de folículos pré-antrais bovinos
author Vasconcelos, Erlândia Márcia
author_facet Vasconcelos, Erlândia Márcia
author_role author
dc.contributor.none.fl_str_mv Silva, José Roberto Viana
dc.contributor.author.fl_str_mv Vasconcelos, Erlândia Márcia
dc.subject.por.fl_str_mv Folículos
Estresse oxidativo
Antioxidante
topic Folículos
Estresse oxidativo
Antioxidante
description The aim of this work was to evaluate the effects of different concentrations of Eugenol and the essential oil of Ocimum gratissimum on growth, viability, morphology, antrum formation and mRNA expression for SOD, CAT, GPX1, and PRDX6 in secondary bovine follicles grown in vitro. Bovine ovaries were collected in a local slaughterhouse, under aseptic conditions. In the laboratory, the secondary follicles were isolated from the ovarian cortex and cultured in TCM- 199 + drops alone or supplemented with different concentrations of the essential oil of Ocimum gratissimum (0.125%, 0.25% and 0.5%) and Eugenol (5, 10 and 50 μM). The culture medium was changed every 2 days and the photographic record of the follicles was done on days 0, 6, 12 and 18, with growth rate and antrum formation being observed. At the end of the cultivation period, the follicles were subjected to viability analysis using calcein (viable) and homodimer ethidium (non-viable). In addition, follicles were used to quantify messenger RNA levels for SOD, CAT, GPX1 and PRDX6. The data on follicular growth were submitted to ANOVA, followed by the T test. For the analysis of follicular viability and antrum formation, the Chisquare test was used (p <0.05). The messenger RNA expression data were analyzed using the nonparametric Kruskal-Wallis test (p <0.05). The results showed that the secondary follicles grown in the presence of the essential oil of O. gratissiumum maintained morphology, viability and growth. In addition, the presence of 0.5% or 0.25% essential oil in the culture medium significantly increased the levels of mRNAs for PRDX6 and CAT. Secondary follicles grown with Eugenol maintained similar morphology and viability to the control group. A progressive increase in follicular diameter was observed between days 0 and 12 in all treatments, except for follicles grown with 50 μM Eugenol. 5.0 and 50.0 μΜ Eugenol increased mRNA levels for PRDX6 in follicles, but 0.5 μΜ Eugenol reduced mRNA levels for SOD in cultured follicles. Eugenol did not influence mRNA expression for CAT and GPX1. In conclusion, O. gratissimum essential oil increases the levels of mRNAs for PRDX6 and CAT in secondary bovine follicles grown in vitro for 18 days and maintains their viability, morphology and growth. Eugenol reduces the levels of mRNAs for SOD and increases the levels of PRDX6 in bovine secondary follicles grown in vitro. Eugenol maintains follicular viability and morphology, but reduces follicular growth when in high concentration (50 μM).
publishDate 2020
dc.date.none.fl_str_mv 2020-08-27T11:43:30Z
2020-08-27T11:43:30Z
2020-02-07
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 Vasconcelos.E.M. Avaliação dos efeitos do eugenol e do óleo essencial de Ocimum gratissimum durante o cultivo in vitro de folículos pré-antrais bovinos. 2020. 98 f. Dissertação (Mestrado em Biotecnologia), Programa de Pós-Graduação em Biotecnologia - Campus de Sobral, Universidade Federal do Ceará, Sobral, 2020.
http://www.repositorio.ufc.br/handle/riufc/53680
identifier_str_mv Vasconcelos.E.M. Avaliação dos efeitos do eugenol e do óleo essencial de Ocimum gratissimum durante o cultivo in vitro de folículos pré-antrais bovinos. 2020. 98 f. Dissertação (Mestrado em Biotecnologia), Programa de Pós-Graduação em Biotecnologia - Campus de Sobral, Universidade Federal do Ceará, Sobral, 2020.
url http://www.repositorio.ufc.br/handle/riufc/53680
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
dc.source.none.fl_str_mv reponame:Repositório Institucional da Universidade Federal do Ceará (UFC)
instname:Universidade Federal do Ceará (UFC)
instacron:UFC
instname_str Universidade Federal do Ceará (UFC)
instacron_str UFC
institution UFC
reponame_str Repositório Institucional da Universidade Federal do Ceará (UFC)
collection Repositório Institucional da Universidade Federal do Ceará (UFC)
repository.name.fl_str_mv Repositório Institucional da Universidade Federal do Ceará (UFC) - Universidade Federal do Ceará (UFC)
repository.mail.fl_str_mv bu@ufc.br || repositorio@ufc.br
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