A angiotensina II regula a esteroidogênese nas células da teca bovina?

Detalhes bibliográficos
Autor(a) principal: Rigo, Melânia Lazzari
Data de Publicação: 2014
Tipo de documento: Dissertação
Idioma: por
Título da fonte: Manancial - Repositório Digital da UFSM
Texto Completo: http://repositorio.ufsm.br/handle/1/10169
Resumo: Many studies have been developed to characterize the function of angiotensin-renin system (ARS) in the female reproductive organs. Evidences from the literature have pointed a relevant role of angiotensin II (Ang II) in mammals, through its type 2 receptor (AT2) in oocyte maturation as in ovulation. Nevertheless, the participation of Ang II in other important reproductive features such as steroidogenesis has not been fully clarified. Therefore, the main objective of this work was to detect in vitro the steroidogenic effects of Ang II in theca cells. For that, bovine theca cells were obtained from follicles (larger than 8mm of diameter) collected from a local abattoir and submitted to different treatments in a sequence of experiments. In Experiment 1, Ang II was added to LH-treated (10 ng/ml) theca cells. Experiment 2 employed Ang II, in different concentrations, in addition to insulin (100 ng ̸ml) and LH (100 ng ̸ml). Experiment 3 explored the effects of an Ang II antagonist (saralasin) in theca cells co-stimulated by insulin and LH (both at 100 ng ̸ml). After 24 hours, culture media were collected and evaluated for testosterone and androstenedione levels measured by high performance liquid chromatography (HPLC). In parallel, gene expression of key steroidogenic enzymes and proteins, respectively, HSD3B2, CYP11A1 e CYP17A1 and STAR were accessed by qRT-PCR, with exception of experiment 1, in which only CYP17A1 was evaluated. Overall, absence of Ang II action was observed in all Ang II doses evaluated. Despite the difference in gene expression for CYP17A1 against controls in experiment 1, neither an increase in androgens levels nor a negative impact of saralasin were detected. Although very important for oocyte maturation and the ovulation, Ang II seems not influence androgen production by theca cells in vitro. In conclusion our results do not support the role for Ang II in thecal steroidogenesis, at least in bovine, as the primary hypothesis of the study.
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spelling A angiotensina II regula a esteroidogênese nas células da teca bovina?Does angiotensin II regulate steroidogenesis in the theca cells in cattle?RASAngiotensina IILHEnzimas esteroidogênicasOvárioRASAngiotensin IILHSteroidogenic enzymesOvaryCNPQ::CIENCIAS AGRARIAS::MEDICINA VETERINARIAMany studies have been developed to characterize the function of angiotensin-renin system (ARS) in the female reproductive organs. Evidences from the literature have pointed a relevant role of angiotensin II (Ang II) in mammals, through its type 2 receptor (AT2) in oocyte maturation as in ovulation. Nevertheless, the participation of Ang II in other important reproductive features such as steroidogenesis has not been fully clarified. Therefore, the main objective of this work was to detect in vitro the steroidogenic effects of Ang II in theca cells. For that, bovine theca cells were obtained from follicles (larger than 8mm of diameter) collected from a local abattoir and submitted to different treatments in a sequence of experiments. In Experiment 1, Ang II was added to LH-treated (10 ng/ml) theca cells. Experiment 2 employed Ang II, in different concentrations, in addition to insulin (100 ng ̸ml) and LH (100 ng ̸ml). Experiment 3 explored the effects of an Ang II antagonist (saralasin) in theca cells co-stimulated by insulin and LH (both at 100 ng ̸ml). After 24 hours, culture media were collected and evaluated for testosterone and androstenedione levels measured by high performance liquid chromatography (HPLC). In parallel, gene expression of key steroidogenic enzymes and proteins, respectively, HSD3B2, CYP11A1 e CYP17A1 and STAR were accessed by qRT-PCR, with exception of experiment 1, in which only CYP17A1 was evaluated. Overall, absence of Ang II action was observed in all Ang II doses evaluated. Despite the difference in gene expression for CYP17A1 against controls in experiment 1, neither an increase in androgens levels nor a negative impact of saralasin were detected. Although very important for oocyte maturation and the ovulation, Ang II seems not influence androgen production by theca cells in vitro. In conclusion our results do not support the role for Ang II in thecal steroidogenesis, at least in bovine, as the primary hypothesis of the study.Coordenação de Aperfeiçoamento de Pessoal de Nível SuperiorDiversos estudos vêm sendo desenvolvidos para caracterizar o sistema renina angiotensina (RAS) no aparelho reprodutivo feminino. Evidências da literatura apontam um importante papel da angiotensina II (Ang II), via receptor tipo 2 (AT2), tanto na maturação dos oócitos quanto na ovulação em mamíferos. No entanto, a participação da Ang II em outros aspectos reprodutivos importantes, como a esteroidogênese, ainda não foi completamente elucidada. Desta forma, o objetivo deste trabalho foi verificar o efeito in vitro da Ang II nas células da teca cultivadas. Para isso, células da teca bovina foram obtidas de folículos com mais de 8mm de diâmetro de ovários oriundos de abatedouro local e submetidas a diferentes tratamentos em uma sequência de experimentos. No experimento 1, Ang II foi adicionada a células da teca tratadas com LH na dose de 10 ng/ml. No experimento 2, foi utilizada Ang II em diferentes concentrações em adição ao tratamento com insulina (100 ng ̸ml) e LH (100 ng ̸ml). O experimento 3, explorou o possível efeito de um antagonista da Ang II (saralasina) em células da teca co-estimuladas com insulina e LH (ambos em 100 ng ̸ml). Após 24 horas, o meio de cultura foi coletado e avaliado para verificação dos níveis de testosterona e androstenediona aferidos pela técnica de cromatografia líquida de alta performance (HPLC). Em paralelo, a expressão gênica de enzimas e proteínas chaves na esteroidogênese, respectivamente, HSD3B2, CYP11A1 e CYP17A1 e STAR, foram avaliadas por qRT-PCR, com exceção do experimento 1 onde somente a CYP17A1 foi estudada. De maneira geral, não foi observada uma ação da Ang II nas doses utilizadas. Apesar de uma diferença na expressão de CYP17A1 ter sido verificada em relação aos controles, nem o aumento dos níveis de androgênios ou um impacto negativo pelo uso de saralasina foram detectados. Embora reconhecida como muito importante para maturação oocitária e ovulação, a Ang II parece não influenciar a produção de androgênios in vitro. Em conclusão, nossos resultados não demonstraram um papel da Ang II na esteroidogênese tecal, pelo menos na espécie bovina, ao contrário da hipótese original deste estudo.Universidade Federal de Santa MariaBRMedicina VeterináriaUFSMPrograma de Pós-Graduação em Medicina VeterináriaGonçalves, Paulo Bayard Diashttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4781845H4Antoniazzi, Alfredo Quiteshttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4162414A9Nóbrega Junior, Jandui Escarião dahttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4757536T3Rigo, Melânia Lazzari2014-10-012014-10-012014-02-17info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisapplication/pdfapplication/pdfRIGO, Melânia Lazzari. DOES ANGIOTENSIN II REGULATE STEROIDOGENESIS IN THE THECA CELLS IN CATTLE?. 2014. 42 f. Dissertação (Mestrado em Medicina Veterinária) - Universidade Federal de Santa Maria, Santa Maria, 2014.http://repositorio.ufsm.br/handle/1/10169porinfo:eu-repo/semantics/openAccessreponame:Manancial - Repositório Digital da UFSMinstname:Universidade Federal de Santa Maria (UFSM)instacron:UFSM2017-12-02T15:41:53Zoai:repositorio.ufsm.br:1/10169Biblioteca Digital de Teses e Dissertaçõeshttps://repositorio.ufsm.br/ONGhttps://repositorio.ufsm.br/oai/requestatendimento.sib@ufsm.br||tedebc@gmail.comopendoar:2017-12-02T15:41:53Manancial - Repositório Digital da UFSM - Universidade Federal de Santa Maria (UFSM)false
dc.title.none.fl_str_mv A angiotensina II regula a esteroidogênese nas células da teca bovina?
Does angiotensin II regulate steroidogenesis in the theca cells in cattle?
title A angiotensina II regula a esteroidogênese nas células da teca bovina?
spellingShingle A angiotensina II regula a esteroidogênese nas células da teca bovina?
Rigo, Melânia Lazzari
RAS
Angiotensina II
LH
Enzimas esteroidogênicas
Ovário
RAS
Angiotensin II
LH
Steroidogenic enzymes
Ovary
CNPQ::CIENCIAS AGRARIAS::MEDICINA VETERINARIA
title_short A angiotensina II regula a esteroidogênese nas células da teca bovina?
title_full A angiotensina II regula a esteroidogênese nas células da teca bovina?
title_fullStr A angiotensina II regula a esteroidogênese nas células da teca bovina?
title_full_unstemmed A angiotensina II regula a esteroidogênese nas células da teca bovina?
title_sort A angiotensina II regula a esteroidogênese nas células da teca bovina?
author Rigo, Melânia Lazzari
author_facet Rigo, Melânia Lazzari
author_role author
dc.contributor.none.fl_str_mv Gonçalves, Paulo Bayard Dias
http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4781845H4
Antoniazzi, Alfredo Quites
http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4162414A9
Nóbrega Junior, Jandui Escarião da
http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4757536T3
dc.contributor.author.fl_str_mv Rigo, Melânia Lazzari
dc.subject.por.fl_str_mv RAS
Angiotensina II
LH
Enzimas esteroidogênicas
Ovário
RAS
Angiotensin II
LH
Steroidogenic enzymes
Ovary
CNPQ::CIENCIAS AGRARIAS::MEDICINA VETERINARIA
topic RAS
Angiotensina II
LH
Enzimas esteroidogênicas
Ovário
RAS
Angiotensin II
LH
Steroidogenic enzymes
Ovary
CNPQ::CIENCIAS AGRARIAS::MEDICINA VETERINARIA
description Many studies have been developed to characterize the function of angiotensin-renin system (ARS) in the female reproductive organs. Evidences from the literature have pointed a relevant role of angiotensin II (Ang II) in mammals, through its type 2 receptor (AT2) in oocyte maturation as in ovulation. Nevertheless, the participation of Ang II in other important reproductive features such as steroidogenesis has not been fully clarified. Therefore, the main objective of this work was to detect in vitro the steroidogenic effects of Ang II in theca cells. For that, bovine theca cells were obtained from follicles (larger than 8mm of diameter) collected from a local abattoir and submitted to different treatments in a sequence of experiments. In Experiment 1, Ang II was added to LH-treated (10 ng/ml) theca cells. Experiment 2 employed Ang II, in different concentrations, in addition to insulin (100 ng ̸ml) and LH (100 ng ̸ml). Experiment 3 explored the effects of an Ang II antagonist (saralasin) in theca cells co-stimulated by insulin and LH (both at 100 ng ̸ml). After 24 hours, culture media were collected and evaluated for testosterone and androstenedione levels measured by high performance liquid chromatography (HPLC). In parallel, gene expression of key steroidogenic enzymes and proteins, respectively, HSD3B2, CYP11A1 e CYP17A1 and STAR were accessed by qRT-PCR, with exception of experiment 1, in which only CYP17A1 was evaluated. Overall, absence of Ang II action was observed in all Ang II doses evaluated. Despite the difference in gene expression for CYP17A1 against controls in experiment 1, neither an increase in androgens levels nor a negative impact of saralasin were detected. Although very important for oocyte maturation and the ovulation, Ang II seems not influence androgen production by theca cells in vitro. In conclusion our results do not support the role for Ang II in thecal steroidogenesis, at least in bovine, as the primary hypothesis of the study.
publishDate 2014
dc.date.none.fl_str_mv 2014-10-01
2014-10-01
2014-02-17
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 RIGO, Melânia Lazzari. DOES ANGIOTENSIN II REGULATE STEROIDOGENESIS IN THE THECA CELLS IN CATTLE?. 2014. 42 f. Dissertação (Mestrado em Medicina Veterinária) - Universidade Federal de Santa Maria, Santa Maria, 2014.
http://repositorio.ufsm.br/handle/1/10169
identifier_str_mv RIGO, Melânia Lazzari. DOES ANGIOTENSIN II REGULATE STEROIDOGENESIS IN THE THECA CELLS IN CATTLE?. 2014. 42 f. Dissertação (Mestrado em Medicina Veterinária) - Universidade Federal de Santa Maria, Santa Maria, 2014.
url http://repositorio.ufsm.br/handle/1/10169
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
Medicina Veterinária
UFSM
Programa de Pós-Graduação em Medicina Veterinária
publisher.none.fl_str_mv Universidade Federal de Santa Maria
BR
Medicina Veterinária
UFSM
Programa de Pós-Graduação em Medicina Veterinária
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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