Uso de inibidores de proteína quinase C e fosfolipase A2 em células endometriais bovinas tratadas com estradiol e ionóforo de cálcio
Autor(a) principal: | |
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Data de Publicação: | 2021 |
Outros Autores: | , , , , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Brazilian Journal of Veterinary Research and Animal Science |
Texto Completo: | https://www.revistas.usp.br/bjvras/article/view/174355 |
Resumo: | The release of endometrial prostaglandin-F2α (PGF2α) in bovine females can be induced in vivo by estradiol (E2). However, its role in this mechanism has not been clarified. We hypothesized that E2 stimulates the activity and abundance of protein kinase C (PKC) and phospholipase A2 (PLA2). Our objective in this study was to analyze the effects of PKC and PLA2 inhibitors on PGF2α synthesis induced by E2 and calcium ionophore (CI) in bovine endometrial cells (BEND cells; Experiment 1). Additionally, we evaluated the abundance of PKC and PLA2 in endometrial explants of cows treated or not with E2 17 days after estrus (D17, D0 = estrus; Experiment 2). In Experiment 1, BEND cells were submitted to a PKC inhibitor (10 μM of C25H24N4O2; bisindolylmaleimide I, or BIS I), a PLA2 inhibitor (20 μM of arachydoniltrifluoromethane or AACOCF3), or none. The BEND cells were subsequently treated with E2 and CI, and PGF2α concentrations were measured in the culture medium through radioimmunoassay. For DIF-12 (PGF2α concentration 12 h after treatment subtracted from PGF2α concentration at hour 0), no PKC inhibitor effect was observed (P= 0.2709). However, DIF-12 was lower (P < 0.05) for groups treated with the PLA2 inhibitor and PLA2 inhibitor + CI + E2 groups than the control and CI + E2 groups. Thus, AACOCF3 was an efficient PLA2 inhibitor in the BEND cells culture system, and E2 did not stimulate the synthesis of PKC and PLA2. In Experiment 2, cyclic Nellore heifers received none (n = 5) or 3 mg (n = 6) of 17β-E2 on D17 and were slaughtered 2 h after administration. The abundance of PKC and PLA2 in the endometrial tissue was evaluated using Western blotting analysis. No E2 effect was observed on PKC (P = 0.08) and PLA2 (P = 0.56). We concluded that E2 did not stimulate the activity and abundance of PKC and PLA2. |
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Brazilian Journal of Veterinary Research and Animal Science |
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Uso de inibidores de proteína quinase C e fosfolipase A2 em células endometriais bovinas tratadas com estradiol e ionóforo de cálcioUse of protein kinase C and phospholipase A2 inhibitors in bovine endometrial cells treated with estradiol and calcium ionophoreEstradiolPGF2αPKCPLA2Células BENDEstradiolPGF2αPKCPLA2BEND cellsThe release of endometrial prostaglandin-F2α (PGF2α) in bovine females can be induced in vivo by estradiol (E2). However, its role in this mechanism has not been clarified. We hypothesized that E2 stimulates the activity and abundance of protein kinase C (PKC) and phospholipase A2 (PLA2). Our objective in this study was to analyze the effects of PKC and PLA2 inhibitors on PGF2α synthesis induced by E2 and calcium ionophore (CI) in bovine endometrial cells (BEND cells; Experiment 1). Additionally, we evaluated the abundance of PKC and PLA2 in endometrial explants of cows treated or not with E2 17 days after estrus (D17, D0 = estrus; Experiment 2). In Experiment 1, BEND cells were submitted to a PKC inhibitor (10 μM of C25H24N4O2; bisindolylmaleimide I, or BIS I), a PLA2 inhibitor (20 μM of arachydoniltrifluoromethane or AACOCF3), or none. The BEND cells were subsequently treated with E2 and CI, and PGF2α concentrations were measured in the culture medium through radioimmunoassay. For DIF-12 (PGF2α concentration 12 h after treatment subtracted from PGF2α concentration at hour 0), no PKC inhibitor effect was observed (P= 0.2709). However, DIF-12 was lower (P < 0.05) for groups treated with the PLA2 inhibitor and PLA2 inhibitor + CI + E2 groups than the control and CI + E2 groups. Thus, AACOCF3 was an efficient PLA2 inhibitor in the BEND cells culture system, and E2 did not stimulate the synthesis of PKC and PLA2. In Experiment 2, cyclic Nellore heifers received none (n = 5) or 3 mg (n = 6) of 17β-E2 on D17 and were slaughtered 2 h after administration. The abundance of PKC and PLA2 in the endometrial tissue was evaluated using Western blotting analysis. No E2 effect was observed on PKC (P = 0.08) and PLA2 (P = 0.56). We concluded that E2 did not stimulate the activity and abundance of PKC and PLA2.A liberação endometrial de prostaglandina-F2α (PGF2α) em fêmeas bovinas pode ser induzida in vivo pelo estradiol (E2). Entretanto o seu mecanismo de ação ainda não foi bem esclarecido. Nossa hipótese é que o E2 estimula a atividade e a abundância da proteína quinase C (PKC) e da fosfolipase A2 (PLA2). Nosso objetivo com este estudo foi analisar os efeitos de inibidores de PKC e PLA2 na síntese de PGF2α induzida por E2 e ionóforo de cálcio (CI) em células endometriais bovinas (células BEND; Experimento 1). Adicionalmente, nós avaliamos a abundância de PKC e PLA2 em explantes endometriais de vacas tratadas com ou sem E2 17 dias após o estro (D17, D0 = estro; Experimento 2). No Experimento 1, células BEND foram submetidas ao inibidor de PKC (10 μM de C25H24N4O2; bisindolylmaleimide I, ou BIS I), e ao inibidor de PLA2 (20 μM de arachydoniltrifluoromethane ou AACOCF3) ou a nenhum inibidor. As células BEND foram subsequentemente tratadas com E2 e CI e concentrações de PGF2α foram mensuradas no meio de cultura por radioimunoenssaio. Para DIF-12 (concentração de PGF2α 12 horas depois do tratamento, subtraída da concentração de PGF2α na hora 0), não foi observado efeito do inibidor de PKC (P = 0.2709). Entretanto DIF-12 foi menor (P < 0.05) nos grupos tratados com inibidor de PLA2 e inibidor de PLA2 + CI + E2 quando comparados com o grupo controle e o grupo CI + E2. O AACOCF3 foi um eficiente inibidor de PLA2 em sistema de cultura de células BEND e o E2 não estimulou a síntese de PKC e PLA2. No Experimento 2, novilhas Nelore cíclicas receberam 3 mg de 17β-E2 (n = 6) ou nenhum tratamento (n = 5) no D17 e foram abatidas duas horas depois da administração dos tratamentos. A quantidade de PKC e PLA2 no tecido endometrial foi avaliada pela técnica de Western Blotting. Não foi observado efeito do E2 sobre a PKC (P= 0.08) e nem sobre a PLA2 (P= 0.56). Conclui-se que o E2 não estimulou a atividade e abundância de PKC e PLA2.Universidade de São Paulo. Faculdade de Medicina Veterinária e Zootecnia2021-05-07info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionapplication/pdfhttps://www.revistas.usp.br/bjvras/article/view/17435510.11606/issn.1678-4456.bjvras.2021.174355Brazilian Journal of Veterinary Research and Animal Science; Vol. 58 (2021); e174355Brazilian Journal of Veterinary Research and Animal Science; Vol. 58 (2021); e174355Brazilian Journal of Veterinary Research and Animal Science; v. 58 (2021); e174355Brazilian Journal of Veterinary Research and Animal Science; V. 58 (2021); e1743551678-44561413-9596reponame:Brazilian Journal of Veterinary Research and Animal Scienceinstname:Faculdade de Medicina Veterinária e Zootecnia da Universidade de São Paulo (FMVZ-USP)instacron:USPenghttps://www.revistas.usp.br/bjvras/article/view/174355/171549Copyright (c) 2021 Brazilian Journal of Veterinary Research and Animal Sciencehttps://creativecommons.org/licenses/by-nc-sa/4.0info:eu-repo/semantics/openAccessMaldonado, Mariângela Bueno CordeiroHenry, Francine Messias CiríacoFerreira, Teissiane Fernanda de VasconcelosMello, Barbara PifferoBinelli, MarioMembrive, Claudia Maria Bertan2021-03-23T19:37:57Zoai:revistas.usp.br:article/174355Revistahttps://www.revistas.usp.br/bjvrasPUBhttps://www.revistas.usp.br/bjvras/oaibjvras@usp.br1413-95961413-9596opendoar:https://www.revistas.usp.br/bjvras/index2023-01-12T16:44:07.268410Brazilian Journal of Veterinary Research and Animal Science - Faculdade de Medicina Veterinária e Zootecnia da Universidade de São Paulo (FMVZ-USP)false |
dc.title.none.fl_str_mv |
Uso de inibidores de proteína quinase C e fosfolipase A2 em células endometriais bovinas tratadas com estradiol e ionóforo de cálcio Use of protein kinase C and phospholipase A2 inhibitors in bovine endometrial cells treated with estradiol and calcium ionophore |
title |
Uso de inibidores de proteína quinase C e fosfolipase A2 em células endometriais bovinas tratadas com estradiol e ionóforo de cálcio |
spellingShingle |
Uso de inibidores de proteína quinase C e fosfolipase A2 em células endometriais bovinas tratadas com estradiol e ionóforo de cálcio Maldonado, Mariângela Bueno Cordeiro Estradiol PGF2α PKC PLA2 Células BEND Estradiol PGF2α PKC PLA2 BEND cells |
title_short |
Uso de inibidores de proteína quinase C e fosfolipase A2 em células endometriais bovinas tratadas com estradiol e ionóforo de cálcio |
title_full |
Uso de inibidores de proteína quinase C e fosfolipase A2 em células endometriais bovinas tratadas com estradiol e ionóforo de cálcio |
title_fullStr |
Uso de inibidores de proteína quinase C e fosfolipase A2 em células endometriais bovinas tratadas com estradiol e ionóforo de cálcio |
title_full_unstemmed |
Uso de inibidores de proteína quinase C e fosfolipase A2 em células endometriais bovinas tratadas com estradiol e ionóforo de cálcio |
title_sort |
Uso de inibidores de proteína quinase C e fosfolipase A2 em células endometriais bovinas tratadas com estradiol e ionóforo de cálcio |
author |
Maldonado, Mariângela Bueno Cordeiro |
author_facet |
Maldonado, Mariângela Bueno Cordeiro Henry, Francine Messias Ciríaco Ferreira, Teissiane Fernanda de Vasconcelos Mello, Barbara Piffero Binelli, Mario Membrive, Claudia Maria Bertan |
author_role |
author |
author2 |
Henry, Francine Messias Ciríaco Ferreira, Teissiane Fernanda de Vasconcelos Mello, Barbara Piffero Binelli, Mario Membrive, Claudia Maria Bertan |
author2_role |
author author author author author |
dc.contributor.author.fl_str_mv |
Maldonado, Mariângela Bueno Cordeiro Henry, Francine Messias Ciríaco Ferreira, Teissiane Fernanda de Vasconcelos Mello, Barbara Piffero Binelli, Mario Membrive, Claudia Maria Bertan |
dc.subject.por.fl_str_mv |
Estradiol PGF2α PKC PLA2 Células BEND Estradiol PGF2α PKC PLA2 BEND cells |
topic |
Estradiol PGF2α PKC PLA2 Células BEND Estradiol PGF2α PKC PLA2 BEND cells |
description |
The release of endometrial prostaglandin-F2α (PGF2α) in bovine females can be induced in vivo by estradiol (E2). However, its role in this mechanism has not been clarified. We hypothesized that E2 stimulates the activity and abundance of protein kinase C (PKC) and phospholipase A2 (PLA2). Our objective in this study was to analyze the effects of PKC and PLA2 inhibitors on PGF2α synthesis induced by E2 and calcium ionophore (CI) in bovine endometrial cells (BEND cells; Experiment 1). Additionally, we evaluated the abundance of PKC and PLA2 in endometrial explants of cows treated or not with E2 17 days after estrus (D17, D0 = estrus; Experiment 2). In Experiment 1, BEND cells were submitted to a PKC inhibitor (10 μM of C25H24N4O2; bisindolylmaleimide I, or BIS I), a PLA2 inhibitor (20 μM of arachydoniltrifluoromethane or AACOCF3), or none. The BEND cells were subsequently treated with E2 and CI, and PGF2α concentrations were measured in the culture medium through radioimmunoassay. For DIF-12 (PGF2α concentration 12 h after treatment subtracted from PGF2α concentration at hour 0), no PKC inhibitor effect was observed (P= 0.2709). However, DIF-12 was lower (P < 0.05) for groups treated with the PLA2 inhibitor and PLA2 inhibitor + CI + E2 groups than the control and CI + E2 groups. Thus, AACOCF3 was an efficient PLA2 inhibitor in the BEND cells culture system, and E2 did not stimulate the synthesis of PKC and PLA2. In Experiment 2, cyclic Nellore heifers received none (n = 5) or 3 mg (n = 6) of 17β-E2 on D17 and were slaughtered 2 h after administration. The abundance of PKC and PLA2 in the endometrial tissue was evaluated using Western blotting analysis. No E2 effect was observed on PKC (P = 0.08) and PLA2 (P = 0.56). We concluded that E2 did not stimulate the activity and abundance of PKC and PLA2. |
publishDate |
2021 |
dc.date.none.fl_str_mv |
2021-05-07 |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/article info:eu-repo/semantics/publishedVersion |
format |
article |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
https://www.revistas.usp.br/bjvras/article/view/174355 10.11606/issn.1678-4456.bjvras.2021.174355 |
url |
https://www.revistas.usp.br/bjvras/article/view/174355 |
identifier_str_mv |
10.11606/issn.1678-4456.bjvras.2021.174355 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
https://www.revistas.usp.br/bjvras/article/view/174355/171549 |
dc.rights.driver.fl_str_mv |
Copyright (c) 2021 Brazilian Journal of Veterinary Research and Animal Science https://creativecommons.org/licenses/by-nc-sa/4.0 info:eu-repo/semantics/openAccess |
rights_invalid_str_mv |
Copyright (c) 2021 Brazilian Journal of Veterinary Research and Animal Science https://creativecommons.org/licenses/by-nc-sa/4.0 |
eu_rights_str_mv |
openAccess |
dc.format.none.fl_str_mv |
application/pdf |
dc.publisher.none.fl_str_mv |
Universidade de São Paulo. Faculdade de Medicina Veterinária e Zootecnia |
publisher.none.fl_str_mv |
Universidade de São Paulo. Faculdade de Medicina Veterinária e Zootecnia |
dc.source.none.fl_str_mv |
Brazilian Journal of Veterinary Research and Animal Science; Vol. 58 (2021); e174355 Brazilian Journal of Veterinary Research and Animal Science; Vol. 58 (2021); e174355 Brazilian Journal of Veterinary Research and Animal Science; v. 58 (2021); e174355 Brazilian Journal of Veterinary Research and Animal Science; V. 58 (2021); e174355 1678-4456 1413-9596 reponame:Brazilian Journal of Veterinary Research and Animal Science instname:Faculdade de Medicina Veterinária e Zootecnia da Universidade de São Paulo (FMVZ-USP) instacron:USP |
instname_str |
Faculdade de Medicina Veterinária e Zootecnia da Universidade de São Paulo (FMVZ-USP) |
instacron_str |
USP |
institution |
USP |
reponame_str |
Brazilian Journal of Veterinary Research and Animal Science |
collection |
Brazilian Journal of Veterinary Research and Animal Science |
repository.name.fl_str_mv |
Brazilian Journal of Veterinary Research and Animal Science - Faculdade de Medicina Veterinária e Zootecnia da Universidade de São Paulo (FMVZ-USP) |
repository.mail.fl_str_mv |
bjvras@usp.br |
_version_ |
1797051567958917120 |