Síntese e estudo da atividade antinociceptiva de Pep13, peptídeo derivado de PnTx4(6-1), uma toxina da peçonha da aranha Phoneutria nigriventer

Detalhes bibliográficos
Autor(a) principal: Bruna Luiza Emerich Magalhães
Data de Publicação: 2017
Tipo de documento: Tese
Idioma: por
Título da fonte: Repositório Institucional da UFMG
Texto Completo: http://hdl.handle.net/1843/35463
Resumo: The venom of the “armed” spider Phoneutria nigriventer has several biologically active peptides, one of them the toxin PnTx4(6-1) is a peptide composed of 48 amino acid residues and molecular mass of 5.2 kDa. PnTx4(6-1), also named δ-ctenitoxin-Pn1a, was initially described as a insecticidal neurotoxin, that binds to site 3 of sodium channels, in nerve cord synaptosomes of cockroach. When intracerebral injected in (30 μg) in mice, PnTx4(6-1) caused no apparent toxicity, moreover this toxin did not affect sodium channels of skeletal muscle (rSKM1) and brain (rBIIa), both of rats. Subsequently, we demonstrated that PnTx4(6-1) has antinociceptive effect in three pain models: i) inflammatory, evoked by carrageenan; ii) nociceptive, evoked by prostaglandin E2 and iii) neuropathic, evoked by constriction of the sciatic nerve. Using diverse receptors antagonists, we verified that the cannabinoid system, via the CB1 receptor, and the opioid system, through the μ and δ receptors, are both involved in the antinociceptive effect of PnTx4(6-1). In the present work, based on PnTx4(6-1) amino acid sequence we proposed and synthesized a linear peptide, with 13 amino acid residues named Pep13, based on the amino acid sequence of PnTx4(6-1) in order to try to reproduce or increase the analgesic effect of the toxin, with a simpler molecule. We verified, similarly to the toxin, the peptide has antinociceptive activity, when intrathecally administered, and this effect is also related to the cannabinoid and opioid systems. In addition, when the peripheral effect was evaluated, by intraplantar admistration, Pep13 was able to reverse the hyperalgesic threshold evoked by prostaglandin E2. Still related to the peripheral antinociceptive effect, the participation of the opioid system was observed, since the nonspecific antagonist (naloxone) was able to partially prevent the effect of Pep13. Regarding the cannabinoid system, by using specific antagonists, the participation of CB1 receptors was observed. Differently from the native toxin, which has a high toxicity to insects, the peptide caused no apparent effect when administered in flies. In conclusion, the synthetic peptide Pep13, which has 35 amino acid residues less than the native toxin PnTx4(6-1), reproduces the antinociceptive effects presented therein, without presenting however insect toxicity and may become a good model for new analgesics.
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spelling Síntese e estudo da atividade antinociceptiva de Pep13, peptídeo derivado de PnTx4(6-1), uma toxina da peçonha da aranha Phoneutria nigriventerPhoneutria nigriventerPnTx4(6-1)AntinocicepçãoPep13AranhasVenenos de aranhaPhoneutria nigriventerThe venom of the “armed” spider Phoneutria nigriventer has several biologically active peptides, one of them the toxin PnTx4(6-1) is a peptide composed of 48 amino acid residues and molecular mass of 5.2 kDa. PnTx4(6-1), also named δ-ctenitoxin-Pn1a, was initially described as a insecticidal neurotoxin, that binds to site 3 of sodium channels, in nerve cord synaptosomes of cockroach. When intracerebral injected in (30 μg) in mice, PnTx4(6-1) caused no apparent toxicity, moreover this toxin did not affect sodium channels of skeletal muscle (rSKM1) and brain (rBIIa), both of rats. Subsequently, we demonstrated that PnTx4(6-1) has antinociceptive effect in three pain models: i) inflammatory, evoked by carrageenan; ii) nociceptive, evoked by prostaglandin E2 and iii) neuropathic, evoked by constriction of the sciatic nerve. Using diverse receptors antagonists, we verified that the cannabinoid system, via the CB1 receptor, and the opioid system, through the μ and δ receptors, are both involved in the antinociceptive effect of PnTx4(6-1). In the present work, based on PnTx4(6-1) amino acid sequence we proposed and synthesized a linear peptide, with 13 amino acid residues named Pep13, based on the amino acid sequence of PnTx4(6-1) in order to try to reproduce or increase the analgesic effect of the toxin, with a simpler molecule. We verified, similarly to the toxin, the peptide has antinociceptive activity, when intrathecally administered, and this effect is also related to the cannabinoid and opioid systems. In addition, when the peripheral effect was evaluated, by intraplantar admistration, Pep13 was able to reverse the hyperalgesic threshold evoked by prostaglandin E2. Still related to the peripheral antinociceptive effect, the participation of the opioid system was observed, since the nonspecific antagonist (naloxone) was able to partially prevent the effect of Pep13. Regarding the cannabinoid system, by using specific antagonists, the participation of CB1 receptors was observed. Differently from the native toxin, which has a high toxicity to insects, the peptide caused no apparent effect when administered in flies. In conclusion, the synthetic peptide Pep13, which has 35 amino acid residues less than the native toxin PnTx4(6-1), reproduces the antinociceptive effects presented therein, without presenting however insect toxicity and may become a good model for new analgesics.A peçonha da aranha "armadeira" Phoneutria nigriventer possui diversos componentes biologicamente ativos, dentre eles a toxina PnTx4(6-1), um peptídeo composto por 48 resíduos de aminoácidos e massa molecular de 5,2 kDa. PnTx4(6-1), também denominada δ-ctenitoxin-Pn1a, foi inicialmente descrita como uma neurotoxina inseticida que se liga ao sítio 3 de canais para sódio em sinaptossomas de corda nervosa de barata. Quando injetada por via intracerebroventricular em camundongos (30 μg), PnTx4(6-1) não provocou toxicidade aparente, além disso, esta toxina não apresentou efeito em canais para sódio de músculo esquelético (rSKM1) e de cérebro (rBIIA) de ratos. Posteriormente demonstramos que PnTx4(6-1) possui efeito antinociceptivo frente a três modelos de dor: i) inflamatório, evocado por carragenina; ii) nociceptivo, evocado por prostaglandina E2 e iii) neuropático, evocado por constrição do nervo ciático. Utilizando-se antagonistas verificamos que o sistema canabinoide, via receptor CB1, e o sistema opioide, via receptores μ e δ, estão envolvidos no efeito antinociceptivo evocado por PnTx4(6-1). No presente trabalho baseado na sequencia de aminoácidos de PnTx4(6-1), propusemos e sintetizamos um peptídeo linear, de 13 resíduos de aminoácidos denominado Pep13, na tentativa de reproduzir ou aumentar os efeitos antinociceptivos da toxina nativa. Verificamos que, assim como a toxina nativa, Pep13 apresenta ação antinociceptiva quando administrado por via intratecal e este efeito, também, relaciona-se aos sistemas canabinoide e opioide. Além disto, quando o efeito periférico foi avaliado, por administração intraplantar, Pep13 foi capaz de reverter o limiar hiperalgésico evocado por prostaglandina E2. Relacionada, ainda, ao efeito antinociceptivo periférico foi observada a participação do sistema opioide, uma vez que o antagonista não específico (naloxona) foi capaz de prevenir parcialmente o efeito de Pep13. Com relação ao sistema canabinoide, com a utilização de antagonistas específicos, observou-se a participação dos receptores CB1. Diferentemente da inseto-toxina nativa, Pep13 não causou nenhum efeito aparente quando administrado em moscas. Em conclusão, o peptídeo sintético Pep13, cuja sequência possui 35 resíduos de aminoácidos a menos que a toxina nativa PnTx4(6-1), reproduz os efeitos antinociceptivos apresentados pela mesma, sem entretanto, apresentar toxicidade para insetos e podendo assim, se tornar um bom modelo para novos analgésicos.CNPq - Conselho Nacional de Desenvolvimento Científico e TecnológicoFAPEMIG - Fundação de Amparo à Pesquisa do Estado de Minas GeraisCAPES - Coordenação de Aperfeiçoamento de Pessoal de Nível SuperiorUniversidade Federal de Minas GeraisBrasilICB - DEPARTAMENTO DE BIOQUÍMICA E IMUNOLOGIAPrograma de Pós-Graduação em Bioquímica e ImunologiaUFMGMaria Elena de Lima Perez Garciahttp://lattes.cnpq.br/5642915270924367Igor Dimitri Gama DuarteMarie France Martin-EauclaireMariana Torquato Quezado MagalhãesDaniele Cristina de AguiarCélio José de Castro JúniorMárcia Renata MortariBruna Luiza Emerich Magalhães2021-03-29T13:05:51Z2021-03-29T13:05:51Z2017-05-15info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/doctoralThesisapplication/pdfhttp://hdl.handle.net/1843/35463porhttp://creativecommons.org/licenses/by-nc-nd/3.0/pt/info:eu-repo/semantics/openAccessreponame:Repositório Institucional da UFMGinstname:Universidade Federal de Minas Gerais (UFMG)instacron:UFMG2021-03-29T13:05:51Zoai:repositorio.ufmg.br:1843/35463Repositório InstitucionalPUBhttps://repositorio.ufmg.br/oairepositorio@ufmg.bropendoar:2021-03-29T13:05:51Repositório Institucional da UFMG - Universidade Federal de Minas Gerais (UFMG)false
dc.title.none.fl_str_mv Síntese e estudo da atividade antinociceptiva de Pep13, peptídeo derivado de PnTx4(6-1), uma toxina da peçonha da aranha Phoneutria nigriventer
title Síntese e estudo da atividade antinociceptiva de Pep13, peptídeo derivado de PnTx4(6-1), uma toxina da peçonha da aranha Phoneutria nigriventer
spellingShingle Síntese e estudo da atividade antinociceptiva de Pep13, peptídeo derivado de PnTx4(6-1), uma toxina da peçonha da aranha Phoneutria nigriventer
Bruna Luiza Emerich Magalhães
Phoneutria nigriventer
PnTx4(6-1)
Antinocicepção
Pep13
Aranhas
Venenos de aranha
Phoneutria nigriventer
title_short Síntese e estudo da atividade antinociceptiva de Pep13, peptídeo derivado de PnTx4(6-1), uma toxina da peçonha da aranha Phoneutria nigriventer
title_full Síntese e estudo da atividade antinociceptiva de Pep13, peptídeo derivado de PnTx4(6-1), uma toxina da peçonha da aranha Phoneutria nigriventer
title_fullStr Síntese e estudo da atividade antinociceptiva de Pep13, peptídeo derivado de PnTx4(6-1), uma toxina da peçonha da aranha Phoneutria nigriventer
title_full_unstemmed Síntese e estudo da atividade antinociceptiva de Pep13, peptídeo derivado de PnTx4(6-1), uma toxina da peçonha da aranha Phoneutria nigriventer
title_sort Síntese e estudo da atividade antinociceptiva de Pep13, peptídeo derivado de PnTx4(6-1), uma toxina da peçonha da aranha Phoneutria nigriventer
author Bruna Luiza Emerich Magalhães
author_facet Bruna Luiza Emerich Magalhães
author_role author
dc.contributor.none.fl_str_mv Maria Elena de Lima Perez Garcia
http://lattes.cnpq.br/5642915270924367
Igor Dimitri Gama Duarte
Marie France Martin-Eauclaire
Mariana Torquato Quezado Magalhães
Daniele Cristina de Aguiar
Célio José de Castro Júnior
Márcia Renata Mortari
dc.contributor.author.fl_str_mv Bruna Luiza Emerich Magalhães
dc.subject.por.fl_str_mv Phoneutria nigriventer
PnTx4(6-1)
Antinocicepção
Pep13
Aranhas
Venenos de aranha
Phoneutria nigriventer
topic Phoneutria nigriventer
PnTx4(6-1)
Antinocicepção
Pep13
Aranhas
Venenos de aranha
Phoneutria nigriventer
description The venom of the “armed” spider Phoneutria nigriventer has several biologically active peptides, one of them the toxin PnTx4(6-1) is a peptide composed of 48 amino acid residues and molecular mass of 5.2 kDa. PnTx4(6-1), also named δ-ctenitoxin-Pn1a, was initially described as a insecticidal neurotoxin, that binds to site 3 of sodium channels, in nerve cord synaptosomes of cockroach. When intracerebral injected in (30 μg) in mice, PnTx4(6-1) caused no apparent toxicity, moreover this toxin did not affect sodium channels of skeletal muscle (rSKM1) and brain (rBIIa), both of rats. Subsequently, we demonstrated that PnTx4(6-1) has antinociceptive effect in three pain models: i) inflammatory, evoked by carrageenan; ii) nociceptive, evoked by prostaglandin E2 and iii) neuropathic, evoked by constriction of the sciatic nerve. Using diverse receptors antagonists, we verified that the cannabinoid system, via the CB1 receptor, and the opioid system, through the μ and δ receptors, are both involved in the antinociceptive effect of PnTx4(6-1). In the present work, based on PnTx4(6-1) amino acid sequence we proposed and synthesized a linear peptide, with 13 amino acid residues named Pep13, based on the amino acid sequence of PnTx4(6-1) in order to try to reproduce or increase the analgesic effect of the toxin, with a simpler molecule. We verified, similarly to the toxin, the peptide has antinociceptive activity, when intrathecally administered, and this effect is also related to the cannabinoid and opioid systems. In addition, when the peripheral effect was evaluated, by intraplantar admistration, Pep13 was able to reverse the hyperalgesic threshold evoked by prostaglandin E2. Still related to the peripheral antinociceptive effect, the participation of the opioid system was observed, since the nonspecific antagonist (naloxone) was able to partially prevent the effect of Pep13. Regarding the cannabinoid system, by using specific antagonists, the participation of CB1 receptors was observed. Differently from the native toxin, which has a high toxicity to insects, the peptide caused no apparent effect when administered in flies. In conclusion, the synthetic peptide Pep13, which has 35 amino acid residues less than the native toxin PnTx4(6-1), reproduces the antinociceptive effects presented therein, without presenting however insect toxicity and may become a good model for new analgesics.
publishDate 2017
dc.date.none.fl_str_mv 2017-05-15
2021-03-29T13:05:51Z
2021-03-29T13:05:51Z
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
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format doctoralThesis
status_str publishedVersion
dc.identifier.uri.fl_str_mv http://hdl.handle.net/1843/35463
url http://hdl.handle.net/1843/35463
dc.language.iso.fl_str_mv por
language por
dc.rights.driver.fl_str_mv http://creativecommons.org/licenses/by-nc-nd/3.0/pt/
info:eu-repo/semantics/openAccess
rights_invalid_str_mv http://creativecommons.org/licenses/by-nc-nd/3.0/pt/
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
dc.publisher.none.fl_str_mv Universidade Federal de Minas Gerais
Brasil
ICB - DEPARTAMENTO DE BIOQUÍMICA E IMUNOLOGIA
Programa de Pós-Graduação em Bioquímica e Imunologia
UFMG
publisher.none.fl_str_mv Universidade Federal de Minas Gerais
Brasil
ICB - DEPARTAMENTO DE BIOQUÍMICA E IMUNOLOGIA
Programa de Pós-Graduação em Bioquímica e Imunologia
UFMG
dc.source.none.fl_str_mv reponame:Repositório Institucional da UFMG
instname:Universidade Federal de Minas Gerais (UFMG)
instacron:UFMG
instname_str Universidade Federal de Minas Gerais (UFMG)
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institution UFMG
reponame_str Repositório Institucional da UFMG
collection Repositório Institucional da UFMG
repository.name.fl_str_mv Repositório Institucional da UFMG - Universidade Federal de Minas Gerais (UFMG)
repository.mail.fl_str_mv repositorio@ufmg.br
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