Efeito da peçonha de Bothrops moojeni sobre formas promastigotas de Leishmania spp
Autor(a) principal: | |
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Data de Publicação: | 2008 |
Tipo de documento: | Dissertação |
Idioma: | por |
Título da fonte: | Repositório Institucional da UFU |
Texto Completo: | https://repositorio.ufu.br/handle/123456789/16632 |
Resumo: | In order to develop alternative tools to subdue infectious diseases, several substances have been tested toward treatment of parasitoses, such as Leishmaniasis. For this purpose, snake venoms have been used as future drugs that disrupt the viability of protozoan. In this report, we evaluated the effect of Bothrops moojeni venom and its fractions on viability, protein profile, infectivity and nitric oxide production of Leishmania promastigotes in vitro . Bothrops moojeni venom was fractionated into five protein fractions (E1 to E5) by ion exclusion chromatography and used to treat on L. (L.) amazonensis and L. (V.) braziliensis promastigote forms. The viability of Leishmania promastigotes was evaluated by MTT test and nitric oxide production was detected in supernatants of Leishmania culture. It was observed that E5 fraction strongly inhibited NO production in different concentrations in L. amazonensis, while in L. brasiliensis the NO production was enhance in ever concentration. It was also observed that crude venom of B. moojeni inhibited the viability of the both parasites in a dose dependent manner. However the E5 fraction only inhibited the viability of L. amazonensis. Also a peptide of 64 kDa of L (L.) amazonensis was apparently degraded in other two peptides with 56 and 51 kDa respectively. Moreover, when Leishmania spp was treated with E5 fraction and amphotericin B was not observed degradation of proteins. And after studies with the E5 fraction in DEAE (-) Sephacel, it was realized the exclusion cromatography and was obtained six new fractions (E5G1 and E5G6) and the cellular viability Leishmania (L.) amazonensis was sensitive to E5G1 (≥ 0.156 μg) and E5G5 (≥ 0.313μg). Adicionaly, it was analyzed the effect of Bothrops moojeni venon and the E5 fraction in the lesion progression of BALB/c mice infected with L. amazonensis and during the following period of lesion evolution (six weeks) it was verified that the thickness of the footpads of all infected animals with pre treated parasites did not increase in comparison to the non infected contralateral footpad. Thus, we demonstrated that Bothrops moojeni venom inhibited NO production, altered the protein profile and reduced the viability and infectivity of Leishmania promastigotes in vitro . |
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Efeito da peçonha de Bothrops moojeni sobre formas promastigotas de Leishmania sppLeishmania (Leishmania) amazonensisLeishmania (Viannia) braziliensisB. moojeniÓxido nítricoViabilidade de promastigotasLeishmanioseNitric oxidePromastigote viabilityCNPQ::CIENCIAS BIOLOGICAS::IMUNOLOGIA::IMUNOLOGIA APLICADAIn order to develop alternative tools to subdue infectious diseases, several substances have been tested toward treatment of parasitoses, such as Leishmaniasis. For this purpose, snake venoms have been used as future drugs that disrupt the viability of protozoan. In this report, we evaluated the effect of Bothrops moojeni venom and its fractions on viability, protein profile, infectivity and nitric oxide production of Leishmania promastigotes in vitro . Bothrops moojeni venom was fractionated into five protein fractions (E1 to E5) by ion exclusion chromatography and used to treat on L. (L.) amazonensis and L. (V.) braziliensis promastigote forms. The viability of Leishmania promastigotes was evaluated by MTT test and nitric oxide production was detected in supernatants of Leishmania culture. It was observed that E5 fraction strongly inhibited NO production in different concentrations in L. amazonensis, while in L. brasiliensis the NO production was enhance in ever concentration. It was also observed that crude venom of B. moojeni inhibited the viability of the both parasites in a dose dependent manner. However the E5 fraction only inhibited the viability of L. amazonensis. Also a peptide of 64 kDa of L (L.) amazonensis was apparently degraded in other two peptides with 56 and 51 kDa respectively. Moreover, when Leishmania spp was treated with E5 fraction and amphotericin B was not observed degradation of proteins. And after studies with the E5 fraction in DEAE (-) Sephacel, it was realized the exclusion cromatography and was obtained six new fractions (E5G1 and E5G6) and the cellular viability Leishmania (L.) amazonensis was sensitive to E5G1 (≥ 0.156 μg) and E5G5 (≥ 0.313μg). Adicionaly, it was analyzed the effect of Bothrops moojeni venon and the E5 fraction in the lesion progression of BALB/c mice infected with L. amazonensis and during the following period of lesion evolution (six weeks) it was verified that the thickness of the footpads of all infected animals with pre treated parasites did not increase in comparison to the non infected contralateral footpad. Thus, we demonstrated that Bothrops moojeni venom inhibited NO production, altered the protein profile and reduced the viability and infectivity of Leishmania promastigotes in vitro .Conselho Nacional de Desenvolvimento Científico e TecnológicoMestre em Imunologia e Parasitologia AplicadasO estudo do tratamento de doenças infecciosas vem desenvolvendo ferramentas alternativas e muitas substâncias estão sendo testadas no tratamento de parasitoses, como Leishmanioses. Com esta proposta, a peçonha de serpentes tem sido estudada como droga capaz de alterar a viabilidade de parasitos. Neste trabalho, foi avaliado o efeito da peçonha de Bothrops moojeni e suas Frações protéicas na viabilidade, na produção de NO, na infectividade e nas proteínas de promastigotas de Leishmania in vitro . A peçonha de Bothrops moojeni foi fracionada em cinco Frações (E1 a E5) por cromatografia de troca iônica e usadas no tratamento de promastigotas de L. (L.) amazonensis e L. (V.) braziliensis. A viabilidade foi analisada utilizando-se o MTT como substrato e a produção de óxido nítrico foi determinada no sobrenadante das culturas dos parasitos pelo método de Griess. Foi observado que a Fração E5 inibe fortemente a produção de NO em diferentes concentrações em L. amazonensis, enquanto em L. braziliensis a produção de NO apresentou aumento. Também foi observado que a peçonha inibe a viabilidade de ambos parasitos de forma dose-dependente, enquanto a Fração E5 inibe a viabilidade de L. amazonensis. Além disso, os parasitos tratados com peçonha apresentaram a proteína de 64 kDa de L (L.) amazonensis aparentemente degradada em duas outras proteínas de 56 e 51 kDa respectivamente. Quando Leishmania spp foi tratada com Fração E5 e com anfotericina B não foi observado nenhuma alteração no perfil eletroforético dos parasitos. E após os estudos com a Fração E5 obtida na cromatografia de troca iônica, a mesma foi eluída em cromatografia de exclusão molecular e foram obtidos seis novos picos de Frações protéicas (E5G1 a E5G6). Estes foram testados na viabilidade celular de Leishmania (L.) amazonensis e o parasito somente foi sensível às subFrações E5G1 (≥ 0.156 μg) e E5G5 (≥ 0.313μg). Adicionalmente foram analisados o efeito da peçonha de Bothrops moojeni e da Fração E5 na progressão da lesão de camundongos BALB/c infectados por L. amazonensis. Durante o período de acompanhamento da evolução da lesão (6 semanas), verificou-se que a espessura das patas dos animais infectados com parasitos tratados até este período, não aumentou em relação à pata contralateral não infectada. Assim, demonstramos que peçonha de Bothrops moojeni e suas Frações protéicas alteram o perfil protéico e reduzem a viabilidade celular e a produção de NO por Leishmania, levando a diminuição da infectividade dos parasitos tratados, observados pela ausência de lesão nas patas dos camundongos inoculados com parasitos tratados com as respectivas drogas.Universidade Federal de UberlândiaBRPrograma de Pós-graduação em Imunologia e Parasitologia AplicadasCiências BiológicasUFUOliveira, Fábio dehttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4799971D5Souza, Maria Aparecida dehttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4762379E9Abramo, Claricehttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4721158J4Castilhos, Patrícia de2016-06-22T18:46:32Z2009-03-172016-06-22T18:46:32Z2008-07-31info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisapplication/pdfapplication/pdfCASTILHOS, Patrícia de. Efeito da peçonha de Bothrops moojeni sobre formas promastigotas de Leishmania spp. 2008. 72 f. Dissertação (Mestrado em Ciências Biológicas) - Universidade Federal de Uberlândia, Uberlândia, 2008.https://repositorio.ufu.br/handle/123456789/16632porinfo:eu-repo/semantics/openAccessreponame:Repositório Institucional da UFUinstname:Universidade Federal de Uberlândia (UFU)instacron:UFU2016-06-23T07:37:21Zoai:repositorio.ufu.br:123456789/16632Repositório InstitucionalONGhttp://repositorio.ufu.br/oai/requestdiinf@dirbi.ufu.bropendoar:2016-06-23T07:37:21Repositório Institucional da UFU - Universidade Federal de Uberlândia (UFU)false |
dc.title.none.fl_str_mv |
Efeito da peçonha de Bothrops moojeni sobre formas promastigotas de Leishmania spp |
title |
Efeito da peçonha de Bothrops moojeni sobre formas promastigotas de Leishmania spp |
spellingShingle |
Efeito da peçonha de Bothrops moojeni sobre formas promastigotas de Leishmania spp Castilhos, Patrícia de Leishmania (Leishmania) amazonensis Leishmania (Viannia) braziliensis B. moojeni Óxido nítrico Viabilidade de promastigotas Leishmaniose Nitric oxide Promastigote viability CNPQ::CIENCIAS BIOLOGICAS::IMUNOLOGIA::IMUNOLOGIA APLICADA |
title_short |
Efeito da peçonha de Bothrops moojeni sobre formas promastigotas de Leishmania spp |
title_full |
Efeito da peçonha de Bothrops moojeni sobre formas promastigotas de Leishmania spp |
title_fullStr |
Efeito da peçonha de Bothrops moojeni sobre formas promastigotas de Leishmania spp |
title_full_unstemmed |
Efeito da peçonha de Bothrops moojeni sobre formas promastigotas de Leishmania spp |
title_sort |
Efeito da peçonha de Bothrops moojeni sobre formas promastigotas de Leishmania spp |
author |
Castilhos, Patrícia de |
author_facet |
Castilhos, Patrícia de |
author_role |
author |
dc.contributor.none.fl_str_mv |
Oliveira, Fábio de http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4799971D5 Souza, Maria Aparecida de http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4762379E9 Abramo, Clarice http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4721158J4 |
dc.contributor.author.fl_str_mv |
Castilhos, Patrícia de |
dc.subject.por.fl_str_mv |
Leishmania (Leishmania) amazonensis Leishmania (Viannia) braziliensis B. moojeni Óxido nítrico Viabilidade de promastigotas Leishmaniose Nitric oxide Promastigote viability CNPQ::CIENCIAS BIOLOGICAS::IMUNOLOGIA::IMUNOLOGIA APLICADA |
topic |
Leishmania (Leishmania) amazonensis Leishmania (Viannia) braziliensis B. moojeni Óxido nítrico Viabilidade de promastigotas Leishmaniose Nitric oxide Promastigote viability CNPQ::CIENCIAS BIOLOGICAS::IMUNOLOGIA::IMUNOLOGIA APLICADA |
description |
In order to develop alternative tools to subdue infectious diseases, several substances have been tested toward treatment of parasitoses, such as Leishmaniasis. For this purpose, snake venoms have been used as future drugs that disrupt the viability of protozoan. In this report, we evaluated the effect of Bothrops moojeni venom and its fractions on viability, protein profile, infectivity and nitric oxide production of Leishmania promastigotes in vitro . Bothrops moojeni venom was fractionated into five protein fractions (E1 to E5) by ion exclusion chromatography and used to treat on L. (L.) amazonensis and L. (V.) braziliensis promastigote forms. The viability of Leishmania promastigotes was evaluated by MTT test and nitric oxide production was detected in supernatants of Leishmania culture. It was observed that E5 fraction strongly inhibited NO production in different concentrations in L. amazonensis, while in L. brasiliensis the NO production was enhance in ever concentration. It was also observed that crude venom of B. moojeni inhibited the viability of the both parasites in a dose dependent manner. However the E5 fraction only inhibited the viability of L. amazonensis. Also a peptide of 64 kDa of L (L.) amazonensis was apparently degraded in other two peptides with 56 and 51 kDa respectively. Moreover, when Leishmania spp was treated with E5 fraction and amphotericin B was not observed degradation of proteins. And after studies with the E5 fraction in DEAE (-) Sephacel, it was realized the exclusion cromatography and was obtained six new fractions (E5G1 and E5G6) and the cellular viability Leishmania (L.) amazonensis was sensitive to E5G1 (≥ 0.156 μg) and E5G5 (≥ 0.313μg). Adicionaly, it was analyzed the effect of Bothrops moojeni venon and the E5 fraction in the lesion progression of BALB/c mice infected with L. amazonensis and during the following period of lesion evolution (six weeks) it was verified that the thickness of the footpads of all infected animals with pre treated parasites did not increase in comparison to the non infected contralateral footpad. Thus, we demonstrated that Bothrops moojeni venom inhibited NO production, altered the protein profile and reduced the viability and infectivity of Leishmania promastigotes in vitro . |
publishDate |
2008 |
dc.date.none.fl_str_mv |
2008-07-31 2009-03-17 2016-06-22T18:46:32Z 2016-06-22T18:46:32Z |
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 |
CASTILHOS, Patrícia de. Efeito da peçonha de Bothrops moojeni sobre formas promastigotas de Leishmania spp. 2008. 72 f. Dissertação (Mestrado em Ciências Biológicas) - Universidade Federal de Uberlândia, Uberlândia, 2008. https://repositorio.ufu.br/handle/123456789/16632 |
identifier_str_mv |
CASTILHOS, Patrícia de. Efeito da peçonha de Bothrops moojeni sobre formas promastigotas de Leishmania spp. 2008. 72 f. Dissertação (Mestrado em Ciências Biológicas) - Universidade Federal de Uberlândia, Uberlândia, 2008. |
url |
https://repositorio.ufu.br/handle/123456789/16632 |
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 Uberlândia BR Programa de Pós-graduação em Imunologia e Parasitologia Aplicadas Ciências Biológicas UFU |
publisher.none.fl_str_mv |
Universidade Federal de Uberlândia BR Programa de Pós-graduação em Imunologia e Parasitologia Aplicadas Ciências Biológicas UFU |
dc.source.none.fl_str_mv |
reponame:Repositório Institucional da UFU instname:Universidade Federal de Uberlândia (UFU) instacron:UFU |
instname_str |
Universidade Federal de Uberlândia (UFU) |
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UFU |
institution |
UFU |
reponame_str |
Repositório Institucional da UFU |
collection |
Repositório Institucional da UFU |
repository.name.fl_str_mv |
Repositório Institucional da UFU - Universidade Federal de Uberlândia (UFU) |
repository.mail.fl_str_mv |
diinf@dirbi.ufu.br |
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1805569642604789760 |