Avaliação citotóxica, mutagênica, carcinogênica e in silico do ácido betulínico
Autor(a) principal: | |
---|---|
Data de Publicação: | 2022 |
Tipo de documento: | Tese |
Idioma: | por |
Título da fonte: | Repositório Institucional da UFU |
Texto Completo: | https://repositorio.ufu.br/handle/123456789/35015 http://doi.org/10.14393/ufu.te.2022.5004 |
Resumo: | Betulinic acid (AB) is a pentacyclic triterpenoid found in several plant species. Here, we examine the genotoxicity and mutagenicity of BA alone or in combination with URE using the mouse bone marrow cell micronucleus assay and the somatic mutation and recombination test in Drosophila melanogaster. The findings revealed that the isolated AB was not genotoxic but reduced the frequency of micronuclei when compared to the positive control. No significant differences in cytotoxicity were observed. Biochemical analyzes did not show significant differences for hepatic (AST and ALT) or renal (creatinine and urea) function parameters, indicating the absence of hepatotoxic and nephrotoxic effects. AB alone did not increase the frequency of mutant spots but reduced the total frequency of mutant spots when co-administered with URE in both the ST and HB crosses. In addition, AB reduced the recombinogenic effect of URE at the highest concentrations of both crosses. We also evaluated AB alone for carcinogenicity and combined with DXR (0.4 mM) for anticarcinogenicity, using the Epithelial Tumor Test (ETT) in Drosophila melanogaster. AB alone did not significantly increase tumor frequency but was able to reduce overall tumor frequency when co-administered with DXR at all concentrations. MCF10A (non-tumorigenic) and MDA-MB-231 (tumorigenic) triple-negative mammary cells were treated with AB at different concentrations for 24 and 48 h and cell viability was evaluated by the MTT tetrazolium salt colorimetric assay. AB reduced the cell viability of MCF10A and MDA-MB-231 cells in a concentration-dependent manner compared to control (untreated cells). However, the cytotoxic effect of AB was more significant after 48 hours, where the reduction in growth viability was more efficient in MDA-MB-231 cells. The LDH release assay was used to measure the cell damage caused by AB. It was observed that there was no significant difference in LDH release when comparing untreated and AB-treated cells. To elucidate some of the ways in which AB works, we performed an in silico molecular docking analysis. The result of the search for the most likely targets of AB revealed that SUMO (SAE1/UBA2), DNA POLβ and AKR1B10 had the highest percentages of chance of being targets of the AB ligand. In conclusion, under experimental conditions, BA has modulatory effects on URE-induced genotoxicity in mice as well as in D. melanogaster somatic cells. our data suggest that BA can modulate the carcinogenic effect of DXR, inducing damaged cells to apoptosis through different pathways involving SUMO proteins (SAE1 and UBA2), DNA POLβ and AKR1B10. New studies must be carried out using other reference carcinogens and different organisms to confirm the use of this compound in the production of new drugs. |
id |
UFU_3c41542351ca96f1123c90073450ea7d |
---|---|
oai_identifier_str |
oai:repositorio.ufu.br:123456789/35015 |
network_acronym_str |
UFU |
network_name_str |
Repositório Institucional da UFU |
repository_id_str |
|
spelling |
Avaliação citotóxica, mutagênica, carcinogênica e in silico do ácido betulínicoCytotoxic, mutagenic, carcinogenic and in silico evaluation of betulin acidAntigenotoxicidadeAntimutagenicidadeAnticarcinogenicidadeCitocromo P450Teste de micronúcleoTeste de mutação e recombinação somáticaTeste de tumor epitelialEfeito apoptóticoDocking molecularCNPQ::CIENCIAS BIOLOGICAS::GENETICAGenéticaCarcinogêneseAgentes antineoplásicosÁcidos - DeposiçãoBetulinic acid (AB) is a pentacyclic triterpenoid found in several plant species. Here, we examine the genotoxicity and mutagenicity of BA alone or in combination with URE using the mouse bone marrow cell micronucleus assay and the somatic mutation and recombination test in Drosophila melanogaster. The findings revealed that the isolated AB was not genotoxic but reduced the frequency of micronuclei when compared to the positive control. No significant differences in cytotoxicity were observed. Biochemical analyzes did not show significant differences for hepatic (AST and ALT) or renal (creatinine and urea) function parameters, indicating the absence of hepatotoxic and nephrotoxic effects. AB alone did not increase the frequency of mutant spots but reduced the total frequency of mutant spots when co-administered with URE in both the ST and HB crosses. In addition, AB reduced the recombinogenic effect of URE at the highest concentrations of both crosses. We also evaluated AB alone for carcinogenicity and combined with DXR (0.4 mM) for anticarcinogenicity, using the Epithelial Tumor Test (ETT) in Drosophila melanogaster. AB alone did not significantly increase tumor frequency but was able to reduce overall tumor frequency when co-administered with DXR at all concentrations. MCF10A (non-tumorigenic) and MDA-MB-231 (tumorigenic) triple-negative mammary cells were treated with AB at different concentrations for 24 and 48 h and cell viability was evaluated by the MTT tetrazolium salt colorimetric assay. AB reduced the cell viability of MCF10A and MDA-MB-231 cells in a concentration-dependent manner compared to control (untreated cells). However, the cytotoxic effect of AB was more significant after 48 hours, where the reduction in growth viability was more efficient in MDA-MB-231 cells. The LDH release assay was used to measure the cell damage caused by AB. It was observed that there was no significant difference in LDH release when comparing untreated and AB-treated cells. To elucidate some of the ways in which AB works, we performed an in silico molecular docking analysis. The result of the search for the most likely targets of AB revealed that SUMO (SAE1/UBA2), DNA POLβ and AKR1B10 had the highest percentages of chance of being targets of the AB ligand. In conclusion, under experimental conditions, BA has modulatory effects on URE-induced genotoxicity in mice as well as in D. melanogaster somatic cells. our data suggest that BA can modulate the carcinogenic effect of DXR, inducing damaged cells to apoptosis through different pathways involving SUMO proteins (SAE1 and UBA2), DNA POLβ and AKR1B10. New studies must be carried out using other reference carcinogens and different organisms to confirm the use of this compound in the production of new drugs.CAPES - Coordenação de Aperfeiçoamento de Pessoal de Nível SuperiorTese (Doutorado)O ácido betulínico (AB) é um triterpenóide pentacíclico encontrado em várias espécies de plantas. Aqui, examinamos a genotoxicidade e mutagenicidade de BA sozinho ou em combinação com URE usando o ensaio de micronúcleo em células de medula óssea de camundongos e o teste de mutação e recombinação somática em Drosophila melanogaster. Os achados revelaram que o AB isolado não foi genotóxico, mas reduziu a frequência de micronúcleos quando comparado ao controle positivo. Não foram observadas diferenças significativas na citotoxicidade. As análises bioquímicas não mostraram diferenças significativas para os parâmetros de função hepática (AST e ALT) ou renal (creatinina e ureia), indicando ausência de efeitos hepatotóxicos e nefrotóxicos. O AB isolado não aumentou a frequência de manchas mutantes, mas reduziu a frequência total de manchas mutantes quando coadministrado com URE em ambos os cruzamentos ST e HB. Além disso, o AB reduziu o efeito recombinogênico da URE nas concentrações mais altas de ambos os cruzamentos. Nós também avaliamos o AB isolado para carcinogenicidade e combinado com DXR (0,4 mM) para anticarcinogenicidade, usando o Epithelial Tumor Test (ETT) em Drosophila melanogaster. O AB isolado não aumentou significativamente a frequência de tumores, mas foi capaz de reduzir a frequência total de tumores quando coadministrado com DXR, em todas as concentrações. Células mamárias triplo-negativas MCF10A (não tumorigênicas) e MDA-MB-231 (tumorigênicas) foram tratadas com AB em diferentes concentrações por 24 e 48 h e a viabilidade celular foi avaliada pelo ensaio colorimétrico de sal de tetrazólio MTT. AB reduziu a viabilidade celular de células MCF10A e MDA-MB-231 de uma maneira dependente da concentração em comparação com o controle (células não tratadas). No entanto, o efeito citotóxico do AB foi mais significativo após 48 horas, onde a redução da viabilidade de crescimento foi mais eficiente nas células MDA-MB-231. O ensaio de liberação de LDH foi usado para medir o dano celular causado por AB. Observou-se que não houve diferença significativa na liberação de LDH ao comparar células não tratadas e tratadas com AB. Para elucidar algumas das formas de atuação do AB, realizamos uma análise in silico de docking molecular. O resultado da busca pelos alvos mais prováveis do AB revelou que SUMO (SAE1/UBA2), DNA POLβ e AKR1B10 tiveram os maiores percentuais de chance de serem alvos do ligante AB. Em conclusão, em condições experimentais, o BA tem efeitos moduladores sobre a genotoxicidade induzida por URE em camundongos, bem como em células somáticas de D. melanogaster. nossos dados sugerem que o BA pode modular o efeito carcinogênico do DXR, induzindo células danificadas à apoptose através de diferentes vias envolvendo proteínas SUMO (SAE1 e UBA2), DNA POLβ e AKR1B10. Novos estudos devem ser realizados utilizando outros carcinógenos de referência e diferentes organismos para confirmar o uso deste composto na produção de novos medicamentos.2024-04-19Universidade Federal de UberlândiaBrasilPrograma de Pós-graduação em Genética e BioquímicaSpano, Mario Antoniohttp://lattes.cnpq.br/1910653028916454Rezende, Alexandre Azenha Alves dehttp://lattes.cnpq.br/9751160400590652Fragiorge, Edson Joséhttp://lattes.cnpq.br/8877087834110730Chen, Lee Chenhttp://lattes.cnpq.br/4621907105842007Morelli, Sandrahttp://lattes.cnpq.br/1661483102212717Oliveira, Victor Constante2022-05-09T18:07:23Z2022-05-09T18:07:23Z2022-01-28info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/doctoralThesisapplication/pdfOLIVEIRA, Victor Constante. Avaliação citotóxica, mutagênica, carcinogênica e in silico do ácido betulínico. 2022. 140 f. Tese (Doutorado em Genética e Bioquímica) - Universidade Federal de Uberlândia, Uberlândia, 2022. DOI http://doi.org/10.14393/ufu.te.2022.5004.https://repositorio.ufu.br/handle/123456789/35015http://doi.org/10.14393/ufu.te.2022.5004porhttp://creativecommons.org/licenses/by-nc-nd/3.0/us/info:eu-repo/semantics/embargoedAccessreponame:Repositório Institucional da UFUinstname:Universidade Federal de Uberlândia (UFU)instacron:UFU2024-02-08T19:35:47Zoai:repositorio.ufu.br:123456789/35015Repositório InstitucionalONGhttp://repositorio.ufu.br/oai/requestdiinf@dirbi.ufu.bropendoar:2024-02-08T19:35:47Repositório Institucional da UFU - Universidade Federal de Uberlândia (UFU)false |
dc.title.none.fl_str_mv |
Avaliação citotóxica, mutagênica, carcinogênica e in silico do ácido betulínico Cytotoxic, mutagenic, carcinogenic and in silico evaluation of betulin acid |
title |
Avaliação citotóxica, mutagênica, carcinogênica e in silico do ácido betulínico |
spellingShingle |
Avaliação citotóxica, mutagênica, carcinogênica e in silico do ácido betulínico Oliveira, Victor Constante Antigenotoxicidade Antimutagenicidade Anticarcinogenicidade Citocromo P450 Teste de micronúcleo Teste de mutação e recombinação somática Teste de tumor epitelial Efeito apoptótico Docking molecular CNPQ::CIENCIAS BIOLOGICAS::GENETICA Genética Carcinogênese Agentes antineoplásicos Ácidos - Deposição |
title_short |
Avaliação citotóxica, mutagênica, carcinogênica e in silico do ácido betulínico |
title_full |
Avaliação citotóxica, mutagênica, carcinogênica e in silico do ácido betulínico |
title_fullStr |
Avaliação citotóxica, mutagênica, carcinogênica e in silico do ácido betulínico |
title_full_unstemmed |
Avaliação citotóxica, mutagênica, carcinogênica e in silico do ácido betulínico |
title_sort |
Avaliação citotóxica, mutagênica, carcinogênica e in silico do ácido betulínico |
author |
Oliveira, Victor Constante |
author_facet |
Oliveira, Victor Constante |
author_role |
author |
dc.contributor.none.fl_str_mv |
Spano, Mario Antonio http://lattes.cnpq.br/1910653028916454 Rezende, Alexandre Azenha Alves de http://lattes.cnpq.br/9751160400590652 Fragiorge, Edson José http://lattes.cnpq.br/8877087834110730 Chen, Lee Chen http://lattes.cnpq.br/4621907105842007 Morelli, Sandra http://lattes.cnpq.br/1661483102212717 |
dc.contributor.author.fl_str_mv |
Oliveira, Victor Constante |
dc.subject.por.fl_str_mv |
Antigenotoxicidade Antimutagenicidade Anticarcinogenicidade Citocromo P450 Teste de micronúcleo Teste de mutação e recombinação somática Teste de tumor epitelial Efeito apoptótico Docking molecular CNPQ::CIENCIAS BIOLOGICAS::GENETICA Genética Carcinogênese Agentes antineoplásicos Ácidos - Deposição |
topic |
Antigenotoxicidade Antimutagenicidade Anticarcinogenicidade Citocromo P450 Teste de micronúcleo Teste de mutação e recombinação somática Teste de tumor epitelial Efeito apoptótico Docking molecular CNPQ::CIENCIAS BIOLOGICAS::GENETICA Genética Carcinogênese Agentes antineoplásicos Ácidos - Deposição |
description |
Betulinic acid (AB) is a pentacyclic triterpenoid found in several plant species. Here, we examine the genotoxicity and mutagenicity of BA alone or in combination with URE using the mouse bone marrow cell micronucleus assay and the somatic mutation and recombination test in Drosophila melanogaster. The findings revealed that the isolated AB was not genotoxic but reduced the frequency of micronuclei when compared to the positive control. No significant differences in cytotoxicity were observed. Biochemical analyzes did not show significant differences for hepatic (AST and ALT) or renal (creatinine and urea) function parameters, indicating the absence of hepatotoxic and nephrotoxic effects. AB alone did not increase the frequency of mutant spots but reduced the total frequency of mutant spots when co-administered with URE in both the ST and HB crosses. In addition, AB reduced the recombinogenic effect of URE at the highest concentrations of both crosses. We also evaluated AB alone for carcinogenicity and combined with DXR (0.4 mM) for anticarcinogenicity, using the Epithelial Tumor Test (ETT) in Drosophila melanogaster. AB alone did not significantly increase tumor frequency but was able to reduce overall tumor frequency when co-administered with DXR at all concentrations. MCF10A (non-tumorigenic) and MDA-MB-231 (tumorigenic) triple-negative mammary cells were treated with AB at different concentrations for 24 and 48 h and cell viability was evaluated by the MTT tetrazolium salt colorimetric assay. AB reduced the cell viability of MCF10A and MDA-MB-231 cells in a concentration-dependent manner compared to control (untreated cells). However, the cytotoxic effect of AB was more significant after 48 hours, where the reduction in growth viability was more efficient in MDA-MB-231 cells. The LDH release assay was used to measure the cell damage caused by AB. It was observed that there was no significant difference in LDH release when comparing untreated and AB-treated cells. To elucidate some of the ways in which AB works, we performed an in silico molecular docking analysis. The result of the search for the most likely targets of AB revealed that SUMO (SAE1/UBA2), DNA POLβ and AKR1B10 had the highest percentages of chance of being targets of the AB ligand. In conclusion, under experimental conditions, BA has modulatory effects on URE-induced genotoxicity in mice as well as in D. melanogaster somatic cells. our data suggest that BA can modulate the carcinogenic effect of DXR, inducing damaged cells to apoptosis through different pathways involving SUMO proteins (SAE1 and UBA2), DNA POLβ and AKR1B10. New studies must be carried out using other reference carcinogens and different organisms to confirm the use of this compound in the production of new drugs. |
publishDate |
2022 |
dc.date.none.fl_str_mv |
2022-05-09T18:07:23Z 2022-05-09T18:07:23Z 2022-01-28 |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/doctoralThesis |
format |
doctoralThesis |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
OLIVEIRA, Victor Constante. Avaliação citotóxica, mutagênica, carcinogênica e in silico do ácido betulínico. 2022. 140 f. Tese (Doutorado em Genética e Bioquímica) - Universidade Federal de Uberlândia, Uberlândia, 2022. DOI http://doi.org/10.14393/ufu.te.2022.5004. https://repositorio.ufu.br/handle/123456789/35015 http://doi.org/10.14393/ufu.te.2022.5004 |
identifier_str_mv |
OLIVEIRA, Victor Constante. Avaliação citotóxica, mutagênica, carcinogênica e in silico do ácido betulínico. 2022. 140 f. Tese (Doutorado em Genética e Bioquímica) - Universidade Federal de Uberlândia, Uberlândia, 2022. DOI http://doi.org/10.14393/ufu.te.2022.5004. |
url |
https://repositorio.ufu.br/handle/123456789/35015 http://doi.org/10.14393/ufu.te.2022.5004 |
dc.language.iso.fl_str_mv |
por |
language |
por |
dc.rights.driver.fl_str_mv |
http://creativecommons.org/licenses/by-nc-nd/3.0/us/ info:eu-repo/semantics/embargoedAccess |
rights_invalid_str_mv |
http://creativecommons.org/licenses/by-nc-nd/3.0/us/ |
eu_rights_str_mv |
embargoedAccess |
dc.format.none.fl_str_mv |
application/pdf |
dc.publisher.none.fl_str_mv |
Universidade Federal de Uberlândia Brasil Programa de Pós-graduação em Genética e Bioquímica |
publisher.none.fl_str_mv |
Universidade Federal de Uberlândia Brasil Programa de Pós-graduação em Genética e Bioquímica |
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) |
instacron_str |
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 |
_version_ |
1805569660311044096 |