Avaliação da citotoxicidade e da atividade antimicrobiana e antioxidante do extrato hidroetanólico de leonotis nepetaefolia (l) r. Br.
Autor(a) principal: | |
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Data de Publicação: | 2013 |
Tipo de documento: | Dissertação |
Idioma: | por |
Título da fonte: | Repositório Institucional da UFMT |
Texto Completo: | http://ri.ufmt.br/handle/1/1590 |
Resumo: | Leonotis nepetifolia (L.) R. Br (Lamiaceae), popularly known as “cordão-de- frade”, is a sub-spontaneous shrub. The tea from the whole plant is used in the treatment of infections, inflammations, wounds and stomach ulcers. The present study evaluates the potential antimicrobial, antioxidant and cytotoxicity of the hydroethanolic extract of L. nepetifolia (HELn). The whole plant was washed, dried, powdered and the resulting powder was macerated in hydroethanolic solution (1:5, w/v), filtered, concentrated in rotaryevaporator and the residual solvent was eliminated in hotair oven, thus obtaining HELn. Chinese hamster ovary cells (CHO-K1) were used for cytotoxicity studies by Alamar blue method. To evaluate the antimicrobial activity against a panel of Gram-positive and Gram-negative bacteria and yeasts and filamentos fungi, disk diffusion test and broth microdilution were used. Drugs with inhibition zones of ≥ 10 mm and minimal inhibitory concentration (MIC) <100 µg/mL were considered very active. To characterize the nature of the antibacterial activity, antibiosis mechanism study was conducted, evaluating the growth of sensitive bacteria to concentrations of HELn ≥ MIC within 24 h. To assess the antibacterial mode of action of HELn on the permeability of the outer membrane, synergism between the extract (0.5 MIC) and hydrophobic antibiotics- erythromycin and rifampicin (0.68 to 20 µg/mL), was determined in Shigella flexneri. However, the mode of action of of HELn on the cytoplasmic membrane was evaluated by measuring the potassium efflux (2 MIC) and nucleotides leakages (1 MIC), using electrolytes analyzer and spectrophotometer respectively, in strains of E. faecalis and S. flexneri. The antioxidant activity was determined in vitro by DPPH, NO and H2O2 radical scavenging assays. The IC50 of HELn in the cytotoxicity assay was greater than 200 µg/mL. HELn demostrated a good spectrum of antibacterial activity in broth microdilution assay against S. flexneri, S. aureus and E. faecalis (MIC= 6.25 µg/mL), H. pylori (MIC= 25 µg/mL) and S. pyogenes (MIC= 50 µg/mL). With chloramphenicol, the MIC varied between 3.12 µg/mL (S. flexneri ) and 100 µg/mL (P. aeruginosa). In the disk diffusion assay, HELn was inactive, while chloramphenicol produced inhibition halos of 30 mm against all strains tested. HELn presented a bacteriostatic action against the sensitive bacterial strains. In the presence of HELn, S. flexneri became more sensitive to the probe antibiotics at MIC of erythromycin (1 µg/mL) and below the MIC of rifampicin (5 µg/mL). HELn caused an intense damage to the cytoplasmic membrane, reaching a maximum K+ efflux of 11.7 ± 0.6 ppm and 12.87 ± 0.3 ppm, and of the nucleotide 22% (6th hour) and 46% (12th time ) in E. faecalis and S. flexneri, respectively. EHLn had no effect on fungal growth, unlike amphotericin B, which was active in both models of disk diffusion and broth microdilution, attaining maxima activity against yeasts (35 mm) and filamentous fungi (3.12 µg/mL). HELn demonstrated weak antioxidant activity with IC50 (µg/mL) of 128.94 ± 8.7, 190.93 ± 0.01 and > 250 for DPPH, NO and H2O2 radical scavenging assays respectively. The results indicate that HELn has low cytotoxicity and antioxidant activity and potent antibacterial activity, especially against Gram-positive bacteria,. It is bacteriostatic in nature, possibly acting at the level of the cell wall and cytoplasmic membrane thus supporting its popular use in infectious processes. In addition, the presence of phenols, flavonoides, carotenoids, fatty acids and esteroids, described in the literature to posses antioxidant and antimicrobial, were detected in the composition of HELn |
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Avaliação da citotoxicidade e da atividade antimicrobiana e antioxidante do extrato hidroetanólico de leonotis nepetaefolia (l) r. Br.Leonotis nepetefoliaAntimicrobianoCitotoxicidadeAntioxidanteCNPQ::CIENCIAS DA SAUDE::MEDICINALeonotis nepetefoliaAntimicrobialCytotoxicityAntioxidantLeonotis nepetifolia (L.) R. Br (Lamiaceae), popularly known as “cordão-de- frade”, is a sub-spontaneous shrub. The tea from the whole plant is used in the treatment of infections, inflammations, wounds and stomach ulcers. The present study evaluates the potential antimicrobial, antioxidant and cytotoxicity of the hydroethanolic extract of L. nepetifolia (HELn). The whole plant was washed, dried, powdered and the resulting powder was macerated in hydroethanolic solution (1:5, w/v), filtered, concentrated in rotaryevaporator and the residual solvent was eliminated in hotair oven, thus obtaining HELn. Chinese hamster ovary cells (CHO-K1) were used for cytotoxicity studies by Alamar blue method. To evaluate the antimicrobial activity against a panel of Gram-positive and Gram-negative bacteria and yeasts and filamentos fungi, disk diffusion test and broth microdilution were used. Drugs with inhibition zones of ≥ 10 mm and minimal inhibitory concentration (MIC) <100 µg/mL were considered very active. To characterize the nature of the antibacterial activity, antibiosis mechanism study was conducted, evaluating the growth of sensitive bacteria to concentrations of HELn ≥ MIC within 24 h. To assess the antibacterial mode of action of HELn on the permeability of the outer membrane, synergism between the extract (0.5 MIC) and hydrophobic antibiotics- erythromycin and rifampicin (0.68 to 20 µg/mL), was determined in Shigella flexneri. However, the mode of action of of HELn on the cytoplasmic membrane was evaluated by measuring the potassium efflux (2 MIC) and nucleotides leakages (1 MIC), using electrolytes analyzer and spectrophotometer respectively, in strains of E. faecalis and S. flexneri. The antioxidant activity was determined in vitro by DPPH, NO and H2O2 radical scavenging assays. The IC50 of HELn in the cytotoxicity assay was greater than 200 µg/mL. HELn demostrated a good spectrum of antibacterial activity in broth microdilution assay against S. flexneri, S. aureus and E. faecalis (MIC= 6.25 µg/mL), H. pylori (MIC= 25 µg/mL) and S. pyogenes (MIC= 50 µg/mL). With chloramphenicol, the MIC varied between 3.12 µg/mL (S. flexneri ) and 100 µg/mL (P. aeruginosa). In the disk diffusion assay, HELn was inactive, while chloramphenicol produced inhibition halos of 30 mm against all strains tested. HELn presented a bacteriostatic action against the sensitive bacterial strains. In the presence of HELn, S. flexneri became more sensitive to the probe antibiotics at MIC of erythromycin (1 µg/mL) and below the MIC of rifampicin (5 µg/mL). HELn caused an intense damage to the cytoplasmic membrane, reaching a maximum K+ efflux of 11.7 ± 0.6 ppm and 12.87 ± 0.3 ppm, and of the nucleotide 22% (6th hour) and 46% (12th time ) in E. faecalis and S. flexneri, respectively. EHLn had no effect on fungal growth, unlike amphotericin B, which was active in both models of disk diffusion and broth microdilution, attaining maxima activity against yeasts (35 mm) and filamentous fungi (3.12 µg/mL). HELn demonstrated weak antioxidant activity with IC50 (µg/mL) of 128.94 ± 8.7, 190.93 ± 0.01 and > 250 for DPPH, NO and H2O2 radical scavenging assays respectively. The results indicate that HELn has low cytotoxicity and antioxidant activity and potent antibacterial activity, especially against Gram-positive bacteria,. It is bacteriostatic in nature, possibly acting at the level of the cell wall and cytoplasmic membrane thus supporting its popular use in infectious processes. In addition, the presence of phenols, flavonoides, carotenoids, fatty acids and esteroids, described in the literature to posses antioxidant and antimicrobial, were detected in the composition of HELnCNPqLeonotis nepetifolia (L.) R. Br. (Lamiaceae), popularmente conhecida como cordão-de-frade é um arbusto subespontâneo, cujo chá da planta inteira é usado popularmente na Amazônia Legal para infecções, inflamações, feridas e úlceras gástricas. O presente estudo avalia o potencial antimicrobiano, antioxidante e citotóxico do extrato hidroetanólico de L. nepetifolia (EHLn). A planta inteira foi limpa, seca, triturada e o pó resultante macerado em soulção hiroetanólica (1:5, p/v), filtrado, concentrado em rotaevaporador e o solvente residual eliminado em estufa, obtendo-se assim o EHLn. Células de ovário de hamster chinês (CHO-k1) foram utilizadas para a avaliação da citotoxicidade, pelo método de “Alamar blue”. Para avaliação da atividade antimicrobiana, frente a um painel de bactérias Gram-positivas e Gram-negativas e fungos leveduriformes e fialamentosos, foram utilizados os ensaios de difusão em disco e microdiluição em caldo, consdirerando-se muito ativas drogas com halos de inibição ≥ 10 mm e concentração inibitória mínima (CIM) < 100 µg/mL. Para caracterização da natureza da atividade antibacteriana, foi realizado estudo do mecanismo de antibiose, avaliando-se o crescimento de bactérias sensíveis a concentrações do EHLn ≥ CIM em até 24 h. Para avaliação do modo de ação antibacteriano do EHLn sobre a permeabilidade de membrana externa, foi determinado o sinergismo entre o extrato (0,5 CIM) e os antibióticos hidrofóbicos eritromicina e rifampicina (0,68 a 20 µg/mL), em cepas de Shigella flexneri. Já o modo de ação do EHLn sobre a membrana citoplasmática foi avaliado pela medida do efluxo de potássio (2 CIM) e nucleotídeos (1 CIM), utlizando-se analisador de eletrólitos e espectrofotômetro, respectivamente, em cepas de E. faecalis e S. flexneri. A atividade antioxidante foi determinada pelo ensaio da captura dos radicais DPPH, NO e H2O2 in vitro. No ensaio de citotoxicidade, o EHLn apresentou CI50 > 200 µg/mL. O EHLn apresentou um bom espectro de ação antibacteriana, em ensaio de microdiluição em caldo, frente à S. flexneri, S. aureus e E. faecalis (CIM= 6,25 µg/mL), H. pylori (CIM= 25 µg/mL) e S. pyogenes (CIM= 50 µg/mL). Com cloranfenicol, a CIM bacteriana variou de 3,12 µg/mL (S. flexneri ) a 100 µg/mL (P. aeruginosa). No ensaio de difusão em disco, o EHLn mostrou-se inativo, enquanto o cloranfenicol produziu halos de inibição de 30 mm para todas as cepas testadas. O EHLn apresentou ação bacteriostática frente às cepas sensíveis. Na presença do EHLn, a S. flexneri tornou-se mais sensível aos antibióticos-prova, na CIM da eritromicina (1 µg/mL) e abaixo da CIM da rifampicina (5 µg/mL). O EHLn causou intenso dano da membrana citoplasmática, atingindo um efluxo máximo de K + de 11,7±0,6 ppm e 12,87±0,3 ppm, e de nucleotídeos de 22% (6ª hora) e 46% (12ª hora) , para E. faecalis e S. flexneri, respectivamente. O EHLn não interferiu no crescimento fúngico, ao contrário da anfotericina B , que foi ativa em ambos os modelos de difusão em disco e microdiluição em caldo, atingindo maior atividade frente a leveduras (35 mm) e filamentosos ( 3,12 µg/mL). O EHLn apresentou fraca atividade antioxidante, com CI50 (μg/mL) de 128,94±8,7, 190,93 ± 0,01 e >250, para as capturas dos radicais DPPH, NO e H2O2, respectivamente. Os resultados indicam que o EHLn apresenta baixa citotoxidade e atividade antioxidante e potente atividade antibacteriana, especialmente frente a bactérias Gram-positivas, de natureza bacteriostática, possivelmente agindo a nível da parede celular e membrana citoplasmática, respaldando seu uso popular em processos infeciosos. Além do mais, apresenta em sua composição fenois, flavonóides, carotenoides, ácidos graxos e esteroides, descritos como antioxidantes e antimicrobianos.Universidade Federal de Mato GrossoBrasilFaculdade de Medicina (FM)UFMT CUC - CuiabáPrograma de Pós-Graduação em Ciências da SaúdeMartins, Domingos Tabajara de Oliveirahttp://lattes.cnpq.br/3794477872946546Martins, Domingos Tabajara de Oliveira109.726.923-04http://lattes.cnpq.br/3794477872946546Slhessarenko, Renata Dezengrini006.990.439-13http://lattes.cnpq.br/3790088995387532109.726.923-04Cruz, Alexandre Bella480.657.909-25http://lattes.cnpq.br/1596661935560930Lopes, Lousa879.675.298-04http://lattes.cnpq.br/8843725522665416Oliveira, Darley Maria2019-11-21T13:29:55Z2013-06-052019-11-21T13:29:55Z2013-05-23info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisOLIVEIRA, Darley Maria. Avaliação da citotoxicidade e da atividade antimicrobiana e antioxidante do extrato hidroetanólico de leonotis nepetaefolia (l) r. Br. 2013. 96 f. Dissertação (Mestrado em Ciências da Saúde) - Universidade Federal de Mato Grosso, Faculdade de Medicina, Cuiabá, 2013.http://ri.ufmt.br/handle/1/1590porinfo:eu-repo/semantics/openAccessreponame:Repositório Institucional da UFMTinstname:Universidade Federal de Mato Grosso (UFMT)instacron:UFMT2019-11-24T06:06:33Zoai:localhost:1/1590Repositório InstitucionalPUBhttp://ri.ufmt.br/oai/requestjordanbiblio@gmail.comopendoar:2019-11-24T06:06:33Repositório Institucional da UFMT - Universidade Federal de Mato Grosso (UFMT)false |
dc.title.none.fl_str_mv |
Avaliação da citotoxicidade e da atividade antimicrobiana e antioxidante do extrato hidroetanólico de leonotis nepetaefolia (l) r. Br. |
title |
Avaliação da citotoxicidade e da atividade antimicrobiana e antioxidante do extrato hidroetanólico de leonotis nepetaefolia (l) r. Br. |
spellingShingle |
Avaliação da citotoxicidade e da atividade antimicrobiana e antioxidante do extrato hidroetanólico de leonotis nepetaefolia (l) r. Br. Oliveira, Darley Maria Leonotis nepetefolia Antimicrobiano Citotoxicidade Antioxidante CNPQ::CIENCIAS DA SAUDE::MEDICINA Leonotis nepetefolia Antimicrobial Cytotoxicity Antioxidant |
title_short |
Avaliação da citotoxicidade e da atividade antimicrobiana e antioxidante do extrato hidroetanólico de leonotis nepetaefolia (l) r. Br. |
title_full |
Avaliação da citotoxicidade e da atividade antimicrobiana e antioxidante do extrato hidroetanólico de leonotis nepetaefolia (l) r. Br. |
title_fullStr |
Avaliação da citotoxicidade e da atividade antimicrobiana e antioxidante do extrato hidroetanólico de leonotis nepetaefolia (l) r. Br. |
title_full_unstemmed |
Avaliação da citotoxicidade e da atividade antimicrobiana e antioxidante do extrato hidroetanólico de leonotis nepetaefolia (l) r. Br. |
title_sort |
Avaliação da citotoxicidade e da atividade antimicrobiana e antioxidante do extrato hidroetanólico de leonotis nepetaefolia (l) r. Br. |
author |
Oliveira, Darley Maria |
author_facet |
Oliveira, Darley Maria |
author_role |
author |
dc.contributor.none.fl_str_mv |
Martins, Domingos Tabajara de Oliveira http://lattes.cnpq.br/3794477872946546 Martins, Domingos Tabajara de Oliveira 109.726.923-04 http://lattes.cnpq.br/3794477872946546 Slhessarenko, Renata Dezengrini 006.990.439-13 http://lattes.cnpq.br/3790088995387532 109.726.923-04 Cruz, Alexandre Bella 480.657.909-25 http://lattes.cnpq.br/1596661935560930 Lopes, Lousa 879.675.298-04 http://lattes.cnpq.br/8843725522665416 |
dc.contributor.author.fl_str_mv |
Oliveira, Darley Maria |
dc.subject.por.fl_str_mv |
Leonotis nepetefolia Antimicrobiano Citotoxicidade Antioxidante CNPQ::CIENCIAS DA SAUDE::MEDICINA Leonotis nepetefolia Antimicrobial Cytotoxicity Antioxidant |
topic |
Leonotis nepetefolia Antimicrobiano Citotoxicidade Antioxidante CNPQ::CIENCIAS DA SAUDE::MEDICINA Leonotis nepetefolia Antimicrobial Cytotoxicity Antioxidant |
description |
Leonotis nepetifolia (L.) R. Br (Lamiaceae), popularly known as “cordão-de- frade”, is a sub-spontaneous shrub. The tea from the whole plant is used in the treatment of infections, inflammations, wounds and stomach ulcers. The present study evaluates the potential antimicrobial, antioxidant and cytotoxicity of the hydroethanolic extract of L. nepetifolia (HELn). The whole plant was washed, dried, powdered and the resulting powder was macerated in hydroethanolic solution (1:5, w/v), filtered, concentrated in rotaryevaporator and the residual solvent was eliminated in hotair oven, thus obtaining HELn. Chinese hamster ovary cells (CHO-K1) were used for cytotoxicity studies by Alamar blue method. To evaluate the antimicrobial activity against a panel of Gram-positive and Gram-negative bacteria and yeasts and filamentos fungi, disk diffusion test and broth microdilution were used. Drugs with inhibition zones of ≥ 10 mm and minimal inhibitory concentration (MIC) <100 µg/mL were considered very active. To characterize the nature of the antibacterial activity, antibiosis mechanism study was conducted, evaluating the growth of sensitive bacteria to concentrations of HELn ≥ MIC within 24 h. To assess the antibacterial mode of action of HELn on the permeability of the outer membrane, synergism between the extract (0.5 MIC) and hydrophobic antibiotics- erythromycin and rifampicin (0.68 to 20 µg/mL), was determined in Shigella flexneri. However, the mode of action of of HELn on the cytoplasmic membrane was evaluated by measuring the potassium efflux (2 MIC) and nucleotides leakages (1 MIC), using electrolytes analyzer and spectrophotometer respectively, in strains of E. faecalis and S. flexneri. The antioxidant activity was determined in vitro by DPPH, NO and H2O2 radical scavenging assays. The IC50 of HELn in the cytotoxicity assay was greater than 200 µg/mL. HELn demostrated a good spectrum of antibacterial activity in broth microdilution assay against S. flexneri, S. aureus and E. faecalis (MIC= 6.25 µg/mL), H. pylori (MIC= 25 µg/mL) and S. pyogenes (MIC= 50 µg/mL). With chloramphenicol, the MIC varied between 3.12 µg/mL (S. flexneri ) and 100 µg/mL (P. aeruginosa). In the disk diffusion assay, HELn was inactive, while chloramphenicol produced inhibition halos of 30 mm against all strains tested. HELn presented a bacteriostatic action against the sensitive bacterial strains. In the presence of HELn, S. flexneri became more sensitive to the probe antibiotics at MIC of erythromycin (1 µg/mL) and below the MIC of rifampicin (5 µg/mL). HELn caused an intense damage to the cytoplasmic membrane, reaching a maximum K+ efflux of 11.7 ± 0.6 ppm and 12.87 ± 0.3 ppm, and of the nucleotide 22% (6th hour) and 46% (12th time ) in E. faecalis and S. flexneri, respectively. EHLn had no effect on fungal growth, unlike amphotericin B, which was active in both models of disk diffusion and broth microdilution, attaining maxima activity against yeasts (35 mm) and filamentous fungi (3.12 µg/mL). HELn demonstrated weak antioxidant activity with IC50 (µg/mL) of 128.94 ± 8.7, 190.93 ± 0.01 and > 250 for DPPH, NO and H2O2 radical scavenging assays respectively. The results indicate that HELn has low cytotoxicity and antioxidant activity and potent antibacterial activity, especially against Gram-positive bacteria,. It is bacteriostatic in nature, possibly acting at the level of the cell wall and cytoplasmic membrane thus supporting its popular use in infectious processes. In addition, the presence of phenols, flavonoides, carotenoids, fatty acids and esteroids, described in the literature to posses antioxidant and antimicrobial, were detected in the composition of HELn |
publishDate |
2013 |
dc.date.none.fl_str_mv |
2013-06-05 2013-05-23 2019-11-21T13:29:55Z 2019-11-21T13:29:55Z |
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 |
OLIVEIRA, Darley Maria. Avaliação da citotoxicidade e da atividade antimicrobiana e antioxidante do extrato hidroetanólico de leonotis nepetaefolia (l) r. Br. 2013. 96 f. Dissertação (Mestrado em Ciências da Saúde) - Universidade Federal de Mato Grosso, Faculdade de Medicina, Cuiabá, 2013. http://ri.ufmt.br/handle/1/1590 |
identifier_str_mv |
OLIVEIRA, Darley Maria. Avaliação da citotoxicidade e da atividade antimicrobiana e antioxidante do extrato hidroetanólico de leonotis nepetaefolia (l) r. Br. 2013. 96 f. Dissertação (Mestrado em Ciências da Saúde) - Universidade Federal de Mato Grosso, Faculdade de Medicina, Cuiabá, 2013. |
url |
http://ri.ufmt.br/handle/1/1590 |
dc.language.iso.fl_str_mv |
por |
language |
por |
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info:eu-repo/semantics/openAccess |
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openAccess |
dc.publisher.none.fl_str_mv |
Universidade Federal de Mato Grosso Brasil Faculdade de Medicina (FM) UFMT CUC - Cuiabá Programa de Pós-Graduação em Ciências da Saúde |
publisher.none.fl_str_mv |
Universidade Federal de Mato Grosso Brasil Faculdade de Medicina (FM) UFMT CUC - Cuiabá Programa de Pós-Graduação em Ciências da Saúde |
dc.source.none.fl_str_mv |
reponame:Repositório Institucional da UFMT instname:Universidade Federal de Mato Grosso (UFMT) instacron:UFMT |
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Universidade Federal de Mato Grosso (UFMT) |
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UFMT |
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UFMT |
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Repositório Institucional da UFMT |
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Repositório Institucional da UFMT |
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Repositório Institucional da UFMT - Universidade Federal de Mato Grosso (UFMT) |
repository.mail.fl_str_mv |
jordanbiblio@gmail.com |
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1800846262754344960 |