Avaliação in vitro da atividade anti-Helicobacter pylori e potencial antioxidante de extratos e frações de Baccharis trimera (Less.) DC
Autor(a) principal: | |
---|---|
Data de Publicação: | 2016 |
Tipo de documento: | Dissertação |
Idioma: | por |
Título da fonte: | Repositório Institucional da Universidade Federal do Espírito Santo (riUfes) |
Texto Completo: | http://repositorio.ufes.br/handle/10/5308 |
Resumo: | One of the major risk factors for the development of ulcers and gastric cancer is the infection by Helicobacter pylori, in which there is a considerable oxidative stress. In Brazil, the plant Baccharis trimera (Less) DC, known as "Carqueja", is commonly used to treat gastrointestinal and liver disorders. Tannins, flavonoids, saponins, diterpene lactones, sesquiterpenes, phenolic compounds and volatile oils were identified in this specie. Thus, the goal of this study was to evaluate the antiHelicobacter pylori activity of the aqueous (AqE), ethanolic (EE), and hydroalcoholic (HE) extracts of B. trimera, as well as the aqueous (AqF), hexane (HxF), and acetonitrile / chloroform (ACF) fractions obtained from the HE. In addition, it aims to analyze the phytochemical composition and its effects on free radicals and biological oxidants. For all extracts and fractions, it was performed a phytochemical screening and the determination of the content of polyphenol, flavonoid and total tannins. The anti- Helicobacter pylori activity was evaluated by microdilution broth assay and urease inhibitory capacity. Bacterial morphology, after sample exposure, was evaluated by scanning electron microscope (SEM) using the extracts and fractions that reached the minimum inhibitory concentration (CIM90). Screening of antioxidant activity for all samples was determined by DPPH and ABTS radical scavenging activities. Assays using the oxidizing biological agents, hydrogen peroxide (H2O2), superoxide anion (O2 •- ), hypochlorous acid (HOCl), hydroxyl radical (HO• ), and nitric oxide (NO• ) were performed with HE and its fractions due the best results in the antiH. pylori and initial screening antioxidant assays, except for the HxF because of its poor performance in the antioxidant screening. The highest concentration of polyphenols was observed in the HE, flavonoids in the ACF, and tannins in the AqF. Anti-H pylori assay presented a CIM90 of 512 μg/mL for HE, and 1024 μg/mL for ACF, the latter being bactericidal. SEM showed morphological changes such as stretching and cell lysis in the samples tested, and may suggest some modifications in the wall, such as alterations in the peptidoglycan synthesis. In the enzyme urease inhibition assay, the most significant result was obtained by the ACF, which inhibited 36.24% in the highest concentration tested. The best result among the extracts in the initial antioxidant screening was achieved by the HE, which obtained the following EC50: DPPH, 17.40 ± 0.52 and ABTS, 9.99 ± 1.21. The best EC50 for the fractions were: DPPH, AqF, 27.41 ± 1.65, and ABTS, AqF, 10.80 ± 1.90. In the O2 •- scavenging activity, AqF showed EC50 of 5.85 ± 0.86 μg/mL. In the inhibition assay of HOCl, all samples tested were able to inhibit greater than 50%, except AqF. In the inhibition assay of HO• , NO• e H2O2, the results were better for ACF and its EC50 was 2.90 ± 0.48, 132.13 ± 7.38 e 66.70 ± 2.30 μg/mL, respectively. The analyzes indicate that B. trimera, in particular the HE and its fractions, AqF and ACF, may exhibit promising compounds for the prevention and treatment of diseases caused by H. pylori. These results were based on the relationship between the presence of phenolic compounds and the inhibition of oxidants, as well as changes in the bacterial membrane. |
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Gonçalves, Rita de Cássia RibeiroNunes, Otalibio CastiglioniVellosa, José Carlos RebuglioBorges, Warley de Souza2016-08-29T15:38:48Z2016-07-112016-08-29T15:38:48Z2016-03-22One of the major risk factors for the development of ulcers and gastric cancer is the infection by Helicobacter pylori, in which there is a considerable oxidative stress. In Brazil, the plant Baccharis trimera (Less) DC, known as "Carqueja", is commonly used to treat gastrointestinal and liver disorders. Tannins, flavonoids, saponins, diterpene lactones, sesquiterpenes, phenolic compounds and volatile oils were identified in this specie. Thus, the goal of this study was to evaluate the antiHelicobacter pylori activity of the aqueous (AqE), ethanolic (EE), and hydroalcoholic (HE) extracts of B. trimera, as well as the aqueous (AqF), hexane (HxF), and acetonitrile / chloroform (ACF) fractions obtained from the HE. In addition, it aims to analyze the phytochemical composition and its effects on free radicals and biological oxidants. For all extracts and fractions, it was performed a phytochemical screening and the determination of the content of polyphenol, flavonoid and total tannins. The anti- Helicobacter pylori activity was evaluated by microdilution broth assay and urease inhibitory capacity. Bacterial morphology, after sample exposure, was evaluated by scanning electron microscope (SEM) using the extracts and fractions that reached the minimum inhibitory concentration (CIM90). Screening of antioxidant activity for all samples was determined by DPPH and ABTS radical scavenging activities. Assays using the oxidizing biological agents, hydrogen peroxide (H2O2), superoxide anion (O2 •- ), hypochlorous acid (HOCl), hydroxyl radical (HO• ), and nitric oxide (NO• ) were performed with HE and its fractions due the best results in the antiH. pylori and initial screening antioxidant assays, except for the HxF because of its poor performance in the antioxidant screening. The highest concentration of polyphenols was observed in the HE, flavonoids in the ACF, and tannins in the AqF. Anti-H pylori assay presented a CIM90 of 512 μg/mL for HE, and 1024 μg/mL for ACF, the latter being bactericidal. SEM showed morphological changes such as stretching and cell lysis in the samples tested, and may suggest some modifications in the wall, such as alterations in the peptidoglycan synthesis. In the enzyme urease inhibition assay, the most significant result was obtained by the ACF, which inhibited 36.24% in the highest concentration tested. The best result among the extracts in the initial antioxidant screening was achieved by the HE, which obtained the following EC50: DPPH, 17.40 ± 0.52 and ABTS, 9.99 ± 1.21. The best EC50 for the fractions were: DPPH, AqF, 27.41 ± 1.65, and ABTS, AqF, 10.80 ± 1.90. In the O2 •- scavenging activity, AqF showed EC50 of 5.85 ± 0.86 μg/mL. In the inhibition assay of HOCl, all samples tested were able to inhibit greater than 50%, except AqF. In the inhibition assay of HO• , NO• e H2O2, the results were better for ACF and its EC50 was 2.90 ± 0.48, 132.13 ± 7.38 e 66.70 ± 2.30 μg/mL, respectively. The analyzes indicate that B. trimera, in particular the HE and its fractions, AqF and ACF, may exhibit promising compounds for the prevention and treatment of diseases caused by H. pylori. These results were based on the relationship between the presence of phenolic compounds and the inhibition of oxidants, as well as changes in the bacterial membrane.Um dos principais fatores de risco para o desenvolvimento de úlceras e câncer gástrico é a infecção pela bactéria Helicobacter pylori, que é acompanhada por um considerável estresse oxidativo. No Brasil, a planta Baccharis trimera Less. (DC), conhecida como carqueja, é popularmente usada para tratar distúrbios gastrointestinais e do fígado. Nessa espécie já foram identificados taninos, flavonoides, saponinas, lactonas diterpênicas, sesquiterpenos, compostos fenólicos e óleos voláteis. Dessa forma, o objetivo do presente estudo foi avaliar a atividade anti-Helicobacter pylori do extrato aquoso, (EAq), etanólico (EE), hidroalcoólico (EH) de B. trimera e das frações, aquosa (FAq), hexânica (FHx), e acetonitrila/clorofórmio (FAC) obtidas a partir do EH, além de analisar a composição fitoquímica, e seus efeitos sobre radicais livres e oxidantes biológicos. Para todos os extratos e frações foi realizada a triagem fitoquímica e a determinação do teor de polifenóis, flavonoides e taninos totais. A atividade anti-Helicobacter pylori foi avaliada por meio do ensaio de microdiluição em caldo e pela capacidade de inibição da enzima urease. A morfologia bacteriana, após exposição às amostras, foi analisada por microscópio eletrônico de varredura (MEV) com o extrato e a fração que atingiram a concentração inibitória mínima (CIM90). A atividade antioxidante foi avaliada em ensaios de inibição de radicais sintéticos DPPH e ABTS como triagem inicial para todas as amostras. Os ensaios com os oxidantes biológicos, peróxido de hidrogênio (H2O2), ânion superóxido (O2 -), ácido hipocloroso (HOCl), radical hidroxil (HO ) e óxido nítrico (NO ), foram realizados com o EH, devido aos melhores resultados obtidos no ensaio anti-H.pylori e na triagem antioxidante inicial, além de suas frações, exceto a FHx, devido ao baixo desempenho na triagem antioxidante. A maior concentração de polifenóis foi encontrado no EH, flavonoides na FAC e taninos em FAq. No ensaio anti-H. pylori, a CIM90 foi de 512 μg/mL para EH e 1024 μg/mL para FAC a qual foi bactericida. A MEV mostrou alterações morfológicas como alongamento e lise celular nas amostras testadas, podendo sugerir alterações de parede, provavelmente através do comprometimento da síntese de peptideoglicano. No ensaio de inibição da enzima urease o resultado mais expressivo foi o exibido pela FAC, que inibiu 36,24% na maior concentração testada.O melhor resultado dentre os extratos na triagem antioxidante inicial foi do EH, que obteve os seguintes CE50: DPPH, 17,40 ± 0,52 e ABTS, 9,99 ± 1,21. Para as frações os melhores CE50 foram: DPPH, FAq, 27,41 ± 1,65 e ABTS, FAq, 10,80 ± 1,90.No ensaio de captura do O2 -, a FAq apresentou CE50 de 5,85 ± 0,86μg/mL. No ensaio de inibição do HOCl, todas as amostras testadas conseguiram inibição maior que 50%, exceto a FAq. No ensaio de inibição do radical HO , NO e H2O2, os resultados foram melhores para FAC e seu CE50 foi de 2,90 ± 0,48, 132,13 ± 7,38 e 66,70 ± 2,30 μg/mL, respectivamente. As análises indicam que B. trimera, especialmente o EH, e suas frações, FAq e FAC, podem apresentar compostos promissores para o prevenção e tratamento de doenças causadas por H. pylori. Estes resultados foram embasados pela relação entre a presença de compostos fenólicos e a inibição de oxidantes, além da observação de alterações na membrana bacteriana.Texthttp://repositorio.ufes.br/handle/10/5308porUniversidade Federal do Espírito SantoMestrado em Ciências FarmacêuticasPrograma de Pós-Graduação em Ciências FarmacêuticasUFESBRCentro de Ciências da SaúdeOxidative stressHelicobacter pyloriBaccharis trimeraEstresse oxidativoCompostos fenólicosAção antioxidanteStress oxidativoFenóisAntioxidantesFarmácia615Avaliação in vitro da atividade anti-Helicobacter pylori e potencial antioxidante de extratos e frações de Baccharis trimera (Less.) DCinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisinfo:eu-repo/semantics/openAccessreponame:Repositório Institucional da Universidade Federal do Espírito Santo (riUfes)instname:Universidade Federal do Espírito Santo (UFES)instacron:UFESORIGINALtese_8529_DISSERTAÇÃO MESTRADO 101 DEFINITIVA.pdfapplication/pdf1899694http://repositorio.ufes.br/bitstreams/0e1f13a7-bfa5-4699-b399-d5bdd3208017/download2efb55c7dbd8ad135bab9068c1763cebMD5110/53082024-07-16 17:09:38.686oai:repositorio.ufes.br:10/5308http://repositorio.ufes.brRepositório InstitucionalPUBhttp://repositorio.ufes.br/oai/requestopendoar:21082024-10-15T17:57:50.791104Repositório Institucional da Universidade Federal do Espírito Santo (riUfes) - Universidade Federal do Espírito Santo (UFES)false |
dc.title.none.fl_str_mv |
Avaliação in vitro da atividade anti-Helicobacter pylori e potencial antioxidante de extratos e frações de Baccharis trimera (Less.) DC |
title |
Avaliação in vitro da atividade anti-Helicobacter pylori e potencial antioxidante de extratos e frações de Baccharis trimera (Less.) DC |
spellingShingle |
Avaliação in vitro da atividade anti-Helicobacter pylori e potencial antioxidante de extratos e frações de Baccharis trimera (Less.) DC Nunes, Otalibio Castiglioni Oxidative stress Helicobacter pylori Baccharis trimera Estresse oxidativo Compostos fenólicos Ação antioxidante Farmácia Stress oxidativo Fenóis Antioxidantes 615 |
title_short |
Avaliação in vitro da atividade anti-Helicobacter pylori e potencial antioxidante de extratos e frações de Baccharis trimera (Less.) DC |
title_full |
Avaliação in vitro da atividade anti-Helicobacter pylori e potencial antioxidante de extratos e frações de Baccharis trimera (Less.) DC |
title_fullStr |
Avaliação in vitro da atividade anti-Helicobacter pylori e potencial antioxidante de extratos e frações de Baccharis trimera (Less.) DC |
title_full_unstemmed |
Avaliação in vitro da atividade anti-Helicobacter pylori e potencial antioxidante de extratos e frações de Baccharis trimera (Less.) DC |
title_sort |
Avaliação in vitro da atividade anti-Helicobacter pylori e potencial antioxidante de extratos e frações de Baccharis trimera (Less.) DC |
author |
Nunes, Otalibio Castiglioni |
author_facet |
Nunes, Otalibio Castiglioni |
author_role |
author |
dc.contributor.advisor1.fl_str_mv |
Gonçalves, Rita de Cássia Ribeiro |
dc.contributor.author.fl_str_mv |
Nunes, Otalibio Castiglioni |
dc.contributor.referee1.fl_str_mv |
Vellosa, José Carlos Rebuglio |
dc.contributor.referee2.fl_str_mv |
Borges, Warley de Souza |
contributor_str_mv |
Gonçalves, Rita de Cássia Ribeiro Vellosa, José Carlos Rebuglio Borges, Warley de Souza |
dc.subject.eng.fl_str_mv |
Oxidative stress |
topic |
Oxidative stress Helicobacter pylori Baccharis trimera Estresse oxidativo Compostos fenólicos Ação antioxidante Farmácia Stress oxidativo Fenóis Antioxidantes 615 |
dc.subject.por.fl_str_mv |
Helicobacter pylori Baccharis trimera Estresse oxidativo Compostos fenólicos Ação antioxidante |
dc.subject.cnpq.fl_str_mv |
Farmácia |
dc.subject.br-rjbn.none.fl_str_mv |
Stress oxidativo Fenóis Antioxidantes |
dc.subject.udc.none.fl_str_mv |
615 |
description |
One of the major risk factors for the development of ulcers and gastric cancer is the infection by Helicobacter pylori, in which there is a considerable oxidative stress. In Brazil, the plant Baccharis trimera (Less) DC, known as "Carqueja", is commonly used to treat gastrointestinal and liver disorders. Tannins, flavonoids, saponins, diterpene lactones, sesquiterpenes, phenolic compounds and volatile oils were identified in this specie. Thus, the goal of this study was to evaluate the antiHelicobacter pylori activity of the aqueous (AqE), ethanolic (EE), and hydroalcoholic (HE) extracts of B. trimera, as well as the aqueous (AqF), hexane (HxF), and acetonitrile / chloroform (ACF) fractions obtained from the HE. In addition, it aims to analyze the phytochemical composition and its effects on free radicals and biological oxidants. For all extracts and fractions, it was performed a phytochemical screening and the determination of the content of polyphenol, flavonoid and total tannins. The anti- Helicobacter pylori activity was evaluated by microdilution broth assay and urease inhibitory capacity. Bacterial morphology, after sample exposure, was evaluated by scanning electron microscope (SEM) using the extracts and fractions that reached the minimum inhibitory concentration (CIM90). Screening of antioxidant activity for all samples was determined by DPPH and ABTS radical scavenging activities. Assays using the oxidizing biological agents, hydrogen peroxide (H2O2), superoxide anion (O2 •- ), hypochlorous acid (HOCl), hydroxyl radical (HO• ), and nitric oxide (NO• ) were performed with HE and its fractions due the best results in the antiH. pylori and initial screening antioxidant assays, except for the HxF because of its poor performance in the antioxidant screening. The highest concentration of polyphenols was observed in the HE, flavonoids in the ACF, and tannins in the AqF. Anti-H pylori assay presented a CIM90 of 512 μg/mL for HE, and 1024 μg/mL for ACF, the latter being bactericidal. SEM showed morphological changes such as stretching and cell lysis in the samples tested, and may suggest some modifications in the wall, such as alterations in the peptidoglycan synthesis. In the enzyme urease inhibition assay, the most significant result was obtained by the ACF, which inhibited 36.24% in the highest concentration tested. The best result among the extracts in the initial antioxidant screening was achieved by the HE, which obtained the following EC50: DPPH, 17.40 ± 0.52 and ABTS, 9.99 ± 1.21. The best EC50 for the fractions were: DPPH, AqF, 27.41 ± 1.65, and ABTS, AqF, 10.80 ± 1.90. In the O2 •- scavenging activity, AqF showed EC50 of 5.85 ± 0.86 μg/mL. In the inhibition assay of HOCl, all samples tested were able to inhibit greater than 50%, except AqF. In the inhibition assay of HO• , NO• e H2O2, the results were better for ACF and its EC50 was 2.90 ± 0.48, 132.13 ± 7.38 e 66.70 ± 2.30 μg/mL, respectively. The analyzes indicate that B. trimera, in particular the HE and its fractions, AqF and ACF, may exhibit promising compounds for the prevention and treatment of diseases caused by H. pylori. These results were based on the relationship between the presence of phenolic compounds and the inhibition of oxidants, as well as changes in the bacterial membrane. |
publishDate |
2016 |
dc.date.accessioned.fl_str_mv |
2016-08-29T15:38:48Z |
dc.date.available.fl_str_mv |
2016-07-11 2016-08-29T15:38:48Z |
dc.date.issued.fl_str_mv |
2016-03-22 |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
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info:eu-repo/semantics/masterThesis |
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http://repositorio.ufes.br/handle/10/5308 |
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por |
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Text |
dc.publisher.none.fl_str_mv |
Universidade Federal do Espírito Santo Mestrado em Ciências Farmacêuticas |
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Programa de Pós-Graduação em Ciências Farmacêuticas |
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UFES |
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BR |
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Centro de Ciências da Saúde |
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Universidade Federal do Espírito Santo Mestrado em Ciências Farmacêuticas |
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Universidade Federal do Espírito Santo (UFES) |
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UFES |
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Repositório Institucional da Universidade Federal do Espírito Santo (riUfes) |
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