Efeito dos carotenoides do urucum sobre a fragilidade eritrocitária in vitro e ex vivo
Autor(a) principal: | |
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Data de Publicação: | 2019 |
Tipo de documento: | Dissertação |
Idioma: | por |
Título da fonte: | Manancial - Repositório Digital da UFSM |
Texto Completo: | http://repositorio.ufsm.br/handle/1/21580 |
Resumo: | Erythrocytes are the cells with the main function of transporting respiratory gases to tissues. These cells exhibit high susceptibility to hemolysis in various pathologies due to oxidation of their cellular components. Under physiological conditions, the production of reactive oxygen species (ROS) is controlled by the endogenous antioxidant system, continuously maintaining balance. However, an overproduction of ROS results in an imbalance between the pro and antioxidant system, called oxidative stress. The carotenoids bixin (BIX) and norbixin (NBIX) are mainly found in annatto seeds (Bixa orellana L.). These compounds have excellent antioxidant capacity and anti-inflammatory action. The aim of this study was to evaluate the antioxidant activity and the ability of BIX and NBIX to protect erythrocytes from in vitro and ex vivo induced hemolysis. Healthy adults of both sexes participated in the study. Erythrocytes from 4 volunteers were isolated for the in vitro experiment and co-incubated with different concentrations of bixin or norbixin and three different inducers of cell fragility. In the ex vivo study, 8 volunteers received a daily capsule containing BIX or NBIX (0.5 mg/kg/day) or placebo for 7 days, in a crossover design. The randomized, double-blind, placebo-controlled clinical trial was performed using a 21-day washout period between treatments, so that all volunteers received all treatments at different times. After the last day of each treatment, blood was collected. Erythrocytes were isolated and incubated with three different cell damage inducers (2,2'-azobis (2-amidinopropane) dihydrochloride (AAPH), glucose and NaNO2), which act by different mechanisms. In both the ex vivo and in vitro studies, after incubation with the inducers, the erythrocytes were submitted to osmotic fragility tests, where the erythrocytes were exposed to different NaCl concentrations (0.2 to 0.9%), with erythrocyte resistance being evaluated later. Moreover, in the ex vivo experiment, the activity of catalase, superoxide dismutase and glutathione peroxidase, as well as the levels of lipid oxidation malondialdehyde (MDA) were evaluated. In vitro, BIX and NBIX not only reduced AAPH, glucose or NaNO2-induced erythrocyte membrane fragility, but also improved basal osmotic resistance in the micromolar range. BIX and NBIX supplementation significantly increased erythrocyte membrane resistance, and BIX supplementation was the most effective. In addition, BIX and NBIX protected erythrocytes from lipid peroxidation and improved the cellular redox environment. Our study demonstrates the antihemolytic potential of annatto carotenoids by preventing and/or reducing membrane fragility of human erythrocytes. Additionally, it contributes to broaden understanding of the beneficial properties of human consumption of carotenoids, especially BIX and NBIX. |
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Efeito dos carotenoides do urucum sobre a fragilidade eritrocitária in vitro e ex vivoEffect of urucum carotenoids on eritrocyte fragility in vitro and ex vivoPotencial anti-hemolíticoBixinaNorbixinaEnsaio clínicoAnemiaHemáciasAntihemolytic potentialBixinNorbixinClinical trialRed blood cellsCNPQ::CIENCIAS BIOLOGICAS::FARMACOLOGIAErythrocytes are the cells with the main function of transporting respiratory gases to tissues. These cells exhibit high susceptibility to hemolysis in various pathologies due to oxidation of their cellular components. Under physiological conditions, the production of reactive oxygen species (ROS) is controlled by the endogenous antioxidant system, continuously maintaining balance. However, an overproduction of ROS results in an imbalance between the pro and antioxidant system, called oxidative stress. The carotenoids bixin (BIX) and norbixin (NBIX) are mainly found in annatto seeds (Bixa orellana L.). These compounds have excellent antioxidant capacity and anti-inflammatory action. The aim of this study was to evaluate the antioxidant activity and the ability of BIX and NBIX to protect erythrocytes from in vitro and ex vivo induced hemolysis. Healthy adults of both sexes participated in the study. Erythrocytes from 4 volunteers were isolated for the in vitro experiment and co-incubated with different concentrations of bixin or norbixin and three different inducers of cell fragility. In the ex vivo study, 8 volunteers received a daily capsule containing BIX or NBIX (0.5 mg/kg/day) or placebo for 7 days, in a crossover design. The randomized, double-blind, placebo-controlled clinical trial was performed using a 21-day washout period between treatments, so that all volunteers received all treatments at different times. After the last day of each treatment, blood was collected. Erythrocytes were isolated and incubated with three different cell damage inducers (2,2'-azobis (2-amidinopropane) dihydrochloride (AAPH), glucose and NaNO2), which act by different mechanisms. In both the ex vivo and in vitro studies, after incubation with the inducers, the erythrocytes were submitted to osmotic fragility tests, where the erythrocytes were exposed to different NaCl concentrations (0.2 to 0.9%), with erythrocyte resistance being evaluated later. Moreover, in the ex vivo experiment, the activity of catalase, superoxide dismutase and glutathione peroxidase, as well as the levels of lipid oxidation malondialdehyde (MDA) were evaluated. In vitro, BIX and NBIX not only reduced AAPH, glucose or NaNO2-induced erythrocyte membrane fragility, but also improved basal osmotic resistance in the micromolar range. BIX and NBIX supplementation significantly increased erythrocyte membrane resistance, and BIX supplementation was the most effective. In addition, BIX and NBIX protected erythrocytes from lipid peroxidation and improved the cellular redox environment. Our study demonstrates the antihemolytic potential of annatto carotenoids by preventing and/or reducing membrane fragility of human erythrocytes. Additionally, it contributes to broaden understanding of the beneficial properties of human consumption of carotenoids, especially BIX and NBIX.Os eritrócitos são as células com a principal função de transportar gases respiratórios para os tecidos. Essas células exibem grande suscetibilidade à hemólise em diversas patologias, devido à oxidação de seus componentes celulares. Em situações fisiológicas normais, a produção de espécies reativas de oxigênio (EROs) é controlada pelo sistema antioxidante endógeno, mantendo continuamente o equilíbrio. No entanto, uma superprodução de ROS resulta em desequilíbrio entre o sistema pró e antioxidante, o chamado estresse oxidativo. Os carotenoides bixina (BIX) e norbixina (NBIX) são encontrados principalmente na semente do urucum (Bixa orellana L.). Estes compostos possuem excelente capacidade antioxidante e ação antiinflamatória. O objetivo deste estudo foi avaliar a atividade antioxidante e a capacidade da BIX e da NBIX em proteger os eritrócitos da hemólise induzida in vitro e ex vivo. Adultos saudáveis de ambos os sexos participaram do estudo, sendo utilizado para o experimento in vitro eritrócitos de 4 voluntários, que foram isolados e co-incubados com diferentes concentrações de BIX ou NBIX e três diferentes indutores de fragilidade celular. No estudo ex vivo com delineamento cruzado com retorno, 8 voluntários receberam diariamente uma cápsula contendo BIX ou NBIX (0,05 mg/kg/dia) ou placebo durante sete dias. O ensaio clínico foi randomizado, duplo-cego, controlado por placebo e empregou-se washout de 21 dias entre os tratamentos, de modo que todos os voluntários receberam todos os tratamentos em momentos diferentes. Após o último dia de cada tratamento, o sangue foi coletado. Os eritrócitos foram isolados e incubados com três diferentes indutores de dano celular (dicloridrato de 2,2'-azobis (2- amidinopropano) (AAPH), glicose e NaNO2), que atuam por mecanismos distintos. Tanto no estudo ex vivo quanto no in vitro, após a incubação com os indutores, os eritrócitos foram submetidos aos testes de fragilidade osmótica, onde os eritrócitos foram expostos a diferentes concentrações de NaCl (0,2% a 0,9%), seguido da avaliação do grau de hemólise como indicador da resistência dos eritrócitos. Além disso, no experimento ex vivo foi avaliada a atividade das enzimas catalase, superóxido dismutase e glutationa peroxidase, bem como os níveis de malondialdeído (MDA), produto da oxidação lipídica. In vitro, BIX e NBIX não apenas reduziram a fragilidade da membrana eritrocitária induzida por AAPH, glicose ou NaNO2, mas também melhoraram a resistência osmótica basal em concentrações micromolares. A suplementação com BIX ou NBIX aumentou significativamente a resistência da membrana eritrocitária, e a suplementação com BIX foi a mais efetiva. Além disso, BIX e NBIX protegeram os eritrócitos da peroxidação lipídica e melhoraram o ambiente redox celular. Nosso estudo demostra o potencial anti-hemolítico dos carotenoides do urucum, ao impedir e/ou reduzir a fragilidade da membrana de eritrócitos humanos. Adicionalmente, contribui para ampliar o entendimento acerca das propriedades benéficas do consumo humano de carotenoides, especialmente a BIX e a NBIX.Universidade Federal de Santa MariaBrasilFarmacologiaUFSMPrograma de Pós-Graduação em FarmacologiaCentro de Ciências da SaúdeEmanuelli, Tatianahttp://lattes.cnpq.br/2165391096880394Somacal, SabrinaDenardin, Cristiane CasagrandeChitolina, Maria RosaBeni, Anissa Algara2021-07-23T11:38:31Z2021-07-23T11:38:31Z2019-08-30info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisapplication/pdfhttp://repositorio.ufsm.br/handle/1/21580porAttribution-NonCommercial-NoDerivatives 4.0 Internationalhttp://creativecommons.org/licenses/by-nc-nd/4.0/info:eu-repo/semantics/openAccessreponame:Manancial - Repositório Digital da UFSMinstname:Universidade Federal de Santa Maria (UFSM)instacron:UFSM2021-07-24T06:01:52Zoai:repositorio.ufsm.br:1/21580Biblioteca Digital de Teses e Dissertaçõeshttps://repositorio.ufsm.br/ONGhttps://repositorio.ufsm.br/oai/requestatendimento.sib@ufsm.br||tedebc@gmail.comopendoar:2021-07-24T06:01:52Manancial - Repositório Digital da UFSM - Universidade Federal de Santa Maria (UFSM)false |
dc.title.none.fl_str_mv |
Efeito dos carotenoides do urucum sobre a fragilidade eritrocitária in vitro e ex vivo Effect of urucum carotenoids on eritrocyte fragility in vitro and ex vivo |
title |
Efeito dos carotenoides do urucum sobre a fragilidade eritrocitária in vitro e ex vivo |
spellingShingle |
Efeito dos carotenoides do urucum sobre a fragilidade eritrocitária in vitro e ex vivo Beni, Anissa Algara Potencial anti-hemolítico Bixina Norbixina Ensaio clínico Anemia Hemácias Antihemolytic potential Bixin Norbixin Clinical trial Red blood cells CNPQ::CIENCIAS BIOLOGICAS::FARMACOLOGIA |
title_short |
Efeito dos carotenoides do urucum sobre a fragilidade eritrocitária in vitro e ex vivo |
title_full |
Efeito dos carotenoides do urucum sobre a fragilidade eritrocitária in vitro e ex vivo |
title_fullStr |
Efeito dos carotenoides do urucum sobre a fragilidade eritrocitária in vitro e ex vivo |
title_full_unstemmed |
Efeito dos carotenoides do urucum sobre a fragilidade eritrocitária in vitro e ex vivo |
title_sort |
Efeito dos carotenoides do urucum sobre a fragilidade eritrocitária in vitro e ex vivo |
author |
Beni, Anissa Algara |
author_facet |
Beni, Anissa Algara |
author_role |
author |
dc.contributor.none.fl_str_mv |
Emanuelli, Tatiana http://lattes.cnpq.br/2165391096880394 Somacal, Sabrina Denardin, Cristiane Casagrande Chitolina, Maria Rosa |
dc.contributor.author.fl_str_mv |
Beni, Anissa Algara |
dc.subject.por.fl_str_mv |
Potencial anti-hemolítico Bixina Norbixina Ensaio clínico Anemia Hemácias Antihemolytic potential Bixin Norbixin Clinical trial Red blood cells CNPQ::CIENCIAS BIOLOGICAS::FARMACOLOGIA |
topic |
Potencial anti-hemolítico Bixina Norbixina Ensaio clínico Anemia Hemácias Antihemolytic potential Bixin Norbixin Clinical trial Red blood cells CNPQ::CIENCIAS BIOLOGICAS::FARMACOLOGIA |
description |
Erythrocytes are the cells with the main function of transporting respiratory gases to tissues. These cells exhibit high susceptibility to hemolysis in various pathologies due to oxidation of their cellular components. Under physiological conditions, the production of reactive oxygen species (ROS) is controlled by the endogenous antioxidant system, continuously maintaining balance. However, an overproduction of ROS results in an imbalance between the pro and antioxidant system, called oxidative stress. The carotenoids bixin (BIX) and norbixin (NBIX) are mainly found in annatto seeds (Bixa orellana L.). These compounds have excellent antioxidant capacity and anti-inflammatory action. The aim of this study was to evaluate the antioxidant activity and the ability of BIX and NBIX to protect erythrocytes from in vitro and ex vivo induced hemolysis. Healthy adults of both sexes participated in the study. Erythrocytes from 4 volunteers were isolated for the in vitro experiment and co-incubated with different concentrations of bixin or norbixin and three different inducers of cell fragility. In the ex vivo study, 8 volunteers received a daily capsule containing BIX or NBIX (0.5 mg/kg/day) or placebo for 7 days, in a crossover design. The randomized, double-blind, placebo-controlled clinical trial was performed using a 21-day washout period between treatments, so that all volunteers received all treatments at different times. After the last day of each treatment, blood was collected. Erythrocytes were isolated and incubated with three different cell damage inducers (2,2'-azobis (2-amidinopropane) dihydrochloride (AAPH), glucose and NaNO2), which act by different mechanisms. In both the ex vivo and in vitro studies, after incubation with the inducers, the erythrocytes were submitted to osmotic fragility tests, where the erythrocytes were exposed to different NaCl concentrations (0.2 to 0.9%), with erythrocyte resistance being evaluated later. Moreover, in the ex vivo experiment, the activity of catalase, superoxide dismutase and glutathione peroxidase, as well as the levels of lipid oxidation malondialdehyde (MDA) were evaluated. In vitro, BIX and NBIX not only reduced AAPH, glucose or NaNO2-induced erythrocyte membrane fragility, but also improved basal osmotic resistance in the micromolar range. BIX and NBIX supplementation significantly increased erythrocyte membrane resistance, and BIX supplementation was the most effective. In addition, BIX and NBIX protected erythrocytes from lipid peroxidation and improved the cellular redox environment. Our study demonstrates the antihemolytic potential of annatto carotenoids by preventing and/or reducing membrane fragility of human erythrocytes. Additionally, it contributes to broaden understanding of the beneficial properties of human consumption of carotenoids, especially BIX and NBIX. |
publishDate |
2019 |
dc.date.none.fl_str_mv |
2019-08-30 2021-07-23T11:38:31Z 2021-07-23T11:38:31Z |
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 |
http://repositorio.ufsm.br/handle/1/21580 |
url |
http://repositorio.ufsm.br/handle/1/21580 |
dc.language.iso.fl_str_mv |
por |
language |
por |
dc.rights.driver.fl_str_mv |
Attribution-NonCommercial-NoDerivatives 4.0 International http://creativecommons.org/licenses/by-nc-nd/4.0/ info:eu-repo/semantics/openAccess |
rights_invalid_str_mv |
Attribution-NonCommercial-NoDerivatives 4.0 International http://creativecommons.org/licenses/by-nc-nd/4.0/ |
eu_rights_str_mv |
openAccess |
dc.format.none.fl_str_mv |
application/pdf |
dc.publisher.none.fl_str_mv |
Universidade Federal de Santa Maria Brasil Farmacologia UFSM Programa de Pós-Graduação em Farmacologia Centro de Ciências da Saúde |
publisher.none.fl_str_mv |
Universidade Federal de Santa Maria Brasil Farmacologia UFSM Programa de Pós-Graduação em Farmacologia Centro de Ciências da Saúde |
dc.source.none.fl_str_mv |
reponame:Manancial - Repositório Digital da UFSM instname:Universidade Federal de Santa Maria (UFSM) instacron:UFSM |
instname_str |
Universidade Federal de Santa Maria (UFSM) |
instacron_str |
UFSM |
institution |
UFSM |
reponame_str |
Manancial - Repositório Digital da UFSM |
collection |
Manancial - Repositório Digital da UFSM |
repository.name.fl_str_mv |
Manancial - Repositório Digital da UFSM - Universidade Federal de Santa Maria (UFSM) |
repository.mail.fl_str_mv |
atendimento.sib@ufsm.br||tedebc@gmail.com |
_version_ |
1805922110134026240 |