Efeito da terapia de fotobiomodulação na expressão de biomarcadores de estresse oxidativo em células HUVEC submetidas à hipóxia

Detalhes bibliográficos
Ano de defesa: 2023
Autor(a) principal: Nasuk, Guilherme Rabelo lattes
Orientador(a): Silva Junior, Jose Antonio lattes
Banca de defesa: Silva Junior, Jose Antonio lattes, Zamuner, Stella Regina lattes, Ferrari, Raquel Agnelli Mesquita lattes
Tipo de documento: Dissertação
Tipo de acesso: Acesso aberto
Idioma: por
Instituição de defesa: Universidade Nove de Julho
Programa de Pós-Graduação: Programa de Mestrado em Medicina
Departamento: Saúde
País: Brasil
Palavras-chave em Português:
Palavras-chave em Inglês:
Área do conhecimento CNPq:
Link de acesso: http://bibliotecatede.uninove.br/handle/tede/3345
Resumo: Long considered to be an inert cell layer, the endothelium is now recognized as an important regulator of major physiological functions. Endothelial cells respond to chemical stimuli and secrete factors that modulate various physiological processes, especially under hypoxia. This condition can trigger adaptive responses and have harmful effects if it persists for prolonged periods, causing transcriptional changes that result in changes in cellular homeostasis. Photobiomodulation therapy (PBM) can stimulate collagen production, promote DNA and protein synthesis, increase ATP content, modulate cell migration and proliferation, accelerate tissue repair, and induce cell differentiation. In this context, the evaluation of cellular adaptations to hypoxia can provide knowledge to improve treatments and reduce these losses. This in vitro study aimed to evaluate the effects of pre-hypoxic PBM on human umbilical vein endothelial cells (HUVECs), to prevent possible damage caused by hypoxia. The methodology included gene expression and protein activity through in vitro tests. The cells were characterized by the expression of CD31 APC and CD146 APC by flow cytometry. The cells, kept in culture, were released from a 75cm2 matrix bottle and seeded in 25 cm² bottles at a quantity of 1x106. Our results showed that hypoxia only promoted a strong reduction in cell viability and that PBM applied pre-hypoxia increased this parameter. The mRNA expressions of the antioxidant enzymes SOD1 and CAT were reduced by hypoxia, and were increased with the use of PBM. The activities of SOD and GPx were increased by PBM. The expression of Hsp1a/b (HSP70), a cell damage biomarker, was increased by hypoxia, and this effect was blocked by the action of PBM, demonstrating HSP70 levels similar to the control. Increased levels of TBARS and free radicals were found in the hypoxia group compared to the control, interestingly, in the group previously treated with pre-hypoxic PBM therapy we observed a decrease in these oxidative stress markers, bringing values similar to the control. Together, the data suggest that PBM has the potential as a cell protection therapy against oxidative damage under hypoxic conditions, evidenced by the expression of antioxidant enzymes and the reduction of levels of carbonylated proteins and free radicals.
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spelling Silva Junior, Jose Antoniohttp://lattes.cnpq.br/1990427833073161Silva Junior, Jose Antoniohttp://lattes.cnpq.br/1990427833073161Zamuner, Stella Reginahttp://lattes.cnpq.br/1935805744318404Ferrari, Raquel Agnelli Mesquitahttp://lattes.cnpq.br/7048141683554943http://lattes.cnpq.br/1816053013460118Nasuk, Guilherme Rabelo2024-04-19T19:29:57Z2023-12-19Nasuk, Guilherme Rabelo. Efeito da terapia de fotobiomodulação na expressão de biomarcadores de estresse oxidativo em células HUVEC submetidas à hipóxia. 2023. 73 f. Dissertação( Programa de Mestrado em Medicina) - Universidade Nove de Julho, São Paulo.http://bibliotecatede.uninove.br/handle/tede/3345Long considered to be an inert cell layer, the endothelium is now recognized as an important regulator of major physiological functions. Endothelial cells respond to chemical stimuli and secrete factors that modulate various physiological processes, especially under hypoxia. This condition can trigger adaptive responses and have harmful effects if it persists for prolonged periods, causing transcriptional changes that result in changes in cellular homeostasis. Photobiomodulation therapy (PBM) can stimulate collagen production, promote DNA and protein synthesis, increase ATP content, modulate cell migration and proliferation, accelerate tissue repair, and induce cell differentiation. In this context, the evaluation of cellular adaptations to hypoxia can provide knowledge to improve treatments and reduce these losses. This in vitro study aimed to evaluate the effects of pre-hypoxic PBM on human umbilical vein endothelial cells (HUVECs), to prevent possible damage caused by hypoxia. The methodology included gene expression and protein activity through in vitro tests. The cells were characterized by the expression of CD31 APC and CD146 APC by flow cytometry. The cells, kept in culture, were released from a 75cm2 matrix bottle and seeded in 25 cm² bottles at a quantity of 1x106. Our results showed that hypoxia only promoted a strong reduction in cell viability and that PBM applied pre-hypoxia increased this parameter. The mRNA expressions of the antioxidant enzymes SOD1 and CAT were reduced by hypoxia, and were increased with the use of PBM. The activities of SOD and GPx were increased by PBM. The expression of Hsp1a/b (HSP70), a cell damage biomarker, was increased by hypoxia, and this effect was blocked by the action of PBM, demonstrating HSP70 levels similar to the control. Increased levels of TBARS and free radicals were found in the hypoxia group compared to the control, interestingly, in the group previously treated with pre-hypoxic PBM therapy we observed a decrease in these oxidative stress markers, bringing values similar to the control. Together, the data suggest that PBM has the potential as a cell protection therapy against oxidative damage under hypoxic conditions, evidenced by the expression of antioxidant enzymes and the reduction of levels of carbonylated proteins and free radicals.Considerado por muito tempo como camada celular inerte, o endotélio é reconhecido como um importante regulador das principais funções fisiológicas. As células endoteliais respondem a estímulos químicos e secretam fatores que modulam diversos processos fisiológicos, principalmente sob hipóxia. Essa condição pode desencadear respostas adaptativas e ter efeitos prejudiciais se persistir por períodos prolongados, causando modificações transcricionais que resultam em alterações da homeostase celular. A terapia de fotobiomodulação (FBM) pode estimular a produção de colágeno, promover a síntese de DNA e proteínas, aumentar o conteúdo de ATP, modular a migração e proliferação celular, acelera o reparo tecidual e induz a diferenciação celular. Nesse contexto, a avaliação das adaptações celulares à hipóxia pode fornecer conhecimentos para melhorar os tratamentos e reduzir essas perdas. O presente estudo in vitro teve como objetivo avaliar os efeitos da FBM pré indução de hipóxia em células endoteliais da veia umbilical humana (HUVECS), para prevenir possíveis danos gerados pela hipóxia. A metodologia compreendeu expressão gênica e atividade proteica através de testes in vitro. As células foram caracterizadas pela expressão de CD31 APC e CD146 APC pelo método de citometria de fluxo. As células, mantidas em cultura, foram soltas de uma garrafa matriz de 75cm2 e semeadas em garrafas de 25 cm² na quantidade 1x106. Nossos resultados evidenciaram que hipóxia apenas promoveu forte redução da viabilidade celular e que FBM com laser vermelho 660 nm; 1,5 J/cm2; 60s) pré hipóxia aumentou este parâmetro. As expressões de RNAm das enzimas antioxidantes SOD1 e CAT foram reduzidas pela hipóxia, e foram aumentadas com o uso da FBM. As atividades de SOD e GPx foram aumentadas pela FBM. A expressão de Hsp1a/b (HSP70), biomarcador de dano celular, mostrou-se aumentada pela hipóxia, sendo este efeito bloqueado pela ação da PBM, demostrando níveis de HSP70 semelhantes ao controle. Níveis aumentados de TBARS e de radicais livres foram encontrados no grupo hipóxia em comparação ao controle, interessantemente, no grupo previamente tratado com terapia de FBM pré hipóxia observamos queda destes marcadores de estresse oxidativo, trazendo valores semelhantes ao controle. Em conjunto, os dados sugerem que a FBM possui potencial como terapia de proteção celular em relação aos danos oxidativos em condições de hipóxia, evidenciado pela expressão de enzimas antioxidantes e redução dos níveis de proteínas carboniladas e radicais livres.Submitted by Nadir Basilio (nadirsb@uninove.br) on 2024-04-19T19:29:57Z No. of bitstreams: 1 Guilherme Rabelo Nasuk.pdf: 1233917 bytes, checksum: 54f718768dda3b01501c8a3657603bfe (MD5)Made available in DSpace on 2024-04-19T19:29:57Z (GMT). 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dc.title.por.fl_str_mv Efeito da terapia de fotobiomodulação na expressão de biomarcadores de estresse oxidativo em células HUVEC submetidas à hipóxia
dc.title.alternative.eng.fl_str_mv Effect of photobiomodulation therapy on the expression of oxidative stress biomarkers in HUVEC cells subjected to hypoxia
title Efeito da terapia de fotobiomodulação na expressão de biomarcadores de estresse oxidativo em células HUVEC submetidas à hipóxia
spellingShingle Efeito da terapia de fotobiomodulação na expressão de biomarcadores de estresse oxidativo em células HUVEC submetidas à hipóxia
Nasuk, Guilherme Rabelo
fotobiomodulação
HUVEC
enzimas antioxidantes
estresse oxidativo
photobiomodulation
HUVEC
antioxidant enzymes
oxidative stress
CIENCIAS DA SAUDE
title_short Efeito da terapia de fotobiomodulação na expressão de biomarcadores de estresse oxidativo em células HUVEC submetidas à hipóxia
title_full Efeito da terapia de fotobiomodulação na expressão de biomarcadores de estresse oxidativo em células HUVEC submetidas à hipóxia
title_fullStr Efeito da terapia de fotobiomodulação na expressão de biomarcadores de estresse oxidativo em células HUVEC submetidas à hipóxia
title_full_unstemmed Efeito da terapia de fotobiomodulação na expressão de biomarcadores de estresse oxidativo em células HUVEC submetidas à hipóxia
title_sort Efeito da terapia de fotobiomodulação na expressão de biomarcadores de estresse oxidativo em células HUVEC submetidas à hipóxia
author Nasuk, Guilherme Rabelo
author_facet Nasuk, Guilherme Rabelo
author_role author
dc.contributor.advisor1.fl_str_mv Silva Junior, Jose Antonio
dc.contributor.advisor1Lattes.fl_str_mv http://lattes.cnpq.br/1990427833073161
dc.contributor.referee1.fl_str_mv Silva Junior, Jose Antonio
dc.contributor.referee1Lattes.fl_str_mv http://lattes.cnpq.br/1990427833073161
dc.contributor.referee2.fl_str_mv Zamuner, Stella Regina
dc.contributor.referee2Lattes.fl_str_mv http://lattes.cnpq.br/1935805744318404
dc.contributor.referee3.fl_str_mv Ferrari, Raquel Agnelli Mesquita
dc.contributor.referee3Lattes.fl_str_mv http://lattes.cnpq.br/7048141683554943
dc.contributor.authorLattes.fl_str_mv http://lattes.cnpq.br/1816053013460118
dc.contributor.author.fl_str_mv Nasuk, Guilherme Rabelo
contributor_str_mv Silva Junior, Jose Antonio
Silva Junior, Jose Antonio
Zamuner, Stella Regina
Ferrari, Raquel Agnelli Mesquita
dc.subject.por.fl_str_mv fotobiomodulação
HUVEC
enzimas antioxidantes
estresse oxidativo
topic fotobiomodulação
HUVEC
enzimas antioxidantes
estresse oxidativo
photobiomodulation
HUVEC
antioxidant enzymes
oxidative stress
CIENCIAS DA SAUDE
dc.subject.eng.fl_str_mv photobiomodulation
HUVEC
antioxidant enzymes
oxidative stress
dc.subject.cnpq.fl_str_mv CIENCIAS DA SAUDE
description Long considered to be an inert cell layer, the endothelium is now recognized as an important regulator of major physiological functions. Endothelial cells respond to chemical stimuli and secrete factors that modulate various physiological processes, especially under hypoxia. This condition can trigger adaptive responses and have harmful effects if it persists for prolonged periods, causing transcriptional changes that result in changes in cellular homeostasis. Photobiomodulation therapy (PBM) can stimulate collagen production, promote DNA and protein synthesis, increase ATP content, modulate cell migration and proliferation, accelerate tissue repair, and induce cell differentiation. In this context, the evaluation of cellular adaptations to hypoxia can provide knowledge to improve treatments and reduce these losses. This in vitro study aimed to evaluate the effects of pre-hypoxic PBM on human umbilical vein endothelial cells (HUVECs), to prevent possible damage caused by hypoxia. The methodology included gene expression and protein activity through in vitro tests. The cells were characterized by the expression of CD31 APC and CD146 APC by flow cytometry. The cells, kept in culture, were released from a 75cm2 matrix bottle and seeded in 25 cm² bottles at a quantity of 1x106. Our results showed that hypoxia only promoted a strong reduction in cell viability and that PBM applied pre-hypoxia increased this parameter. The mRNA expressions of the antioxidant enzymes SOD1 and CAT were reduced by hypoxia, and were increased with the use of PBM. The activities of SOD and GPx were increased by PBM. The expression of Hsp1a/b (HSP70), a cell damage biomarker, was increased by hypoxia, and this effect was blocked by the action of PBM, demonstrating HSP70 levels similar to the control. Increased levels of TBARS and free radicals were found in the hypoxia group compared to the control, interestingly, in the group previously treated with pre-hypoxic PBM therapy we observed a decrease in these oxidative stress markers, bringing values similar to the control. Together, the data suggest that PBM has the potential as a cell protection therapy against oxidative damage under hypoxic conditions, evidenced by the expression of antioxidant enzymes and the reduction of levels of carbonylated proteins and free radicals.
publishDate 2023
dc.date.issued.fl_str_mv 2023-12-19
dc.date.accessioned.fl_str_mv 2024-04-19T19:29:57Z
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dc.identifier.citation.fl_str_mv Nasuk, Guilherme Rabelo. Efeito da terapia de fotobiomodulação na expressão de biomarcadores de estresse oxidativo em células HUVEC submetidas à hipóxia. 2023. 73 f. Dissertação( Programa de Mestrado em Medicina) - Universidade Nove de Julho, São Paulo.
dc.identifier.uri.fl_str_mv http://bibliotecatede.uninove.br/handle/tede/3345
identifier_str_mv Nasuk, Guilherme Rabelo. Efeito da terapia de fotobiomodulação na expressão de biomarcadores de estresse oxidativo em células HUVEC submetidas à hipóxia. 2023. 73 f. Dissertação( Programa de Mestrado em Medicina) - Universidade Nove de Julho, São Paulo.
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dc.publisher.country.fl_str_mv Brasil
dc.publisher.department.fl_str_mv Saúde
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