Efeito da fotobiomodulação sobre macrófagos submetidos à ação do veneno de B. jararacussu
Autor(a) principal: | |
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Data de Publicação: | 2020 |
Tipo de documento: | Tese |
Idioma: | por |
Título da fonte: | Biblioteca Digital de Teses e Dissertações da Uninove |
Texto Completo: | http://bibliotecatede.uninove.br/handle/tede/2757 |
Resumo: | Bothrops snake venom induces serious inflammatory events, including leukocyte migration and a complex network of mediators released. Leukocyte infiltration is characterized by an initial influx of neutrophils followed by a late infiltration of mononuclear cells, which accumulates at the injection site and in the surrounding tissues. Studies carried out by our group show that the treatment with photobiomodulation (PBM) associated with serotherapy decreases the leukocyte influx, in vivo, improving an inflammatory condition, but the mechanism by which photobiomolulation acts on the inflammatory reaction associated with the venom is unknown. Thus, the objective of this study was to evaluate the mechanism of action of PBM in RAW 264.7 macrophages incubated with botropic venom. For this, the cells were incubated with B. jararacussu venom (BjsuV), and immediately irradiated with Low level Laser with the following parameters: λ 660 nm wavelength, 100 mW power (power density 0.33 W / cm2), beam area 1 cm2, with an energy density of 4 J / cm2 and a time of 40 seconds, and were incubated at 1, 3, 6 and 24 h. The effect of PBM on cell viability was analyzed. In addition, the release of pro- and anti-inflammatory cytokines (IL-1, IL-6, TNF-α and IL-10) was analyzed by immunoenzymatic assay and intracellular calcium. The results obtained from the response curve showed no decrease in cell viability at any concentration used in the period of 1 h, in the period of 6 h all concentrations used demonstrated a decrease in cell viability. PBM caused a significant increase in cell viability, compared to the untreated group. The BjsuV group induced the release of IL-6, IL-1β with a significant difference at all period of time studied, with greater release at 3 h and TNF-α at 1 and 6 h. In addition, it showed a significant decreased in the release of IL-10, within 6 and 24 h. PBM decreased levels of TNF-α, at times 1 and 6 h, of IL-6, at times 3 and 6 h, at IL-1β, decreased levels at all times studied, and increased the release of IL -10 at 6 and 24 h. The BjsuV group showed a decrease in measurement of intracellular Ca2+, when compared to the control. PBM caused a significant increase in Ca2+. These data suggest that the protective effect of PBM on macrophages must be, at least in part, by decreasing pro-inflammatory cytokines and increasing anti-inflammatory. |
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Zamuner, Stella Reginahttp://lattes.cnpq.br/1935805744318404Zamuner, Stella Reginahttp://lattes.cnpq.br/1935805744318404Dalboni, Maria Aparecidahttp://lattes.cnpq.br/9818040147487320Correia, Marilia de Almeidahttp://lattes.cnpq.br/5983044080494563Gonçalves, Luis Roberto de Camargohttp://lattes.cnpq.br/8136459659811640http://lattes.cnpq.br/4733350256655516Campos, Gabriela Russo Soeiro2021-11-18T15:27:25Z2020-02-27Campos, Gabriela Russo Soeiro. Efeito da fotobiomodulação sobre macrófagos submetidos à ação do veneno de B. jararacussu. 2020. 40 f. Tese( Programa de Mestrado em Medicina) - Universidade Nove de Julho, São Paulo.http://bibliotecatede.uninove.br/handle/tede/2757Bothrops snake venom induces serious inflammatory events, including leukocyte migration and a complex network of mediators released. Leukocyte infiltration is characterized by an initial influx of neutrophils followed by a late infiltration of mononuclear cells, which accumulates at the injection site and in the surrounding tissues. Studies carried out by our group show that the treatment with photobiomodulation (PBM) associated with serotherapy decreases the leukocyte influx, in vivo, improving an inflammatory condition, but the mechanism by which photobiomolulation acts on the inflammatory reaction associated with the venom is unknown. Thus, the objective of this study was to evaluate the mechanism of action of PBM in RAW 264.7 macrophages incubated with botropic venom. For this, the cells were incubated with B. jararacussu venom (BjsuV), and immediately irradiated with Low level Laser with the following parameters: λ 660 nm wavelength, 100 mW power (power density 0.33 W / cm2), beam area 1 cm2, with an energy density of 4 J / cm2 and a time of 40 seconds, and were incubated at 1, 3, 6 and 24 h. The effect of PBM on cell viability was analyzed. In addition, the release of pro- and anti-inflammatory cytokines (IL-1, IL-6, TNF-α and IL-10) was analyzed by immunoenzymatic assay and intracellular calcium. The results obtained from the response curve showed no decrease in cell viability at any concentration used in the period of 1 h, in the period of 6 h all concentrations used demonstrated a decrease in cell viability. PBM caused a significant increase in cell viability, compared to the untreated group. The BjsuV group induced the release of IL-6, IL-1β with a significant difference at all period of time studied, with greater release at 3 h and TNF-α at 1 and 6 h. In addition, it showed a significant decreased in the release of IL-10, within 6 and 24 h. PBM decreased levels of TNF-α, at times 1 and 6 h, of IL-6, at times 3 and 6 h, at IL-1β, decreased levels at all times studied, and increased the release of IL -10 at 6 and 24 h. The BjsuV group showed a decrease in measurement of intracellular Ca2+, when compared to the control. PBM caused a significant increase in Ca2+. These data suggest that the protective effect of PBM on macrophages must be, at least in part, by decreasing pro-inflammatory cytokines and increasing anti-inflammatory.O veneno das serpentes do gênero Bothrops induz um conjunto de eventos inflamatórios graves, incluindo a migração de leucócitos e uma complexa rede de mediadores liberados no local. A infiltração de leucócitos é caracterizada por um influxo inicial de neutrófilos seguido por um infiltrado tardio de células mononucleares, que se acumula no local da injeção de veneno e nos tecidos adjacentes. Estudos realizados pelo nosso grupo, mostram que o tratamento com fotobiomodulação (FBM) associado a soroterapia diminui o influxo leucocitário, in vivo, melhorando todo um quadro inflamatório, porém ainda não se sabe o mecanismo pelo qual a fotobiomolulação age no quadro inflamatório associado ao veneno. Assim, o objetivo desse estudo foi avaliar o mecanismo de ação da FBM em macrófagos RAW 264.7 incubados com veneno botrópico. Para isso, as células foram incubadas com veneno de B. jararacussu (VBjsu), e imediatamente irradiadas com Laser de baixa intensidade com seguintes parâmetros: comprimento de onda de λ 660 nm, potência de 100 mW (densidade de potência de 0,33 W/cm2), área do feixe de 1 cm2, com densidade de energia de 4 J/cm2 e tempo de 40 segundos, e foram incubados nos tempos de 1, 3, 6 e 24 h. Foram analisados o efeito da fotobiomodulação sobre a viabilidade e integridade celular. Ainda, foi analisado a liberação de citocinas pró- e anti-inflamatórias (IL-1, IL-6, TNF-α e IL-10) por ensaio imunoenzimático e cálcio intracelular. Os resultados obtidos de curva dosem resposta não demonstraram diminuição da viabilidade celular em nenhuma concentração utilizada no período de 1 h, no período de 6 h todas as concentrações utilizadas demonstraram diminuição de viabilidade celular. A FBM causou aumento significante na viabilidade celular, comparado ao grupo sem tratamento. O grupo VBjsu induziu liberação de IL-6 e IL-1β com diferença significante em todos os tempos estudados, apresentando maior liberação no tempo de 3 h e de TNF- α no period de 1 e 6 h. Ainda, apresentou diminuição significante na liberação de IL-10, no tempo de 6 e 24 h. A FBM significantemente diminuiu os níveis de TNF-α, nos tempos de 1 e 6 h, e de IL-6, nos tempos de 3 e 6 h, para a citocina IL-1β, a FBM diminuiu os níveis em todos tempos estudados, e aumentou a liberação de IL-10 no tempo de 6 e 24 h. O grupo VBjsu causou diminuição de Ca2+ intracelular, quando comparado ao controle. A FBM causou uma elevação significativa do Ca2+. Esses dados sugerem que o efeito protetor da fotobiomodulação em macrófagos deve ser, pelo menos em parte, por diminuir as citocinas pró-inflamatórias e aumentar a anti-inflamatória.Submitted by Nadir Basilio (nadirsb@uninove.br) on 2021-11-18T15:27:25Z No. of bitstreams: 1 Gabriela Russo.pdf: 644686 bytes, checksum: c1c3fa9ff2e6fe996f6182db62ec4f6d (MD5)Made available in DSpace on 2021-11-18T15:27:25Z (GMT). No. of bitstreams: 1 Gabriela Russo.pdf: 644686 bytes, checksum: c1c3fa9ff2e6fe996f6182db62ec4f6d (MD5) Previous issue date: 2020-02-27application/pdfporUniversidade Nove de JulhoPrograma de Mestrado em MedicinaUNINOVEBrasilSaúdebothropscitocinasinflamaçãomacrófagosfotobiomodulaçãobothropscytokinesmacrophageinflammationphotobiomodulationCIENCIAS DA SAUDEEfeito da fotobiomodulação sobre macrófagos submetidos à ação do veneno de B. jararacussuEffect of photobiomodulation on macrophages subjected to the action of B. jararacussu venominfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/doctoralThesis8765449414823306929600info:eu-repo/semantics/openAccessreponame:Biblioteca Digital de Teses e Dissertações da Uninoveinstname:Universidade Nove de Julho (UNINOVE)instacron:UNINOVEORIGINALGabriela Russo.pdfGabriela Russo.pdfapplication/pdf644686http://localhost:8080/tede/bitstream/tede/2757/2/Gabriela+Russo.pdfc1c3fa9ff2e6fe996f6182db62ec4f6dMD52LICENSElicense.txtlicense.txttext/plain; charset=utf-82165http://localhost:8080/tede/bitstream/tede/2757/1/license.txtbd3efa91386c1718a7f26a329fdcb468MD51tede/27572021-11-18 13:27:25.106oai:localhost: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Biblioteca Digital de Teses e Dissertaçõeshttp://bibliotecatede.uninove.br/PRIhttp://bibliotecatede.uninove.br/oai/requestbibliotecatede@uninove.br||bibliotecatede@uninove.bropendoar:2021-11-18T15:27:25Biblioteca Digital de Teses e Dissertações da Uninove - Universidade Nove de Julho (UNINOVE)false |
dc.title.por.fl_str_mv |
Efeito da fotobiomodulação sobre macrófagos submetidos à ação do veneno de B. jararacussu |
dc.title.alternative.eng.fl_str_mv |
Effect of photobiomodulation on macrophages subjected to the action of B. jararacussu venom |
title |
Efeito da fotobiomodulação sobre macrófagos submetidos à ação do veneno de B. jararacussu |
spellingShingle |
Efeito da fotobiomodulação sobre macrófagos submetidos à ação do veneno de B. jararacussu Campos, Gabriela Russo Soeiro bothrops citocinas inflamação macrófagos fotobiomodulação bothrops cytokines macrophage inflammation photobiomodulation CIENCIAS DA SAUDE |
title_short |
Efeito da fotobiomodulação sobre macrófagos submetidos à ação do veneno de B. jararacussu |
title_full |
Efeito da fotobiomodulação sobre macrófagos submetidos à ação do veneno de B. jararacussu |
title_fullStr |
Efeito da fotobiomodulação sobre macrófagos submetidos à ação do veneno de B. jararacussu |
title_full_unstemmed |
Efeito da fotobiomodulação sobre macrófagos submetidos à ação do veneno de B. jararacussu |
title_sort |
Efeito da fotobiomodulação sobre macrófagos submetidos à ação do veneno de B. jararacussu |
author |
Campos, Gabriela Russo Soeiro |
author_facet |
Campos, Gabriela Russo Soeiro |
author_role |
author |
dc.contributor.advisor1.fl_str_mv |
Zamuner, Stella Regina |
dc.contributor.advisor1Lattes.fl_str_mv |
http://lattes.cnpq.br/1935805744318404 |
dc.contributor.referee1.fl_str_mv |
Zamuner, Stella Regina |
dc.contributor.referee1Lattes.fl_str_mv |
http://lattes.cnpq.br/1935805744318404 |
dc.contributor.referee2.fl_str_mv |
Dalboni, Maria Aparecida |
dc.contributor.referee2Lattes.fl_str_mv |
http://lattes.cnpq.br/9818040147487320 |
dc.contributor.referee3.fl_str_mv |
Correia, Marilia de Almeida |
dc.contributor.referee3Lattes.fl_str_mv |
http://lattes.cnpq.br/5983044080494563 |
dc.contributor.referee4.fl_str_mv |
Gonçalves, Luis Roberto de Camargo |
dc.contributor.referee4Lattes.fl_str_mv |
http://lattes.cnpq.br/8136459659811640 |
dc.contributor.authorLattes.fl_str_mv |
http://lattes.cnpq.br/4733350256655516 |
dc.contributor.author.fl_str_mv |
Campos, Gabriela Russo Soeiro |
contributor_str_mv |
Zamuner, Stella Regina Zamuner, Stella Regina Dalboni, Maria Aparecida Correia, Marilia de Almeida Gonçalves, Luis Roberto de Camargo |
dc.subject.por.fl_str_mv |
bothrops citocinas inflamação macrófagos fotobiomodulação |
topic |
bothrops citocinas inflamação macrófagos fotobiomodulação bothrops cytokines macrophage inflammation photobiomodulation CIENCIAS DA SAUDE |
dc.subject.eng.fl_str_mv |
bothrops cytokines macrophage inflammation photobiomodulation |
dc.subject.cnpq.fl_str_mv |
CIENCIAS DA SAUDE |
description |
Bothrops snake venom induces serious inflammatory events, including leukocyte migration and a complex network of mediators released. Leukocyte infiltration is characterized by an initial influx of neutrophils followed by a late infiltration of mononuclear cells, which accumulates at the injection site and in the surrounding tissues. Studies carried out by our group show that the treatment with photobiomodulation (PBM) associated with serotherapy decreases the leukocyte influx, in vivo, improving an inflammatory condition, but the mechanism by which photobiomolulation acts on the inflammatory reaction associated with the venom is unknown. Thus, the objective of this study was to evaluate the mechanism of action of PBM in RAW 264.7 macrophages incubated with botropic venom. For this, the cells were incubated with B. jararacussu venom (BjsuV), and immediately irradiated with Low level Laser with the following parameters: λ 660 nm wavelength, 100 mW power (power density 0.33 W / cm2), beam area 1 cm2, with an energy density of 4 J / cm2 and a time of 40 seconds, and were incubated at 1, 3, 6 and 24 h. The effect of PBM on cell viability was analyzed. In addition, the release of pro- and anti-inflammatory cytokines (IL-1, IL-6, TNF-α and IL-10) was analyzed by immunoenzymatic assay and intracellular calcium. The results obtained from the response curve showed no decrease in cell viability at any concentration used in the period of 1 h, in the period of 6 h all concentrations used demonstrated a decrease in cell viability. PBM caused a significant increase in cell viability, compared to the untreated group. The BjsuV group induced the release of IL-6, IL-1β with a significant difference at all period of time studied, with greater release at 3 h and TNF-α at 1 and 6 h. In addition, it showed a significant decreased in the release of IL-10, within 6 and 24 h. PBM decreased levels of TNF-α, at times 1 and 6 h, of IL-6, at times 3 and 6 h, at IL-1β, decreased levels at all times studied, and increased the release of IL -10 at 6 and 24 h. The BjsuV group showed a decrease in measurement of intracellular Ca2+, when compared to the control. PBM caused a significant increase in Ca2+. These data suggest that the protective effect of PBM on macrophages must be, at least in part, by decreasing pro-inflammatory cytokines and increasing anti-inflammatory. |
publishDate |
2020 |
dc.date.issued.fl_str_mv |
2020-02-27 |
dc.date.accessioned.fl_str_mv |
2021-11-18T15:27:25Z |
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dc.identifier.citation.fl_str_mv |
Campos, Gabriela Russo Soeiro. Efeito da fotobiomodulação sobre macrófagos submetidos à ação do veneno de B. jararacussu. 2020. 40 f. Tese( Programa de Mestrado em Medicina) - Universidade Nove de Julho, São Paulo. |
dc.identifier.uri.fl_str_mv |
http://bibliotecatede.uninove.br/handle/tede/2757 |
identifier_str_mv |
Campos, Gabriela Russo Soeiro. Efeito da fotobiomodulação sobre macrófagos submetidos à ação do veneno de B. jararacussu. 2020. 40 f. Tese( Programa de Mestrado em Medicina) - Universidade Nove de Julho, São Paulo. |
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http://bibliotecatede.uninove.br/handle/tede/2757 |
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