Efeitos da leptina e SAA na função de neutrófilos humanos
Autor(a) principal: | |
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Data de Publicação: | 2020 |
Tipo de documento: | Dissertação |
Idioma: | por |
Título da fonte: | Repositório do Centro Universitário Braz Cubas |
Texto Completo: | https://repositorio.cruzeirodosul.edu.br/handle/123456789/2666 |
Resumo: | Although there is a consensus that the infections incidence is higher in obese, the effect of adipokines on neutrophil function is not completely known. Herein, we hypothesized that leptin and serum amyloid A (SAA) can activate/inactivate neutrophils and interfere with the innate immune response. So, we studied the combined effects of leptin and SAA on the function of human neutrophils. Neutrophils (2.5x106 cells / mL) were isolated and cultured in RPMI medium in the presence and absence of leptin (25 and 50ng/mL), SAA (5ug/mL) and LPS (1ug/mL). In vitro, we analyzed, through flow cytometry technique, the effect of leptin and SAA on parameters related to neutrophil viability; by ELISA we evaluated the release of cytokines (TNF-α, IL-8) and by the chemiluminescence technique amplified by lucigenin we observed the production of ROS. Under the conditions studied, neutrophil treatments with SAA, leptin and the combination SAA and leptin were not toxic to the cells. We observed an increase in ROS production of 60% (*p= 0.01) and 79% (**p= 0.02) by neutrophils, when treated with the combination of SAA and leptin at concentrations of 2.5ng/mL and 25ng/mL respectively. The treatment of neutrophils with leptin at concentrations of 25ng/mL, 50ng/mL and SAA increased by 38% (**p= 0.02) and 92% (***p= 0.003) and 159% (****p= 0.0002) the concentration of TNF-α and these cells, when treated, with leptin at concentrations of 25ng/mL and 50ng/mL with SAA showed an increase of 62% (°p= 0.0044), and 106% (°°°p= 0.0002), respectively. The treatment of neutrophils with leptin at concentrations 25ng / mL and 50ng/mL and SAA increased by 39% (**p= 0.01), 38% (***p= 0.01) and 62% (****p= 0.001) the concentration of IL-8, respectively, and these neutrophils when treated with leptin at a concentration of 50ng / mL with SAA showed an increase of 32% (°p= 0.0001). There was an increase in the production of TNF-α and IL-8 at different concentrations of leptin and in treatments with combined leptin and SAA. Our data indicate that adipose tissue can modulate neutrophil function through adipokine secretion and interfere with the innate immune response. |
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Efeitos da leptina e SAA na função de neutrófilos humanosEffects of leptin and SAA on function of human neutrophilsLeucócitosObesidadeInflamação.CNPQ::CIENCIAS DA SAUDEAlthough there is a consensus that the infections incidence is higher in obese, the effect of adipokines on neutrophil function is not completely known. Herein, we hypothesized that leptin and serum amyloid A (SAA) can activate/inactivate neutrophils and interfere with the innate immune response. So, we studied the combined effects of leptin and SAA on the function of human neutrophils. Neutrophils (2.5x106 cells / mL) were isolated and cultured in RPMI medium in the presence and absence of leptin (25 and 50ng/mL), SAA (5ug/mL) and LPS (1ug/mL). In vitro, we analyzed, through flow cytometry technique, the effect of leptin and SAA on parameters related to neutrophil viability; by ELISA we evaluated the release of cytokines (TNF-α, IL-8) and by the chemiluminescence technique amplified by lucigenin we observed the production of ROS. Under the conditions studied, neutrophil treatments with SAA, leptin and the combination SAA and leptin were not toxic to the cells. We observed an increase in ROS production of 60% (*p= 0.01) and 79% (**p= 0.02) by neutrophils, when treated with the combination of SAA and leptin at concentrations of 2.5ng/mL and 25ng/mL respectively. The treatment of neutrophils with leptin at concentrations of 25ng/mL, 50ng/mL and SAA increased by 38% (**p= 0.02) and 92% (***p= 0.003) and 159% (****p= 0.0002) the concentration of TNF-α and these cells, when treated, with leptin at concentrations of 25ng/mL and 50ng/mL with SAA showed an increase of 62% (°p= 0.0044), and 106% (°°°p= 0.0002), respectively. The treatment of neutrophils with leptin at concentrations 25ng / mL and 50ng/mL and SAA increased by 39% (**p= 0.01), 38% (***p= 0.01) and 62% (****p= 0.001) the concentration of IL-8, respectively, and these neutrophils when treated with leptin at a concentration of 50ng / mL with SAA showed an increase of 32% (°p= 0.0001). There was an increase in the production of TNF-α and IL-8 at different concentrations of leptin and in treatments with combined leptin and SAA. Our data indicate that adipose tissue can modulate neutrophil function through adipokine secretion and interfere with the innate immune response.Embora seja consenso que a incidência e o agravamento de infeções são maiores em obesos, o efeito de adipocinas, mediadores secretados pelo tecido adiposo, como a amilóide sérica A (SAA) e a leptina, na função de neutrófilos, primeira linha de defesa do organismo, não é completamente conhecido. Acreditando na hipótese de que a leptina e a SAA possam ativar/inativar neutrófilos e interferir na resposta imune inata, nesse trabalho estudamos os efeitos da leptina e de leptina e SAA na função de neutrófilos humanos. Neutrófilos (2,5x106 células/mL) foram isolados do sangue periférico de doadores saudáveis, e posteriormente, cultivados em meio RPMI na presença de leptina (25 e 50ng/mL), SAA (5ug/mL) e leptina nas diferentes concentrações em conjunto com a SAA. LPS (1ug/mL) foi utilizado como controle positivo. In vitro analisamos, através da técnica de citometria de fluxo o efeito da leptina e de SAA sobre parâmetros relacionados à viabilidade de neutrófilos; por ELISA avaliamos a liberação de TNF-α e IL-8 e pela técnica de quimiluminescência amplificada por lucigenina observamos a produção de ROS. Nas condições estudadas, os tratamentos de neutrófilos com SAA, leptina e a combinação de ambas não foram tóxicos para as células. Observamos aumento na produção de ROS de 60% (*p=0.01) e 79% (**p=0.02) por neutrófilos, quando tratados com a combinação de SAA e leptina nas concentrações de 2.5ng/mL e 25ng/mL respectivamente. O tratamento de neutrófilos com leptina nas concentrações de 25ng/mL, 50ng/mL e SAA isoladamente aumentou 38% (**p=0.02), 92% (***p=0.003) e 159% (****p=0.0002) a concentração de TNF-α, respectivamente e essas células, quando tratadas com leptina nas concentrações de 25ng/mL e 50ng/mL em conjunto com a SAA apresentaram aumento de 62% (°p=0.0044), e 106% (°°°p=0.0002), respectivamente. A concentração de IL-8 foi aumentada em neutrófilos tradados com leptina nas concentrações 25ng/mL e 50ng/mL e SAA isoladamente, 39% (**p=0.01), 38% (***p= 0.01) e 62% (****p=0.001), respectivamente e essas células quando tratados com leptina na concentração 50ng/mL em conjunto com a SAA apresentaram aumento de 32% (°p=0.0001). Os resultados indicam que a combinação de leptina e SAA induz a produção de ROS, TNF-α e IL-8 por neutrófilos. Os nossos dados apontam que o tecido adiposo pode através da secreção de adipocinas modular a função de neutrófilos e interferir na resposta imune inata.Universidade Cruzeiro do SulBrasilPrograma de Pós Graduação em Ciências da SaúdeCruzeiro do SulDermargos, Elaine Hatanaka18316791838http://lattes.cnpq.br/0256403741529064Dermargos, Elaine Hatanaka18316791838http://lattes.cnpq.br/0256403741529064Gorjão , Renatahttp://lattes.cnpq.br/6211339830408691Sandri, Silvanahttp://lattes.cnpq.br/3978430910797029Silva, Bianca Palmeira Santos da2021-08-20T16:58:30Z2020-05-072021-08-20T16:58:30Z2020info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisapplication/pdfPALMEIRA B. Efeitos da leptina e da SAA na função de neutrófilos humano. 58 f. 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Positional cloning of the mouse obese gene and its human homologue. Nature. 1994, n. 372, p. 425 432. ZHAO, S. et al. Partial leptin deficiency confers resistance to diet-induced obesity in mice. Mol. Metabol., 2020, n. 100995. ZICK, Y. Role of Ser/Thr kinases in the uncoupling of insulin signaling. Int. J. Obes. Relat. Metab. Disord, 2003, n. 27 (3), p. 56 60. ZYCHLINSKY, A. et al. Introduction: Forum in immunology on neutrophils. Microbes Infect., 2003, n. 5, p. 1289-1291.info:eu-repo/semantics/openAccessreponame:Repositório do Centro Universitário Braz Cubasinstname:Centro Universitário Braz Cubas (CUB)instacron:CUB2021-08-20T17:03:43Zoai:repositorio.cruzeirodosul.edu.br:123456789/2666Repositório InstitucionalPUBhttps://repositorio.brazcubas.edu.br/oai/requestbibli@brazcubas.edu.bropendoar:2021-08-20T17:03:43Repositório do Centro Universitário Braz Cubas - Centro Universitário Braz Cubas (CUB)false |
dc.title.none.fl_str_mv |
Efeitos da leptina e SAA na função de neutrófilos humanos Effects of leptin and SAA on function of human neutrophils |
title |
Efeitos da leptina e SAA na função de neutrófilos humanos |
spellingShingle |
Efeitos da leptina e SAA na função de neutrófilos humanos Silva, Bianca Palmeira Santos da Leucócitos Obesidade Inflamação. CNPQ::CIENCIAS DA SAUDE |
title_short |
Efeitos da leptina e SAA na função de neutrófilos humanos |
title_full |
Efeitos da leptina e SAA na função de neutrófilos humanos |
title_fullStr |
Efeitos da leptina e SAA na função de neutrófilos humanos |
title_full_unstemmed |
Efeitos da leptina e SAA na função de neutrófilos humanos |
title_sort |
Efeitos da leptina e SAA na função de neutrófilos humanos |
author |
Silva, Bianca Palmeira Santos da |
author_facet |
Silva, Bianca Palmeira Santos da |
author_role |
author |
dc.contributor.none.fl_str_mv |
Dermargos, Elaine Hatanaka 18316791838 http://lattes.cnpq.br/0256403741529064 Dermargos, Elaine Hatanaka 18316791838 http://lattes.cnpq.br/0256403741529064 Gorjão , Renata http://lattes.cnpq.br/6211339830408691 Sandri, Silvana http://lattes.cnpq.br/3978430910797029 |
dc.contributor.author.fl_str_mv |
Silva, Bianca Palmeira Santos da |
dc.subject.por.fl_str_mv |
Leucócitos Obesidade Inflamação. CNPQ::CIENCIAS DA SAUDE |
topic |
Leucócitos Obesidade Inflamação. CNPQ::CIENCIAS DA SAUDE |
description |
Although there is a consensus that the infections incidence is higher in obese, the effect of adipokines on neutrophil function is not completely known. Herein, we hypothesized that leptin and serum amyloid A (SAA) can activate/inactivate neutrophils and interfere with the innate immune response. So, we studied the combined effects of leptin and SAA on the function of human neutrophils. Neutrophils (2.5x106 cells / mL) were isolated and cultured in RPMI medium in the presence and absence of leptin (25 and 50ng/mL), SAA (5ug/mL) and LPS (1ug/mL). In vitro, we analyzed, through flow cytometry technique, the effect of leptin and SAA on parameters related to neutrophil viability; by ELISA we evaluated the release of cytokines (TNF-α, IL-8) and by the chemiluminescence technique amplified by lucigenin we observed the production of ROS. Under the conditions studied, neutrophil treatments with SAA, leptin and the combination SAA and leptin were not toxic to the cells. We observed an increase in ROS production of 60% (*p= 0.01) and 79% (**p= 0.02) by neutrophils, when treated with the combination of SAA and leptin at concentrations of 2.5ng/mL and 25ng/mL respectively. The treatment of neutrophils with leptin at concentrations of 25ng/mL, 50ng/mL and SAA increased by 38% (**p= 0.02) and 92% (***p= 0.003) and 159% (****p= 0.0002) the concentration of TNF-α and these cells, when treated, with leptin at concentrations of 25ng/mL and 50ng/mL with SAA showed an increase of 62% (°p= 0.0044), and 106% (°°°p= 0.0002), respectively. The treatment of neutrophils with leptin at concentrations 25ng / mL and 50ng/mL and SAA increased by 39% (**p= 0.01), 38% (***p= 0.01) and 62% (****p= 0.001) the concentration of IL-8, respectively, and these neutrophils when treated with leptin at a concentration of 50ng / mL with SAA showed an increase of 32% (°p= 0.0001). There was an increase in the production of TNF-α and IL-8 at different concentrations of leptin and in treatments with combined leptin and SAA. Our data indicate that adipose tissue can modulate neutrophil function through adipokine secretion and interfere with the innate immune response. |
publishDate |
2020 |
dc.date.none.fl_str_mv |
2020-05-07 2020 2021-08-20T16:58:30Z 2021-08-20T16:58:30Z |
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 |
PALMEIRA B. Efeitos da leptina e da SAA na função de neutrófilos humano. 58 f. Dissertação (Mestrado em Ciências da Saúde) - Universidade Cruzeiro do Sul, São Paulo, 2020. https://repositorio.cruzeirodosul.edu.br/handle/123456789/2666 |
identifier_str_mv |
PALMEIRA B. Efeitos da leptina e da SAA na função de neutrófilos humano. 58 f. Dissertação (Mestrado em Ciências da Saúde) - Universidade Cruzeiro do Sul, São Paulo, 2020. |
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https://repositorio.cruzeirodosul.edu.br/handle/123456789/2666 |
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por |
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Universidade Cruzeiro do Sul Brasil Programa de Pós Graduação em Ciências da Saúde Cruzeiro do Sul |
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