ESTUDO DO PAPEL DA HISTIDINA, HISTAMINA E ANTAGONISTAS HISTAMINÉRGICOS NA PROGRESSÃO E DIFERENCIAÇÃO CELULAR EM UM MODELO DE MELANOMA MURINO
Autor(a) principal: | |
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Data de Publicação: | 2015 |
Tipo de documento: | Dissertação |
Idioma: | por |
Título da fonte: | Biblioteca Digital de Teses e Dissertações da UEPG |
Texto Completo: | http://tede2.uepg.br/jspui/handle/prefix/91 |
Resumo: | Histidine is an important amino acid in the differentiation of some cell types, such as hepatocytes, for example. Histamine is synthesized from histidine is present in many cancer types, including melanoma. This cancer has as characteristics no differentiated cell and the production a high amount of histamine. The purpose of this work was to study the effects of a histidine supplementation on differentiation and tumor progression. In vitro cell viability were performed by neutral red and MTT assays with histidine at 0,01, 0,1, 0,3, 1,0, 3,0, 10 and 20 mM cimetidine at 0,01, 0,1, 0,3, 1,0, 3,0, 10 mM, 10 mM histidine association cimetidine + 0,01 mM and 10 mM cimetidine association + 0,01 mM histidine and evaluated after 24, 48 and 72 hours using B16F10 cells and morphological analysis additionally performed. Histamine at 0,01, 0,1, 1 and 10 mM, terfenadine at concentrations 1, 10, 100 nM and 1 uM and thioperamide at concentrations 1, 10, 100 nM and 1 uM were also assessed by using the MTT cell B16F10. ELISA was performed to quantify histamine with cell culture supernatant treated with histidine, histamine, terfenadine, cimetidine, and thioperamide. For in vivo assays were used C57BL6/J mice underwent implantation of tumor cells and subsequently treated with histidine solution at a dose of 21.4 mg/kg cimetidine 343.3 mg/kg and association between both and evaluated for 11 days as the tumor growth were used. Histological analysis, hematological and biochemical determinations were also performed. Data were analysed by ANOVA One-Way, and Tukey post-test. The results of the MTT viability test, showed cell viability decreased histidine at a concentration of 0,01 mM in 72 hours, cimetidine in concentrations 3 to 10 mM at 48 and 72 hours and cimetidine 10 mM /histidine 0,01 mM association in 48 and 72 hours, results also found in neutral red. Histamine decreased the cell viability of B16F10 cells at 20 mM concentration in 48 h and concentrations 1 and 10 mM at 72 hours, terfenadine decreased cell viability at all concentrations tested at 24 h and stayed up to 10 nM 72 hours and thioperamide decreased the cell viability at all concentrations tested 72 hours MTT assay. In the morphological analysis it was observed that at lower concentrations of histidine discrete cell rounding occurred and the appearance of "blebs" and the highest concentrations of vacuolated cytoplasm. Cimetidine treated cells showed characteristics of apoptosis. The association histidine 0,01 mM/cimetidine 10 mM caused cell death. The histamine ELISA test showed that the treatment with the histamine H1 antagonist may increase or decrease production of histamine and its receiving treatment led to thioperamide produce less or blocking of histamine H3 receptors allows a greater uptake of histamine. In experiments in vivo treatments with histidine and cimetidine inhibited tumor growth by the seventh day of treatment. Biochemical determinations of AST, ALT showed no toxicity after treatment. The histological findings, the group treated with histidine showed necrotic areas and the cimetidine group showed leukocyte infiltration and promoted stimulation of angiogenesis. Metastases were not found in any of the organs analyzed groups. It was concluded that histidine plays an important role in tumor progression and stimulating or inhibitory depending on the dosage/day of treatment. |
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Faveró, Giovani MarinoCPF:02640460994http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4770965A4Franco, Claudinéia Conationi da SilvaCPF:03680727976http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4596571U0Justus, José Fabiano CostaCPF:01710741902http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4118424U9CPF:02621108903http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4778518J5Berton, Juliana2017-07-21T14:13:04Z2015-10-162017-07-21T14:13:04Z2015-02-26BERTON, Juliana. ESTUDO DO PAPEL DA HISTIDINA, HISTAMINA E ANTAGONISTAS HISTAMINÉRGICOS NA PROGRESSÃO E DIFERENCIAÇÃO CELULAR EM UM MODELO DE MELANOMA MURINO. 2015. 90 f. Dissertação (Mestrado em Farmacos, Medicamentos e Biociências Aplicadas à Farmácia) - UNIVERSIDADE ESTADUAL DE PONTA GROSSA, Ponta Grossa, 2015.http://tede2.uepg.br/jspui/handle/prefix/91Histidine is an important amino acid in the differentiation of some cell types, such as hepatocytes, for example. Histamine is synthesized from histidine is present in many cancer types, including melanoma. This cancer has as characteristics no differentiated cell and the production a high amount of histamine. The purpose of this work was to study the effects of a histidine supplementation on differentiation and tumor progression. In vitro cell viability were performed by neutral red and MTT assays with histidine at 0,01, 0,1, 0,3, 1,0, 3,0, 10 and 20 mM cimetidine at 0,01, 0,1, 0,3, 1,0, 3,0, 10 mM, 10 mM histidine association cimetidine + 0,01 mM and 10 mM cimetidine association + 0,01 mM histidine and evaluated after 24, 48 and 72 hours using B16F10 cells and morphological analysis additionally performed. Histamine at 0,01, 0,1, 1 and 10 mM, terfenadine at concentrations 1, 10, 100 nM and 1 uM and thioperamide at concentrations 1, 10, 100 nM and 1 uM were also assessed by using the MTT cell B16F10. ELISA was performed to quantify histamine with cell culture supernatant treated with histidine, histamine, terfenadine, cimetidine, and thioperamide. For in vivo assays were used C57BL6/J mice underwent implantation of tumor cells and subsequently treated with histidine solution at a dose of 21.4 mg/kg cimetidine 343.3 mg/kg and association between both and evaluated for 11 days as the tumor growth were used. Histological analysis, hematological and biochemical determinations were also performed. Data were analysed by ANOVA One-Way, and Tukey post-test. The results of the MTT viability test, showed cell viability decreased histidine at a concentration of 0,01 mM in 72 hours, cimetidine in concentrations 3 to 10 mM at 48 and 72 hours and cimetidine 10 mM /histidine 0,01 mM association in 48 and 72 hours, results also found in neutral red. Histamine decreased the cell viability of B16F10 cells at 20 mM concentration in 48 h and concentrations 1 and 10 mM at 72 hours, terfenadine decreased cell viability at all concentrations tested at 24 h and stayed up to 10 nM 72 hours and thioperamide decreased the cell viability at all concentrations tested 72 hours MTT assay. In the morphological analysis it was observed that at lower concentrations of histidine discrete cell rounding occurred and the appearance of "blebs" and the highest concentrations of vacuolated cytoplasm. Cimetidine treated cells showed characteristics of apoptosis. The association histidine 0,01 mM/cimetidine 10 mM caused cell death. The histamine ELISA test showed that the treatment with the histamine H1 antagonist may increase or decrease production of histamine and its receiving treatment led to thioperamide produce less or blocking of histamine H3 receptors allows a greater uptake of histamine. In experiments in vivo treatments with histidine and cimetidine inhibited tumor growth by the seventh day of treatment. Biochemical determinations of AST, ALT showed no toxicity after treatment. The histological findings, the group treated with histidine showed necrotic areas and the cimetidine group showed leukocyte infiltration and promoted stimulation of angiogenesis. Metastases were not found in any of the organs analyzed groups. It was concluded that histidine plays an important role in tumor progression and stimulating or inhibitory depending on the dosage/day of treatment.A histidina é um aminoácido importante fator na diferenciação de alguns tipos celulares, como hepatócitos, por exemplo. A histamina, sintetizada a partir da histidina, está presente em muitos tipos de câncer, entre eles o melanoma. Esse tipo de câncer tem como características a não diferenciação celular e a produção de grande quantidade de histamina. O propósito deste trabalho foi estudar os efeitos de uma suplementação de histidina sobre a diferenciação e progressão tumoral. Os ensaios in vitro de viabilidade celular MTT e vermelho neutro foram realizados com histidina nas concentrações 0,01, 0,1, 0,3, 1,0, 3,0, 10 e 20 mM, cimetidina nas concentrações 0,01, 0,1, 0,3, 1,0, 3,0, 10 mM, associação histidina 10 mM + cimetidina 0,01 mM e associação de cimetidina 10 mM + histidina 0,01 mM e avaliados após 24, 48 e 72 horas utilizando-se células B16F10 e adicionalmente realizada a análise morfológica. Histamina nas concentrações 0,01, 0,1, 1 e 10 mM, terfenadina nas concentrações 1, 10, 100 nM e 1 μM e tioperamida nas concentrações 1, 10, 100 nM e 1 μM também foram avaliadas pelo MTT utilizandose células B16F10. Foi realizado teste de ELISA para quantificação de histamina com sobrenadante de cultura celular tratada com histidina, histamina, terfenadina, cimetidina e tioperamida. Para o ensaio in vivo foram utilizados camundongos C57BL6/J, submetidos ao implante de células tumorais e posteriormente tratados com solução de histidina na dose de 21,4 mg/kg, cimetidina na dose de 343,3 mg/kg e associação entre ambas e avaliados por 11 dias quanto ao crescimento tumoral. Avaliações hematológicas, determinações bioquímicas e análise histológica também foram realizadas. Os dados obtidos foram submetidos à análise estatística ANOVA 1-VIA, e pos-test Tukey. Como resultados no teste de viabilidade MTT, a histidina diminuiu a viabilidade celular na concentração de 0,01 mM em 72 horas, da cimetidina nas concentrações de 3 e 10 mm em 48 e 72 horas e da associação cimetidina 10 mM/histidina 0,01 mM em 48 e 72 horas, resultados também encontrados no vermelho neutro. A histamina diminuiu a viabilidade celular das células B16F10 em 48 h na concentração de 20 mM e em 72 horas nas concentrações de 1 e 10 mM, a terfenadina diminuiu a viabilidade celular em todas as concentrações testadas em 24 h, permanecendo a de 10 nM até 72 horas e a tioperamida diminuiu a viabilidade celular em todas as concentrações testadas em 72 horas no ensaio de MTT. Na análise morfológica observou-se que nas menores concentrações de histidina ocorreu discreto arredondamento celular e aparecimento de “blebs” e nas concentrações maiores vacuolização de citoplasma. As células tratadas com cimetidina apresentaram indícios de apoptose. A associação histidina 0,01 mM/cimetidina 10 mM provocou morte celular. O teste de ELISA para histamina revelou que o tratamento com o antagonista H1 de histamina pode aumentar a produção de histamina ou diminuir sua receptação e o tratamento a tioperamida levou a uma menor produção de histamina ou o bloqueio dos receptores H3 permitindo uma maior recaptação de histamina. Na experimentação in vivo os tratamentos com histidina e cimetidina inibiram o crescimento tumoral até o sétimo dia de tratamento. As determinações bioquímicas das enzimas transaminase oxalacética e transaminase pirúvica revelaram que não houve toxicidade após os tratamentos. Nas avaliações histológicas, o grupo tratado com histidina apresentou áreas necróticas e o grupo cimetidina apresentou infiltrado leucocitário e promoveu estímulo da angiogênese. Metástases não foram encontradas nos órgãos analisados de nenhum dos grupos. Concluiu-se que a histidina tem um papel importante na progressão tumoral sendo inibitório ou estimulante dependendo da dosagem/dias de tratamento.Made available in DSpace on 2017-07-21T14:13:04Z (GMT). No. of bitstreams: 1 Berton, Juliana.pdf: 2602704 bytes, checksum: 2cb56990fbaa5b2f82264280c97a04df (MD5) Previous issue date: 2015-02-26Coordenação de Aperfeiçoamento de Pessoal de Nível Superiorapplication/pdfporUNIVERSIDADE ESTADUAL DE PONTA GROSSAPrograma de Pós Graduação Ciências FarmacêuticasUEPGBRFarmacos, Medicamentos e Biociências Aplicadas à FarmáciamelanomahistidinaB16F10MelanomaHistidineB16F10CNPQ::CIENCIAS DA SAUDE::FARMACIAESTUDO DO PAPEL DA HISTIDINA, HISTAMINA E ANTAGONISTAS HISTAMINÉRGICOS NA PROGRESSÃO E DIFERENCIAÇÃO CELULAR EM UM MODELO DE MELANOMA MURINOinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisinfo:eu-repo/semantics/openAccessreponame:Biblioteca Digital de Teses e Dissertações da UEPGinstname:Universidade Estadual de Ponta Grossa (UEPG)instacron:UEPGORIGINALBerton, Juliana.pdfapplication/pdf2602704http://tede2.uepg.br/jspui/bitstream/prefix/91/1/Berton%2c%20Juliana.pdf2cb56990fbaa5b2f82264280c97a04dfMD51prefix/912017-07-21 11:13:04.383oai:tede2.uepg.br:prefix/91Biblioteca Digital de Teses e Dissertaçõeshttps://tede2.uepg.br/jspui/PUBhttp://tede2.uepg.br/oai/requestbicen@uepg.br||mv_fidelis@yahoo.com.bropendoar:2017-07-21T14:13:04Biblioteca Digital de Teses e Dissertações da UEPG - Universidade Estadual de Ponta Grossa (UEPG)false |
dc.title.por.fl_str_mv |
ESTUDO DO PAPEL DA HISTIDINA, HISTAMINA E ANTAGONISTAS HISTAMINÉRGICOS NA PROGRESSÃO E DIFERENCIAÇÃO CELULAR EM UM MODELO DE MELANOMA MURINO |
title |
ESTUDO DO PAPEL DA HISTIDINA, HISTAMINA E ANTAGONISTAS HISTAMINÉRGICOS NA PROGRESSÃO E DIFERENCIAÇÃO CELULAR EM UM MODELO DE MELANOMA MURINO |
spellingShingle |
ESTUDO DO PAPEL DA HISTIDINA, HISTAMINA E ANTAGONISTAS HISTAMINÉRGICOS NA PROGRESSÃO E DIFERENCIAÇÃO CELULAR EM UM MODELO DE MELANOMA MURINO Berton, Juliana melanoma histidina B16F10 Melanoma Histidine B16F10 CNPQ::CIENCIAS DA SAUDE::FARMACIA |
title_short |
ESTUDO DO PAPEL DA HISTIDINA, HISTAMINA E ANTAGONISTAS HISTAMINÉRGICOS NA PROGRESSÃO E DIFERENCIAÇÃO CELULAR EM UM MODELO DE MELANOMA MURINO |
title_full |
ESTUDO DO PAPEL DA HISTIDINA, HISTAMINA E ANTAGONISTAS HISTAMINÉRGICOS NA PROGRESSÃO E DIFERENCIAÇÃO CELULAR EM UM MODELO DE MELANOMA MURINO |
title_fullStr |
ESTUDO DO PAPEL DA HISTIDINA, HISTAMINA E ANTAGONISTAS HISTAMINÉRGICOS NA PROGRESSÃO E DIFERENCIAÇÃO CELULAR EM UM MODELO DE MELANOMA MURINO |
title_full_unstemmed |
ESTUDO DO PAPEL DA HISTIDINA, HISTAMINA E ANTAGONISTAS HISTAMINÉRGICOS NA PROGRESSÃO E DIFERENCIAÇÃO CELULAR EM UM MODELO DE MELANOMA MURINO |
title_sort |
ESTUDO DO PAPEL DA HISTIDINA, HISTAMINA E ANTAGONISTAS HISTAMINÉRGICOS NA PROGRESSÃO E DIFERENCIAÇÃO CELULAR EM UM MODELO DE MELANOMA MURINO |
author |
Berton, Juliana |
author_facet |
Berton, Juliana |
author_role |
author |
dc.contributor.advisor1.fl_str_mv |
Faveró, Giovani Marino |
dc.contributor.advisor1ID.fl_str_mv |
CPF:02640460994 |
dc.contributor.advisor1Lattes.fl_str_mv |
http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4770965A4 |
dc.contributor.referee1.fl_str_mv |
Franco, Claudinéia Conationi da Silva |
dc.contributor.referee1ID.fl_str_mv |
CPF:03680727976 |
dc.contributor.referee1Lattes.fl_str_mv |
http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4596571U0 |
dc.contributor.referee2.fl_str_mv |
Justus, José Fabiano Costa |
dc.contributor.referee2ID.fl_str_mv |
CPF:01710741902 |
dc.contributor.referee2Lattes.fl_str_mv |
http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4118424U9 |
dc.contributor.authorID.fl_str_mv |
CPF:02621108903 |
dc.contributor.authorLattes.fl_str_mv |
http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4778518J5 |
dc.contributor.author.fl_str_mv |
Berton, Juliana |
contributor_str_mv |
Faveró, Giovani Marino Franco, Claudinéia Conationi da Silva Justus, José Fabiano Costa |
dc.subject.por.fl_str_mv |
melanoma histidina B16F10 |
topic |
melanoma histidina B16F10 Melanoma Histidine B16F10 CNPQ::CIENCIAS DA SAUDE::FARMACIA |
dc.subject.eng.fl_str_mv |
Melanoma Histidine B16F10 |
dc.subject.cnpq.fl_str_mv |
CNPQ::CIENCIAS DA SAUDE::FARMACIA |
description |
Histidine is an important amino acid in the differentiation of some cell types, such as hepatocytes, for example. Histamine is synthesized from histidine is present in many cancer types, including melanoma. This cancer has as characteristics no differentiated cell and the production a high amount of histamine. The purpose of this work was to study the effects of a histidine supplementation on differentiation and tumor progression. In vitro cell viability were performed by neutral red and MTT assays with histidine at 0,01, 0,1, 0,3, 1,0, 3,0, 10 and 20 mM cimetidine at 0,01, 0,1, 0,3, 1,0, 3,0, 10 mM, 10 mM histidine association cimetidine + 0,01 mM and 10 mM cimetidine association + 0,01 mM histidine and evaluated after 24, 48 and 72 hours using B16F10 cells and morphological analysis additionally performed. Histamine at 0,01, 0,1, 1 and 10 mM, terfenadine at concentrations 1, 10, 100 nM and 1 uM and thioperamide at concentrations 1, 10, 100 nM and 1 uM were also assessed by using the MTT cell B16F10. ELISA was performed to quantify histamine with cell culture supernatant treated with histidine, histamine, terfenadine, cimetidine, and thioperamide. For in vivo assays were used C57BL6/J mice underwent implantation of tumor cells and subsequently treated with histidine solution at a dose of 21.4 mg/kg cimetidine 343.3 mg/kg and association between both and evaluated for 11 days as the tumor growth were used. Histological analysis, hematological and biochemical determinations were also performed. Data were analysed by ANOVA One-Way, and Tukey post-test. The results of the MTT viability test, showed cell viability decreased histidine at a concentration of 0,01 mM in 72 hours, cimetidine in concentrations 3 to 10 mM at 48 and 72 hours and cimetidine 10 mM /histidine 0,01 mM association in 48 and 72 hours, results also found in neutral red. Histamine decreased the cell viability of B16F10 cells at 20 mM concentration in 48 h and concentrations 1 and 10 mM at 72 hours, terfenadine decreased cell viability at all concentrations tested at 24 h and stayed up to 10 nM 72 hours and thioperamide decreased the cell viability at all concentrations tested 72 hours MTT assay. In the morphological analysis it was observed that at lower concentrations of histidine discrete cell rounding occurred and the appearance of "blebs" and the highest concentrations of vacuolated cytoplasm. Cimetidine treated cells showed characteristics of apoptosis. The association histidine 0,01 mM/cimetidine 10 mM caused cell death. The histamine ELISA test showed that the treatment with the histamine H1 antagonist may increase or decrease production of histamine and its receiving treatment led to thioperamide produce less or blocking of histamine H3 receptors allows a greater uptake of histamine. In experiments in vivo treatments with histidine and cimetidine inhibited tumor growth by the seventh day of treatment. Biochemical determinations of AST, ALT showed no toxicity after treatment. The histological findings, the group treated with histidine showed necrotic areas and the cimetidine group showed leukocyte infiltration and promoted stimulation of angiogenesis. Metastases were not found in any of the organs analyzed groups. It was concluded that histidine plays an important role in tumor progression and stimulating or inhibitory depending on the dosage/day of treatment. |
publishDate |
2015 |
dc.date.available.fl_str_mv |
2015-10-16 2017-07-21T14:13:04Z |
dc.date.issued.fl_str_mv |
2015-02-26 |
dc.date.accessioned.fl_str_mv |
2017-07-21T14:13:04Z |
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info:eu-repo/semantics/publishedVersion |
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info:eu-repo/semantics/masterThesis |
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masterThesis |
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dc.identifier.citation.fl_str_mv |
BERTON, Juliana. ESTUDO DO PAPEL DA HISTIDINA, HISTAMINA E ANTAGONISTAS HISTAMINÉRGICOS NA PROGRESSÃO E DIFERENCIAÇÃO CELULAR EM UM MODELO DE MELANOMA MURINO. 2015. 90 f. Dissertação (Mestrado em Farmacos, Medicamentos e Biociências Aplicadas à Farmácia) - UNIVERSIDADE ESTADUAL DE PONTA GROSSA, Ponta Grossa, 2015. |
dc.identifier.uri.fl_str_mv |
http://tede2.uepg.br/jspui/handle/prefix/91 |
identifier_str_mv |
BERTON, Juliana. ESTUDO DO PAPEL DA HISTIDINA, HISTAMINA E ANTAGONISTAS HISTAMINÉRGICOS NA PROGRESSÃO E DIFERENCIAÇÃO CELULAR EM UM MODELO DE MELANOMA MURINO. 2015. 90 f. Dissertação (Mestrado em Farmacos, Medicamentos e Biociências Aplicadas à Farmácia) - UNIVERSIDADE ESTADUAL DE PONTA GROSSA, Ponta Grossa, 2015. |
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UNIVERSIDADE ESTADUAL DE PONTA GROSSA |
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Programa de Pós Graduação Ciências Farmacêuticas |
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UEPG |
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BR |
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Farmacos, Medicamentos e Biociências Aplicadas à Farmácia |
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UNIVERSIDADE ESTADUAL DE PONTA GROSSA |
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