Avaliação das alterações nucleares e cromatínicas em linfócitos do sangue periférico de portadores de diabetes

Detalhes bibliográficos
Autor(a) principal: Pereira, Vanessa Carla de Souza
Data de Publicação: 2017
Tipo de documento: Dissertação
Idioma: por
Título da fonte: Repositório Institucional da UFU
Texto Completo: https://repositorio.ufu.br/handle/123456789/26890
http://dx.doi.org/10.14393/ufu.di.2018.855
Resumo: Diabetes mellitus (DM) is considered an heterogenous group of metabolic disorders with hyperglycemia as a common event, because of defects in insulin secretion and/or action. Hyperglycemic condition resultant of DM can have a key role in changing cellular homeostasis, as a result of functional and structural alterations in several cellular components, being chromatin and example. Some changes in chromatin may be associated with changes in gene expression in immune cells as lymphocytes, which can lead to the dysfunction immune response on diabetes mellitus . Therefore, our aim was to screen for changes in chromatin organization of peripheral blood lymphocytes of both types 1 and 2 diabetic and non-diabetic subjects. In the present study, 20 men and women, with age range between 18 to 55 years old, have participated. Subjects were divided into three groups: Group 1, DM1 subjects, Group 2, DM2 subjects, and Group3, non-diabetic subjects or control group, paired with the other two groups by age and gender. Peripheral blood samples were collected from all subjects in the morning after an 8 hours fasting period, followed by dosage of glycaemia and glycated hemoglobin. Peripherical blood mononuclear cells from heparinized samples were isolated using the Ficoll method. Isolated cells were cytocentrifuged onto microscopy slides, fixed and Feulgen-stained. For evaluation of morphometric parameters of nuclear and chromatinic organization, digital images of stained nuclei were subjected to image analysis, and the results were correlated with results from blood dosages. Similar changes in chromatin organization were found in blood lymphocytes for both types of diabetes, including increased chromatin packaging and reorganization in diabetic subjects, but with a much more pronounced evidence of aneuploidy on lymphocytes of DM1 subjects. Glycated hemoglobin levels were positively correlated with blood glucose levels in both types of diabetes. However, for group1 the only statistical significant correlation found was between glycaemia, glycated hemoglobin and DNA content. For the group 2, glycaemia, even showing smaller values when compared with those for DM1, correlated well with all nuclear parameters, except changes in DNA content, which were more discrete for this group. According to present results, it is clear that both types of diabetes, as a consequence of hyperglycemia, are associated with important structural alterations in nuclei of human lymphocytes, which, we believe, may be causative agents of functional changes in this cell type, with consequent decline in immune functions of individuals.
id UFU_723d2cd3ef75c91941ceb14ce8d890ef
oai_identifier_str oai:repositorio.ufu.br:123456789/26890
network_acronym_str UFU
network_name_str Repositório Institucional da UFU
repository_id_str
spelling Avaliação das alterações nucleares e cromatínicas em linfócitos do sangue periférico de portadores de diabetesEvaluation of nuclear and chromatin changes in peripheral blood lymphocytes of patients with diabetesDiabetes mellitusDiabetes mellitusLinfócitosLymphocytesCromatinaChromatinNúcleo celularCell nucleusCitologiaCytologyDiabetesDiabetesCNPQ::CIENCIAS DA SAUDEDiabetes mellitus (DM) is considered an heterogenous group of metabolic disorders with hyperglycemia as a common event, because of defects in insulin secretion and/or action. Hyperglycemic condition resultant of DM can have a key role in changing cellular homeostasis, as a result of functional and structural alterations in several cellular components, being chromatin and example. Some changes in chromatin may be associated with changes in gene expression in immune cells as lymphocytes, which can lead to the dysfunction immune response on diabetes mellitus . Therefore, our aim was to screen for changes in chromatin organization of peripheral blood lymphocytes of both types 1 and 2 diabetic and non-diabetic subjects. In the present study, 20 men and women, with age range between 18 to 55 years old, have participated. Subjects were divided into three groups: Group 1, DM1 subjects, Group 2, DM2 subjects, and Group3, non-diabetic subjects or control group, paired with the other two groups by age and gender. Peripheral blood samples were collected from all subjects in the morning after an 8 hours fasting period, followed by dosage of glycaemia and glycated hemoglobin. Peripherical blood mononuclear cells from heparinized samples were isolated using the Ficoll method. Isolated cells were cytocentrifuged onto microscopy slides, fixed and Feulgen-stained. For evaluation of morphometric parameters of nuclear and chromatinic organization, digital images of stained nuclei were subjected to image analysis, and the results were correlated with results from blood dosages. Similar changes in chromatin organization were found in blood lymphocytes for both types of diabetes, including increased chromatin packaging and reorganization in diabetic subjects, but with a much more pronounced evidence of aneuploidy on lymphocytes of DM1 subjects. Glycated hemoglobin levels were positively correlated with blood glucose levels in both types of diabetes. However, for group1 the only statistical significant correlation found was between glycaemia, glycated hemoglobin and DNA content. For the group 2, glycaemia, even showing smaller values when compared with those for DM1, correlated well with all nuclear parameters, except changes in DNA content, which were more discrete for this group. According to present results, it is clear that both types of diabetes, as a consequence of hyperglycemia, are associated with important structural alterations in nuclei of human lymphocytes, which, we believe, may be causative agents of functional changes in this cell type, with consequent decline in immune functions of individuals.Dissertação (Mestrado)O diabetes mellitus (DM) é considerado um grupo heterogêneo de distúrbios metabólicos que apresentam em comum a hiperglicemia decorrente de defeitos na secreção ou ação da insulina, ou ainda quando ambos ocorrem. A condição hiperglicêmica oriunda da instalação do DM pode desempenhar um papel importante na alteração da homeostasia celular, como resultado de alterações funcionais e estruturais em vários componentes celulares, por exemplo, na cromatina. Algumas alterações cromatínicas podem estar diretamente relacionadas a mudanças na expressão gênica em células de defesa do organismo como os linfócitos, cuja disfunção pode ser relacionada à alteração da função imune no diabetes mellitus . Portanto, nosso objetivo foi pesquisar mudanças na organização da cromatina de linfócitos do sangue periférico de indivíduos portadores de diabetes 1 e 2, quando comparados a indivíduos não portadores. No presente estudo, foram incluídos 20 participantes, homens e mulheres, com idade entre 18 e 55 anos, sendo casos versus controles pareados por sexo e idade. Os participantes foram divididos em três grupos: Grupo 1, portadores de DM tipo 1, Grupo 2, portadores de DM tipo 2, e o Grupo Controle, não portadores de diabetes mellitus . Amostras de sangue periférico foram coletadas de todos os indivíduos pela manhã após um período de jejum de 8 horas, seguida de dosagem de glicemia e hemoglobina glicada. As células mononucleares das amostras heparinizadas de cada participante foram isoladas usando o método do Ficoll. As células isoladas foram citocentrugadas em lâminas para microscopia, fixadas e coradas pela reação de Feulgen. Para avaliação de parâmetros morfométricos de organização nuclear e da cromatina, imagens digitais de núcleos corados foram submetidas a análise de imagem e os resultados foram correlacionados com os resultados das dosagens bioquímicas. Alterações semelhantes na organização da cromatina foram encontradas em linfócitos para ambos os tipos de diabetes, incluindo o aumento da compactação e reorganização da cromatina, mas com evidências muito mais pronunciadas de aneuploidia em linfócitos de indivíduos com DM1. Os níveis de hemoglobina glicada foram correlacionados positivamente com os níveis de glicose no sangue em DM1 e DM2. No entanto, para o grupo 1, a única correlação estatisticament significativa encontrada foi entre glicemia, hemoglobina glicada e conteúdo de DNA. Para o grupo 2, a glicemia, mesmo mostrando valores menores quando comparados com aqueles para DM1, correlacionou-se bem com todos os parâmetros nucleares, exceto as alterações no conteúdo de DNA, que foram mais discretas para este grupo. Pelos resultados obtidos pode-se inferir que ambos os tipos de diabetes, pela condição hiperglicêmica induzida pelo DM, estão associados a importantes alterações estruturais nos núcleos de linfócitos humanos, as quais, acreditamos, possam ser a causa para alterações funcionais neste tipo celular, com consequente prejuízo nas funções imunes dos indivíduos.Universidade Federal de UberlândiaBrasilPrograma de Pós-graduação em Biologia Celular e Estrutural AplicadasMoraes, Alberto da Silvahttp://lattes.cnpq.br/7303349255103658Alvares-Saraiva, Anuska Marcelinohttp://lattes.cnpq.br/2148473770830932Beletti, Marcelo Emíliohttp://lattes.cnpq.br/1895300906751714Pereira, Vanessa Carla de Souza2019-09-03T19:40:12Z2019-09-03T19:40:12Z2017-05-31info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisapplication/pdfPEREIRA, Vanessa Carla de Souza. Avaliação das alterações nucleares e cromatínicas em linfócitos do sangue periférico de portadores de diabetes. 2017. 87 f. Dissertação (Mestrado em Biologia Celular e Estrutural Aplicadas) - Universidade Federal de Uberlândia, 2019. DOI http://dx.doi.org/10.14393/ufu.di.2018.855https://repositorio.ufu.br/handle/123456789/26890http://dx.doi.org/10.14393/ufu.di.2018.855porinfo:eu-repo/semantics/openAccessreponame:Repositório Institucional da UFUinstname:Universidade Federal de Uberlândia (UFU)instacron:UFU2019-09-04T06:06:56Zoai:repositorio.ufu.br:123456789/26890Repositório InstitucionalONGhttp://repositorio.ufu.br/oai/requestdiinf@dirbi.ufu.bropendoar:2019-09-04T06:06:56Repositório Institucional da UFU - Universidade Federal de Uberlândia (UFU)false
dc.title.none.fl_str_mv Avaliação das alterações nucleares e cromatínicas em linfócitos do sangue periférico de portadores de diabetes
Evaluation of nuclear and chromatin changes in peripheral blood lymphocytes of patients with diabetes
title Avaliação das alterações nucleares e cromatínicas em linfócitos do sangue periférico de portadores de diabetes
spellingShingle Avaliação das alterações nucleares e cromatínicas em linfócitos do sangue periférico de portadores de diabetes
Pereira, Vanessa Carla de Souza
Diabetes mellitus
Diabetes mellitus
Linfócitos
Lymphocytes
Cromatina
Chromatin
Núcleo celular
Cell nucleus
Citologia
Cytology
Diabetes
Diabetes
CNPQ::CIENCIAS DA SAUDE
title_short Avaliação das alterações nucleares e cromatínicas em linfócitos do sangue periférico de portadores de diabetes
title_full Avaliação das alterações nucleares e cromatínicas em linfócitos do sangue periférico de portadores de diabetes
title_fullStr Avaliação das alterações nucleares e cromatínicas em linfócitos do sangue periférico de portadores de diabetes
title_full_unstemmed Avaliação das alterações nucleares e cromatínicas em linfócitos do sangue periférico de portadores de diabetes
title_sort Avaliação das alterações nucleares e cromatínicas em linfócitos do sangue periférico de portadores de diabetes
author Pereira, Vanessa Carla de Souza
author_facet Pereira, Vanessa Carla de Souza
author_role author
dc.contributor.none.fl_str_mv Moraes, Alberto da Silva
http://lattes.cnpq.br/7303349255103658
Alvares-Saraiva, Anuska Marcelino
http://lattes.cnpq.br/2148473770830932
Beletti, Marcelo Emílio
http://lattes.cnpq.br/1895300906751714
dc.contributor.author.fl_str_mv Pereira, Vanessa Carla de Souza
dc.subject.por.fl_str_mv Diabetes mellitus
Diabetes mellitus
Linfócitos
Lymphocytes
Cromatina
Chromatin
Núcleo celular
Cell nucleus
Citologia
Cytology
Diabetes
Diabetes
CNPQ::CIENCIAS DA SAUDE
topic Diabetes mellitus
Diabetes mellitus
Linfócitos
Lymphocytes
Cromatina
Chromatin
Núcleo celular
Cell nucleus
Citologia
Cytology
Diabetes
Diabetes
CNPQ::CIENCIAS DA SAUDE
description Diabetes mellitus (DM) is considered an heterogenous group of metabolic disorders with hyperglycemia as a common event, because of defects in insulin secretion and/or action. Hyperglycemic condition resultant of DM can have a key role in changing cellular homeostasis, as a result of functional and structural alterations in several cellular components, being chromatin and example. Some changes in chromatin may be associated with changes in gene expression in immune cells as lymphocytes, which can lead to the dysfunction immune response on diabetes mellitus . Therefore, our aim was to screen for changes in chromatin organization of peripheral blood lymphocytes of both types 1 and 2 diabetic and non-diabetic subjects. In the present study, 20 men and women, with age range between 18 to 55 years old, have participated. Subjects were divided into three groups: Group 1, DM1 subjects, Group 2, DM2 subjects, and Group3, non-diabetic subjects or control group, paired with the other two groups by age and gender. Peripheral blood samples were collected from all subjects in the morning after an 8 hours fasting period, followed by dosage of glycaemia and glycated hemoglobin. Peripherical blood mononuclear cells from heparinized samples were isolated using the Ficoll method. Isolated cells were cytocentrifuged onto microscopy slides, fixed and Feulgen-stained. For evaluation of morphometric parameters of nuclear and chromatinic organization, digital images of stained nuclei were subjected to image analysis, and the results were correlated with results from blood dosages. Similar changes in chromatin organization were found in blood lymphocytes for both types of diabetes, including increased chromatin packaging and reorganization in diabetic subjects, but with a much more pronounced evidence of aneuploidy on lymphocytes of DM1 subjects. Glycated hemoglobin levels were positively correlated with blood glucose levels in both types of diabetes. However, for group1 the only statistical significant correlation found was between glycaemia, glycated hemoglobin and DNA content. For the group 2, glycaemia, even showing smaller values when compared with those for DM1, correlated well with all nuclear parameters, except changes in DNA content, which were more discrete for this group. According to present results, it is clear that both types of diabetes, as a consequence of hyperglycemia, are associated with important structural alterations in nuclei of human lymphocytes, which, we believe, may be causative agents of functional changes in this cell type, with consequent decline in immune functions of individuals.
publishDate 2017
dc.date.none.fl_str_mv 2017-05-31
2019-09-03T19:40:12Z
2019-09-03T19:40:12Z
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 PEREIRA, Vanessa Carla de Souza. Avaliação das alterações nucleares e cromatínicas em linfócitos do sangue periférico de portadores de diabetes. 2017. 87 f. Dissertação (Mestrado em Biologia Celular e Estrutural Aplicadas) - Universidade Federal de Uberlândia, 2019. DOI http://dx.doi.org/10.14393/ufu.di.2018.855
https://repositorio.ufu.br/handle/123456789/26890
http://dx.doi.org/10.14393/ufu.di.2018.855
identifier_str_mv PEREIRA, Vanessa Carla de Souza. Avaliação das alterações nucleares e cromatínicas em linfócitos do sangue periférico de portadores de diabetes. 2017. 87 f. Dissertação (Mestrado em Biologia Celular e Estrutural Aplicadas) - Universidade Federal de Uberlândia, 2019. DOI http://dx.doi.org/10.14393/ufu.di.2018.855
url https://repositorio.ufu.br/handle/123456789/26890
http://dx.doi.org/10.14393/ufu.di.2018.855
dc.language.iso.fl_str_mv por
language por
dc.rights.driver.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
dc.publisher.none.fl_str_mv Universidade Federal de Uberlândia
Brasil
Programa de Pós-graduação em Biologia Celular e Estrutural Aplicadas
publisher.none.fl_str_mv Universidade Federal de Uberlândia
Brasil
Programa de Pós-graduação em Biologia Celular e Estrutural Aplicadas
dc.source.none.fl_str_mv reponame:Repositório Institucional da UFU
instname:Universidade Federal de Uberlândia (UFU)
instacron:UFU
instname_str Universidade Federal de Uberlândia (UFU)
instacron_str UFU
institution UFU
reponame_str Repositório Institucional da UFU
collection Repositório Institucional da UFU
repository.name.fl_str_mv Repositório Institucional da UFU - Universidade Federal de Uberlândia (UFU)
repository.mail.fl_str_mv diinf@dirbi.ufu.br
_version_ 1813711314756829184