Avaliação do transporte transmembrânico de glicose, glutamina e alanil-glutamina em íleo de camundongos nutridos e desnutridos infectados por Escherichia coli enteropatogênica

Detalhes bibliográficos
Autor(a) principal: Sampaio, Ingrid Cordeiro
Data de Publicação: 2013
Tipo de documento: Dissertação
Idioma: por
Título da fonte: Repositório Institucional da Universidade Federal do Ceará (UFC)
Texto Completo: http://www.repositorio.ufc.br/handle/riufc/17971
Resumo: Enteropathogenic Escherichia coli (EPEC) and malnutrition are a major cause of morbidity and mortality in developing countries. The purpose of this study was to evaluate the transport of Na+-glucose, Na+-glutamine and H+-alanyl-glutamine by measurement of short circuit current (Isc) and transepithelial resistance (TR) in the ileum of nourished and malnourished mice infected with EPEC. Albino mice of approximately 35 days of age were subjected to a standard diet or a diet deficient in proteins, lipids and minerals, the Regional Basic Diet (RBD). Of these animals was submitted to acute infection for 3 hours of direct contact with bacteria in isolated intestinal loop. For comparison, the same was done using the commensal bacterium Escherichia coli HS. They were anesthetized with ketamine and xylazine intramuscularly. The ileum was removed, dissected the serous membrane, open with a scalpel, segmented and mounted in Üssing chambers. The total mRNA was extracted and, from this, cDNA was done to analyze if malnutrition caused alterations in gene transcription of transporters as SGLT-1, SN-2, CAT-1, PEPT-1 and CFTR and related proteins to cell junctions such as ZO-1, Claudin-1 and Claudin-2 and occludin. At the same time, immunofluorescence analysis of proteins SGLT-1, SN-1, SN-2 and PEPT-1 were performed. The results showed that EPEC reduced the area of the villi only during malnutrition (1,339 ± 0,1 mm²). But, the basal CCC did not alter (67,41 ± 2,0 μA/ cm²), while EPEC, E. coli HS and malnutrition, when isolated, reduced the basal CCC (65,96 ± 5,1 μA/ cm ², 72,51 ± 7,7 μA/ cm ² and 74,23 ± 5,7 μA/ cm²). However, malnutrition did not change the transport systems of glucose (ΔCCC = 324,7 ± 45,42 µA/cm²), glutamine (ΔCCC = 236.7 ± 43.32 μA/ cm²) and alanyl-glutamine (ΔCCC = 282,6 ± 39,10 μA/ cm²), because occured an increase in gene expression of SGLT-1 and PEPT-1 and increased synthesis of SGLT-1, SN-1, SN-2 and PEPT-1. Already EPEC altered glucose transport (ΔCCC = 102,6 ± 38,3 μA/ cm²), and E. coli HS decreased transport of glucose (ΔCCC = 103,5 ± 106,6 μA/ cm²) and glutamine (ΔCCC = 123,4 ± 106,5 μA/ cm²), whereas in undernourished animals EPEC reduced transport of glutamine (ΔCCC = 62,10 ± 42,7 μA/ cm²), and E. coli HS reduced levels of glucose absorption (ΔCCC = 84,40 ± 178,3 μA/ cm²). Therefore, these substrates may facilitate the recovery of the membrane and alleviate the vicious cycle of malnutrition, infection and diarrhea by increasing absorption (CCC) and the maintenance of normal intestinal permeability (Rt), but before using them for the treatment it is essential to evaluate the nutritional status of the host and the presence of pathogens in their intestinal epithelium.
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spelling Avaliação do transporte transmembrânico de glicose, glutamina e alanil-glutamina em íleo de camundongos nutridos e desnutridos infectados por Escherichia coli enteropatogênicaEvaluation of the transmembrane transport of glucose, glutamine and alanyl-glutamine in ileum of malnourished and infected mice by enteropathogenic Escherichia coli.DesnutriçãoEscherichia coli EnteropatogênicaAbsorção IntestinalEnteropathogenic Escherichia coli (EPEC) and malnutrition are a major cause of morbidity and mortality in developing countries. The purpose of this study was to evaluate the transport of Na+-glucose, Na+-glutamine and H+-alanyl-glutamine by measurement of short circuit current (Isc) and transepithelial resistance (TR) in the ileum of nourished and malnourished mice infected with EPEC. Albino mice of approximately 35 days of age were subjected to a standard diet or a diet deficient in proteins, lipids and minerals, the Regional Basic Diet (RBD). Of these animals was submitted to acute infection for 3 hours of direct contact with bacteria in isolated intestinal loop. For comparison, the same was done using the commensal bacterium Escherichia coli HS. They were anesthetized with ketamine and xylazine intramuscularly. The ileum was removed, dissected the serous membrane, open with a scalpel, segmented and mounted in Üssing chambers. The total mRNA was extracted and, from this, cDNA was done to analyze if malnutrition caused alterations in gene transcription of transporters as SGLT-1, SN-2, CAT-1, PEPT-1 and CFTR and related proteins to cell junctions such as ZO-1, Claudin-1 and Claudin-2 and occludin. At the same time, immunofluorescence analysis of proteins SGLT-1, SN-1, SN-2 and PEPT-1 were performed. The results showed that EPEC reduced the area of the villi only during malnutrition (1,339 ± 0,1 mm²). But, the basal CCC did not alter (67,41 ± 2,0 μA/ cm²), while EPEC, E. coli HS and malnutrition, when isolated, reduced the basal CCC (65,96 ± 5,1 μA/ cm ², 72,51 ± 7,7 μA/ cm ² and 74,23 ± 5,7 μA/ cm²). However, malnutrition did not change the transport systems of glucose (ΔCCC = 324,7 ± 45,42 µA/cm²), glutamine (ΔCCC = 236.7 ± 43.32 μA/ cm²) and alanyl-glutamine (ΔCCC = 282,6 ± 39,10 μA/ cm²), because occured an increase in gene expression of SGLT-1 and PEPT-1 and increased synthesis of SGLT-1, SN-1, SN-2 and PEPT-1. Already EPEC altered glucose transport (ΔCCC = 102,6 ± 38,3 μA/ cm²), and E. coli HS decreased transport of glucose (ΔCCC = 103,5 ± 106,6 μA/ cm²) and glutamine (ΔCCC = 123,4 ± 106,5 μA/ cm²), whereas in undernourished animals EPEC reduced transport of glutamine (ΔCCC = 62,10 ± 42,7 μA/ cm²), and E. coli HS reduced levels of glucose absorption (ΔCCC = 84,40 ± 178,3 μA/ cm²). Therefore, these substrates may facilitate the recovery of the membrane and alleviate the vicious cycle of malnutrition, infection and diarrhea by increasing absorption (CCC) and the maintenance of normal intestinal permeability (Rt), but before using them for the treatment it is essential to evaluate the nutritional status of the host and the presence of pathogens in their intestinal epithelium.A Escherichia coli enteropatogênica (EPEC) e a desnutrição são uma das maiores causadoras de morbidade e mortalidade infantil em países subdesenvolvidos. Este estudo propõe avaliar o transporte de Na+-glicose, Na+-glutamina e H+-alanil-glutamina através da medida da corrente de curto circuito (CCC) e da resistência transepitelial (Rt) no íleo de camundongos nutridos e desnutridos infectados com EPEC. Camundongos albinos de 28 a 35 dias de idade foram submetidos a uma dieta padrão ou a uma dieta multideficiente em proteínas, lipídeos e minerais, a DBR. Parte desses animais foi submetida à infecção aguda por 3 horas de contato direto com a bactéria em alça ligada. Para comparações, o mesmo foi feito utilizando a bactéria comensal Escherichia coli HS. Foram anestesiados com ketamina e xilazina por via intramuscular. O íleo foi removido, dissecado da membrana serosa, aberto com bisturi, segmentado e montado em câmaras de Üssing. O mRNA total foi extraído e, a partir deste, foi feito o cDNA a fim de observar, por PCR em tempo real, se a desnutrição causou alteração na transcrição dos genes SGLT-1, SN-2, CAT-1, PEPT-1, CFTR, ZO-1, Claudina-1 e Claudina-2 e Ocludina. Em paralelo, análise por microscopia confocal das proteínas SGLT-1, SN-1, SN-2 e PEPT-1 foi realizada. Os resultados mostraram que a EPEC reduziu a área dos vilos apenas durante a desnutrição (1,339 ± 0,1 mm²). Mesmo assim, não alterou a CCC basal (67,41 ± 2,0 µA/cm²), enquanto que a EPEC, a E. coli HS e a desnutrição, quando isoladas, reduziram a CCC basal (65,96 ± 5,1 µA/cm², 72,51 ± 7,7 µA/cm² e 74,23 ± 5,7 µA/cm²). Porém, a desnutrição não alterou os sistemas de transporte de glicose (ΔCCC = 324,7 ± 45,42 µA/cm²), glutamina (ΔCCC = 236,7 ± 43,32 µA/cm²) e alanil-glutamina (ΔCCC = 282,6 ± 39,10 µA/cm²) devido ao aumento na expressão da transcrição gênica de SGLT-1 e PEPT-1 e ao aumento da síntese de SGLT-1, SN-1, SN-2 e PEPT-1. Já a EPEC alterou o transporte de glicose (ΔCCC = 102,6 ± 38,3 µA/cm²), e a E. Coli HS diminuiu o transporte de glicose (ΔCCC = 103,5 ± 106,6 µA/cm²) e de glutamina (ΔCCC = 123,4 ± 106,5 µA/cm²), enquanto que, nos animais desnutridos, a EPEC reduziu o transporte de glutamina (ΔCCC = 62,10 ± 42,7 µA/cm²), e E. Coli HS reduziu os níveis de absorção de glicose (ΔCCC = 84,40 ± 178,3 µA/cm²). Portanto, é possível que esses substratos favoreçam a recuperação da membrana e aliviem o ciclo vicioso de desnutrição, infecção e diarreia através do aumento da absorção (CCC) e recuperação da normalidade da permeabilidade intestinal (Rt). Porém, deve-se avaliar o estado nutricional do hospedeiro e se há presença de patógenos associados, a fim de eleger o melhor substrato para cada caso.Lima, Aldo Ângelo Moreira LimaSampaio, Ingrid Cordeiro2016-06-27T13:12:14Z2016-06-27T13:12:14Z2013-08-29info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisapplication/pdfSAMPAIO, I. C. Avaliação do transporte transmembrânico de glicose, glutamina e alanil-glutamina em íleo de camundongos nutridos e desnutridos infectados por Escherichia coli enteropatogênica. 2013. 94 f. Dissertação (Mestrado em Microbiologia Médica) - Faculdade de Medicina, Universidade Federal do Ceará, Fortaleza, 2013.http://www.repositorio.ufc.br/handle/riufc/17971porreponame:Repositório Institucional da Universidade Federal do Ceará (UFC)instname:Universidade Federal do Ceará (UFC)instacron:UFCinfo:eu-repo/semantics/openAccess2022-03-24T19:54:49Zoai:repositorio.ufc.br:riufc/17971Repositório InstitucionalPUBhttp://www.repositorio.ufc.br/ri-oai/requestbu@ufc.br || repositorio@ufc.bropendoar:2024-09-11T18:20:14.818929Repositório Institucional da Universidade Federal do Ceará (UFC) - Universidade Federal do Ceará (UFC)false
dc.title.none.fl_str_mv Avaliação do transporte transmembrânico de glicose, glutamina e alanil-glutamina em íleo de camundongos nutridos e desnutridos infectados por Escherichia coli enteropatogênica
Evaluation of the transmembrane transport of glucose, glutamine and alanyl-glutamine in ileum of malnourished and infected mice by enteropathogenic Escherichia coli.
title Avaliação do transporte transmembrânico de glicose, glutamina e alanil-glutamina em íleo de camundongos nutridos e desnutridos infectados por Escherichia coli enteropatogênica
spellingShingle Avaliação do transporte transmembrânico de glicose, glutamina e alanil-glutamina em íleo de camundongos nutridos e desnutridos infectados por Escherichia coli enteropatogênica
Sampaio, Ingrid Cordeiro
Desnutrição
Escherichia coli Enteropatogênica
Absorção Intestinal
title_short Avaliação do transporte transmembrânico de glicose, glutamina e alanil-glutamina em íleo de camundongos nutridos e desnutridos infectados por Escherichia coli enteropatogênica
title_full Avaliação do transporte transmembrânico de glicose, glutamina e alanil-glutamina em íleo de camundongos nutridos e desnutridos infectados por Escherichia coli enteropatogênica
title_fullStr Avaliação do transporte transmembrânico de glicose, glutamina e alanil-glutamina em íleo de camundongos nutridos e desnutridos infectados por Escherichia coli enteropatogênica
title_full_unstemmed Avaliação do transporte transmembrânico de glicose, glutamina e alanil-glutamina em íleo de camundongos nutridos e desnutridos infectados por Escherichia coli enteropatogênica
title_sort Avaliação do transporte transmembrânico de glicose, glutamina e alanil-glutamina em íleo de camundongos nutridos e desnutridos infectados por Escherichia coli enteropatogênica
author Sampaio, Ingrid Cordeiro
author_facet Sampaio, Ingrid Cordeiro
author_role author
dc.contributor.none.fl_str_mv Lima, Aldo Ângelo Moreira Lima
dc.contributor.author.fl_str_mv Sampaio, Ingrid Cordeiro
dc.subject.por.fl_str_mv Desnutrição
Escherichia coli Enteropatogênica
Absorção Intestinal
topic Desnutrição
Escherichia coli Enteropatogênica
Absorção Intestinal
description Enteropathogenic Escherichia coli (EPEC) and malnutrition are a major cause of morbidity and mortality in developing countries. The purpose of this study was to evaluate the transport of Na+-glucose, Na+-glutamine and H+-alanyl-glutamine by measurement of short circuit current (Isc) and transepithelial resistance (TR) in the ileum of nourished and malnourished mice infected with EPEC. Albino mice of approximately 35 days of age were subjected to a standard diet or a diet deficient in proteins, lipids and minerals, the Regional Basic Diet (RBD). Of these animals was submitted to acute infection for 3 hours of direct contact with bacteria in isolated intestinal loop. For comparison, the same was done using the commensal bacterium Escherichia coli HS. They were anesthetized with ketamine and xylazine intramuscularly. The ileum was removed, dissected the serous membrane, open with a scalpel, segmented and mounted in Üssing chambers. The total mRNA was extracted and, from this, cDNA was done to analyze if malnutrition caused alterations in gene transcription of transporters as SGLT-1, SN-2, CAT-1, PEPT-1 and CFTR and related proteins to cell junctions such as ZO-1, Claudin-1 and Claudin-2 and occludin. At the same time, immunofluorescence analysis of proteins SGLT-1, SN-1, SN-2 and PEPT-1 were performed. The results showed that EPEC reduced the area of the villi only during malnutrition (1,339 ± 0,1 mm²). But, the basal CCC did not alter (67,41 ± 2,0 μA/ cm²), while EPEC, E. coli HS and malnutrition, when isolated, reduced the basal CCC (65,96 ± 5,1 μA/ cm ², 72,51 ± 7,7 μA/ cm ² and 74,23 ± 5,7 μA/ cm²). However, malnutrition did not change the transport systems of glucose (ΔCCC = 324,7 ± 45,42 µA/cm²), glutamine (ΔCCC = 236.7 ± 43.32 μA/ cm²) and alanyl-glutamine (ΔCCC = 282,6 ± 39,10 μA/ cm²), because occured an increase in gene expression of SGLT-1 and PEPT-1 and increased synthesis of SGLT-1, SN-1, SN-2 and PEPT-1. Already EPEC altered glucose transport (ΔCCC = 102,6 ± 38,3 μA/ cm²), and E. coli HS decreased transport of glucose (ΔCCC = 103,5 ± 106,6 μA/ cm²) and glutamine (ΔCCC = 123,4 ± 106,5 μA/ cm²), whereas in undernourished animals EPEC reduced transport of glutamine (ΔCCC = 62,10 ± 42,7 μA/ cm²), and E. coli HS reduced levels of glucose absorption (ΔCCC = 84,40 ± 178,3 μA/ cm²). Therefore, these substrates may facilitate the recovery of the membrane and alleviate the vicious cycle of malnutrition, infection and diarrhea by increasing absorption (CCC) and the maintenance of normal intestinal permeability (Rt), but before using them for the treatment it is essential to evaluate the nutritional status of the host and the presence of pathogens in their intestinal epithelium.
publishDate 2013
dc.date.none.fl_str_mv 2013-08-29
2016-06-27T13:12:14Z
2016-06-27T13:12:14Z
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 SAMPAIO, I. C. Avaliação do transporte transmembrânico de glicose, glutamina e alanil-glutamina em íleo de camundongos nutridos e desnutridos infectados por Escherichia coli enteropatogênica. 2013. 94 f. Dissertação (Mestrado em Microbiologia Médica) - Faculdade de Medicina, Universidade Federal do Ceará, Fortaleza, 2013.
http://www.repositorio.ufc.br/handle/riufc/17971
identifier_str_mv SAMPAIO, I. C. Avaliação do transporte transmembrânico de glicose, glutamina e alanil-glutamina em íleo de camundongos nutridos e desnutridos infectados por Escherichia coli enteropatogênica. 2013. 94 f. Dissertação (Mestrado em Microbiologia Médica) - Faculdade de Medicina, Universidade Federal do Ceará, Fortaleza, 2013.
url http://www.repositorio.ufc.br/handle/riufc/17971
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reponame_str Repositório Institucional da Universidade Federal do Ceará (UFC)
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