• JoomlaWorks Simple Image Rotator
  • JoomlaWorks Simple Image Rotator
  • JoomlaWorks Simple Image Rotator
  • JoomlaWorks Simple Image Rotator
  • JoomlaWorks Simple Image Rotator
  • JoomlaWorks Simple Image Rotator
  • JoomlaWorks Simple Image Rotator
  • JoomlaWorks Simple Image Rotator
  • JoomlaWorks Simple Image Rotator
  • JoomlaWorks Simple Image Rotator
 
  Bookmark and Share
 
 
Doctoral Thesis
DOI
10.11606/T.42.2012.tde-17092012-110833
Document
Author
Full name
Olivia Beloto da Silva
Institute/School/College
Knowledge Area
Date of Defense
Published
São Paulo, 2012
Supervisor
Committee
Souza, Maria Oliveira de (President)
Boim, Mirian Aparecida
Girardi, Adriana Castello Costa
Oliveira, Eduardo Rebelato Lopes de
Ribeiro, Helayne de Freitas
Title in Portuguese
A PKA modula a localização do SGLT1 e indiretamente a recuperação do pHi, no tratamento com alta concentração de glicose.
Keywords in Portuguese
Ativação enzimática
Concentração de enzima
Concentração de glicose
Glicose
Proteína cinase A
Proteínas da membrana
Abstract in Portuguese
Esse estudo avaliou o efeito da glicose sobre a atividade da PKA e sua interação com o SGLT1 e os trocadores Na+/H+ (NHE1 e NHE3). As células HEK-293 foram transfectadas com hSGLT1 wild type (WT) ou mutante (S418H) e tratadas 20 dias com DMEM contendo glicose 5 mM ou 25 mM. Foi avaliada a expressão de hSGLT1, hSGLT1+GFP, NHE1, NHE3 e PKA. Foi avaliada a expressão de hSGLT1+GFP e SGLT2 na membrana, com ou sem H-89, Manose, Sucrose e Filipina. Foi avaliada a velocidade de recuperação do pH intracelular (dpHi/dt), onde a solução de NH4Cl foi substituída por uma solução de glicose 5 ou 25 mM ou ambas as soluções na vigência de H-89 ou S3226. Esses dados indicam que o aumento do AMPc pela glicose altera a expressão e atividade dos SGLTs e NHEs. Nossos experimentos utilizando a transfecção do SGLT1 demonstraram que as células regulam a distribuição de SGLT1 e 2 na membrana, frente ao aumento extracelular desse substrato e que as vias ativadas pela glicose afetam a capacidade de recuperação do pH intracelular (pHi), através do NHE3.
Title in English
PKA modulates the SGLT1 distribution and the pHi recovery rate indirectly, in the high glucose concentration treatment.
Keywords in English
concentration of enzyme
Concentration of glucose
enzymatic activation
Glucose
membrane proteins
protein kinase A
Abstract in English
This study evaluated the effect of glucose on the PKA activity and its interaction with SGLT1 and the Na+/H+ exchanger (NHE1 and NHE3). The HEK-293 cells were transfected with hSGLT1 wild type (WT) or mutant (S418H) and treated 20 days with DMEM containing glucose 5 mM or 25 mM. The hSGLT1, hSGLT1+GFP, NHE1, NHE3 and PKA expression was analyzed. The surface hSGLT1+GFP and SGLT2 expression with or without H-89, Manose, Sucrose and Filipina was evaluated. The pHi recovery rate (dpHi/dt) was analyzed replacing the NH4Cl solution to glucose 5 or 25 mM solution or both with H-89 or S3226. These data indicate that the glucose increases the cAMP concentration and alters the expression and activity of SGLTs and NHEs. Our experiments using SGLT1 transfection demonstrated that, in the treatment with high glucose concentration, HEK-293 cells regulate the SGLT1 and 2 cellular distribution and that the pathways activated by glucose impair the pHi recovery rate, through the NHE3.
 
WARNING - Viewing this document is conditioned on your acceptance of the following terms of use:
This document is only for private use for research and teaching activities. Reproduction for commercial use is forbidden. This rights cover the whole data about this document as well as its contents. Any uses or copies of this document in whole or in part must include the author's name.
Publishing Date
2012-10-19
 
WARNING: Learn what derived works are clicking here.
All rights of the thesis/dissertation are from the authors
Centro de Informática de São Carlos
Digital Library of Theses and Dissertations of USP. Copyright © 2001-2020. All rights reserved.