| Full text | |
| Author(s): |
da Costa-Pessoa, Juliana Martins
[1]
;
Damasceno, Roselia Santos
[1]
;
Machado, Ubiratan Fabres
[1]
;
Beloto-Silva, Olivia
[1]
;
Oliveira-Souza, Maria
[1]
Total Authors: 5
|
| Affiliation: | [1] Univ Sao Paulo, Inst Biomed Sci, Dept Physiol & Biophys, BR-05508900 Sao Paulo - Brazil
Total Affiliations: 1
|
| Document type: | Journal article |
| Source: | CELLULAR PHYSIOLOGY AND BIOCHEMISTRY; v. 33, n. 2, p. 333-343, 2014. |
| Web of Science Citations: | 8 |
| Abstract | |
Aims: In models of diabetes, distal nephron cells contribute to glucose uptake and oxidation. How these cells contribute to the use of glucose for the regulation of H+ extrusion remains unknown. We used Madin-Darby Canine Kidney (MDCK) cells to investigate the effect of acute or chronic high glucose concentration on the abundance and activity of the Na+/H+ exchanger (NHE-1). Methods: Using RT-PCR, we also evaluated the mRNA expression for sodium glucose co-transporters SGLT1 and SGLT2. Protein abundance was analyzed using immunoblotting, and intracellular pH (pH(i)) recovery was evaluated using microscopy in conjunction with the fluorescent probe BCECF/AM. The Na+-dependent pHi recovery rate was monitored with HOE-694 (50 mu M) and/or S3226 (10 mu M), specific NHE-1 and NHE-3 inhibitors. Results: MDCK cells did not express the mRNA for SGLT1 or SGLT2 but did express the GLUT2, NHE-1 and NHE3 proteins. Under control conditions, we observed a greater contribution of NHE-1 to pHi recovery relative to the other H+ transporters. Acute high glucose treatment increased the HOE-694- sensitive pHi recovery rate and p-Erk1/2 and p90(RSK) abundance. These parameters were reduced by PD-98059, a Mek inhibitor (1 mu M). Chronic high glucose treatment also increased the HOE-694-sensitive pHi recovery rate and p-p38MAPK abundance. Both parameters were reduced by SB-203580, a p38MAPK inhibitor (10 mu M). Conclusion: These results suggested that extracellular high glucose stimulated NHE-1 acutely and chronically through Mek/Erk1/2/ p90(RSK) and p38MAPK pathways, respectively. Copyright (C) 2014 S. Karger AG, Basel (AU) | |
| FAPESP's process: | 12/04831-1 - New players in glycemic control and chronic complications of Diabetes mellitus: preventive and therapeutic perspectives |
| Grantee: | Ubiratan Fabres Machado |
| Support Opportunities: | Research Projects - Thematic Grants |
| FAPESP's process: | 07/58966-7 - Analise da regulacao do ph intracelular em celulas renais. |
| Grantee: | Maria Oliveira de Souza |
| Support Opportunities: | Regular Research Grants |