High glucose concentration stimulates NHE-1 activity in distal nephron cells: The role of the Mek/Erk1/2/p90 and p38MAPK signaling pathways

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 pH i recovery rate was monitored with HOE-694 (50 μM) and/or S3226 (10 μM), 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 NHE-3 proteins. Under control conditions, we observed a greater contribution of NHE-1 to pH i recovery relative to the other H + transporters. Acute high glucose treatment increased the HOE-694-sensitive pH i recovery rate and p-Erk1/2 and p90 RSK abundance. These parameters were reduced by PD-98059, a Mek inhibitor (1 μM). Chronic high glucose treatment also increased the HOE-694-sensitive pH i recovery rate and p-p38MAPK abundance. Both parameters were reduced by SB-203580, a p38MAPK inhibitor (10 μM). 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 © 2014 S. Karger AG, Basel.