Dexamethasone lowers cytosolic pH in macrophages by altering alkalinizing pH-regulatory mechanisms.
The effect of dexamethasone on cytosolic pH (pHc) in resident mouse peritoneal macrophages was investigated using the fluorescent probe 2',7'-bis(carboxyethyl)-5(6)-carboxyfluorescein tetra-acetoxymethyl ester (BCECF-AM). Dexamethasone was found to significantly lower pHc and this reduction of pHc evolved gradually with time, was near maximal at 10 nM dexamethasone, and could be prevented by the glucocorticoid receptor antagonist RU-38486. The lower pHc of dexamethasone-treated cells was neither due to a reduction of cellular buffer capacity nor to an altered regulation of pHc by Na+/H+-exchange or by acidifying Na+-independent Cl-/HCO3- exchange, as assessed by studies of pH recovery after acute acid and alkali loads, respectively. Instead, an impaired pHc recovery by both the H+-ATPase and the alkalinizing Na+-dependent Cl-/HCO3- exchange was observed. This impairment was most likely not caused by an altered expression or localization of the 39-kDa subunit of the proton pump. Dexamethasone treatment caused a reduction of pHc also in a HCO3--containing solution, suggesting that acid extrusion by both the H+-ATPase and Na+-dependent Cl-/HCO3- exchange is important for maintenance and regulation of macrophage resting pHc. The lowering of macrophage pHc might be one mechanism whereby glucocorticoids exert their anti-inflammatory effects.
Nauclér
C
C
Department of Cell and Molecular Biology, Lund University, Sweden. claes.naucler@medkem.lu.se
Sundler
R
R
Tapper
H
H
D000136:Acid-Base Equilibrium / Q000187:drug effects*; D000143:Acids / Q000378:metabolism; D000468:Alkalies; D000818:Animals; D002021:Buffers; D003600:Cytosol / Q000187:drug effects; D003907:Dexamethasone / Q000378:metabolism / Q000494:pharmacology*; D006863:Hydrogen-Ion Concentration; D017737:Macrophages, Peritoneal / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D051379:Mice; D006180:Proton-Translocating ATPases / Q000378:metabolism
D000143:Acids; D000468:Alkalies; D002021:Buffers; D003907:Dexamethasone; D006180:Proton-Translocating ATPases