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Forschungsdatenbank PMU-SQQUID

Effect of inhibitors of Na+/H+-exchange and gastric H+/K+ ATPase on cell volume, intracellular pH and migration of human polymorphonuclear leucocytes.
Ritter, M; Schratzberger, P; Rossmann, H; Wöll, E; Seiler, K; Seidler, U; Reinisch, N; Kähler, CM; Zwierzina, H; Lang, HJ; Lang, F; Paulmichl, M; Wiedermann, CJ;
Br J Pharmacol. 1998; 124(4): 627-638.
Originalarbeiten (Zeitschrift)

PMU-Autor/inn/en

Paulmichl Markus
Ritter Markus

Abstract

1. Stimulation of chemotaxis of human polymorphonuclear leucocytes (PMNs) with the chemoattractive peptide fMLP (N-formyl-Met-Leu-Phe) is paralleled by profound morphological and metabolic alterations like changes of intracellular pH (pHi) and cell shape. The present study was performed to investigate the interrelation of cell volume (CV) regulatory ion transport, pHi and migration of fMLP stimulated PMNs. 2. Addition of fMLP to PMNs stimulated directed migration in Boyden chamber assays and was accompanied by rapid initial intracellular acidification and cell swelling. 3. Inhibition of the Na+/H+ exchanger suppressed fMLP stimulated cell migration, accelerated the intracellular acidification and inhibited the fMLP-induced cell swelling. 4. Step omission of extracellular Na+ caused intracellular acidification, which was accelerated by subsequent addition of gastric H+/K+ ATPase inhibitor SCH 28080, or by omission of extracellular K+ ions. In addition Na+ removal caused cell swelling, which was further enhanced by fMLP. 5. H+/K+ATPase inhibitors omeprazole and SCH 28080 inhibited stimulated migration and blunted the fMLP-induced increase in CV. 6. Increasing extracellular osmolarity by addition of mannitol to the extracellular solution caused cell shrinkage followed by regulatory volume increase, partially due to activation of the Na+/H+ exchanger. In fMLP-stimulated cells the CV increase was counteracted by simultaneous addition of mannitol. Under these conditions the fMLP stimulated migration was inhibited. 7. The antibacterial activity of PMNs was not modified by Hoe 694 or omeprazole. 8. Western analysis with a monoclonal anti gastric H+/K+ ATPase beta-subunit antibody detected a glycosylated 35 kD core protein in lysates of mouse and human gastric mucosa as well as in human PMNs. 9. The results indicate that fMLP leads to cell swelling of PMNs due to activation of the Na+/H+ exchanger and a K+-dependent H+-extruding mechanism, presumably an H+/K+ ATPase. Inhibition of these ion transporters suppresses the increase in CV and precludes PMNs from stimulated migration.


Useful keywords (using NLM MeSH Indexing)

Animals

Blood Bactericidal Activity/drug effects

Blotting, Western

Cell Size/drug effects

Chemotaxis, Leukocyte/drug effects*

Chemotaxis, Leukocyte/physiology

Colony Count, Microbial

Enzyme Inhibitors/pharmacology

Escherichia coli/growth*

development

Gastric Mucosa/cytology

Gastric Mucosa/enzymology*

Gastric Mucosa/metabolism

H(+)-K(+)-Exchanging ATPase/antagonists*

inhibitors*

Humans

Hydrogen-Ion Concentration

Intracellular Fluid/chemistry

Ion Transport/drug effects

Mice

Mice, Inbred C57BL

Microscopy, Confocal

N-Formylmethionine Leucyl-Phenylalanine/pharmacology

Neutrophils/cytology

Neutrophils/drug effects*

Neutrophils/physiology

Sodium-Hydrogen Antiporter/antagonists*

inhibitors*


Find related publications in this database (Keywords)

leucocyte
cell migration
chemotaxis
cell volume
intracellular pH
Na+/H+ exchanger
Hoe694
Hoe642
K+/H+ ATPase
omeprazole
SCH28080