Influence of selective phosphodiesterase inhibitors on human neutrophil functions and levels of cAMP and Cai.
Chromatographic analysis of 3',5'-cyclic nucleotide phosphodiesterase (PDE) isoenzymes in the cytosol of human neutrophils shows the predominant presence of PDE IV (cAMP specific) and PDE V (cGMP specific). PDE IV is characterized by (1) cAMP selectivity, (2) a KM for cAMP of 1.2 microM and (3) a typical rank order of IC50-values for PDE inhibitors: 0.13, 0.17, 47 and 9.5 microM for PDE IV selective rolipram, PDE III/IV selective zardaverine, PDE III selective motapizone and unselective 3-isobutyl-1-methylxanthine (IBMX), respectively. Functions of polymorphonuclear leukocytes (PMN) such as N-formylmethionyl-leucyl-phenylalanine (fMLP)-stimulated superoxide release and fMLP/thimerosal elicited leukotriene (LT) biosynthesis are inhibited by these PDE inhibitors with the same rank order and even lower IC50-values. Measurements of changes in cytosolic Cai in Fura-2 loaded PMN demonstrate a transient Cai increase after stimulation with 0.1 microM fMLP and an additional sustained elevation of Cai levels in the presence of thimerosal. PDE inhibitors suppress this sustained phase of Cai release with the same rank order of IC50-values as LT biosynthesis. The correlation between fMLP/thimerosal-induced LT biosynthesis and Cai levels reveal a Cai threshold of 150 nM for arachidonic acid metabolism. cAMP levels in PMN were elevated by PDE inhibitors alone by less than 2-fold. In the presence of fMLP however, cAMP was increased up to 10-fold and the efficacy of PDE inhibitors to increase cAMP paralleled their potency to inhibit PDE IV.(ABSTRACT TRUNCATED AT 250 WORDS)
Schudt
C
C
Department of Biochemistry, Byk Gulden Pharmaceuticals, Federal Republic of Germany.
Winder
S
S
Forderkunz
S
S
Hatzelmann
A
A
Ullrich
V
V
D001094:Arachidonate 5-Lipoxygenase / Q000378:metabolism; D002118:Calcium / Q000378:metabolism*; D000242:Cyclic AMP / Q000378:metabolism*; D006801:Humans; D007527:Isoenzymes; D015289:Leukotrienes / Q000096:biosynthesis; D009504:Neutrophils / Q000187:drug effects* / Q000378:metabolism; D010726:Phosphodiesterase Inhibitors / Q000494:pharmacology*
D007527:Isoenzymes; D015289:Leukotrienes; D010726:Phosphodiesterase Inhibitors; D000242:Cyclic AMP; D001094:Arachidonate 5-Lipoxygenase; D002118:Calcium