Inhibition of phosphodiesterase IV and intracellular calcium levels in human polymorphonuclear leukocytes.
Phosphodiesterase (PDE) isoenzyme type IV is the predominant cyclic AMP hydrolytic activity in polymorphonuclear leukocytes (PMNs). PDE IV inhibitors depress functional responses of PMNs but their influence on intracellular calcium concentration ([Ca2+]i) has not been extensively studied. The present study examined the effects of rolipram (a selective PDE IV inhibitor) on the chemotactic peptide formyl-methionyl-leucyl-phenylalanine (fMLP)-induced changes of [Ca2+]i in fura-2 loaded human PMNs. Rolipram (1 nM-10 microM) did not alter basal [Ca2+]i values. fMLP (10 nM approximately EC50) produced a transient calcium response, i.e., a peak followed by decay to a residual value above baseline. Peak [Ca2+]i values after fMLP were not altered but a faster decay and a lower residual [Ca2+]i were observed in rolipram (0.1-10 microM)-treated cells. fMLP added after thimerosal (20 microM) produced a peak followed by a sustained oscillatory response. Rolipram (up to 10 microM) did not alter the peak but inhibited the sustained response (-log IC50 = 6.39 +/- 0.12). The inhibitory effects of rolipram may be due to alterations in the mobilization of Ca2+ produced by the increase in the cellular content of cyclic AMP. SKF94120 (a selective PDE III inhibitor) produced minor effects on the fMLP-induced calcium response. SCA40 (a mixed PDE III/IV/V inhibitor) produced similar effects but was less potent than rolipram. Reduction of the calcium response probably underlies the inhibition of PMN functions produced by PDE IV inhibitors.
Villagrasa
V
V
Department of Pharmacology, Faculty of Medicine and Odontology, University of Valencia, Spain.
Navarrete
C
C
Sanz
C
C
Berto
L
L
Perpiñá
M
M
Cortijo
J
J
Morcillo
E J
EJ
D015105:3',5'-Cyclic-AMP Phosphodiesterases; D002118:Calcium / Q000378:metabolism*; D002316:Cardiotonic Agents / Q000633:toxicity; D002470:Cell Survival; D002633:Chemotaxis / Q000187:drug effects; D000242:Cyclic AMP / Q000378:metabolism; D054703:Cyclic Nucleotide Phosphodiesterases, Type 4; D016257:Fura-2 / Q000737:chemistry; D006801:Humans; D007093:Imidazoles / Q000633:toxicity; D009240:N-Formylmethionine Leucyl-Phenylalanine / Q000494:pharmacology; D009504:Neutrophils / Q000187:drug effects / Q000201:enzymology* / Q000378:metabolism; D010276:Parasympatholytics / Q000633:toxicity; D010726:Phosphodiesterase Inhibitors / Q000633:toxicity*; D010727:Phosphoric Diester Hydrolases / Q000187:drug effects* / Q000378:metabolism; D011719:Pyrazines / Q000633:toxicity; D011760:Pyrrolidinones / Q000633:toxicity; D020889:Rolipram; D013849:Thimerosal / Q000633:toxicity
D002316:Cardiotonic Agents; D007093:Imidazoles; D010276:Parasympatholytics; D010726:Phosphodiesterase Inhibitors; D011719:Pyrazines; D011760:Pyrrolidinones; C080660:6-bromo-8-methylaminoimidazo(1,2-a)pyrazine-2-carbonitrile; D013849:Thimerosal; D009240:N-Formylmethionine Leucyl-Phenylalanine; C048350:5-(4-acetamidophenyl)pyrazin-2(1H)-one; D000242:Cyclic AMP; D010727:Phosphoric Diester Hydrolases; D015105:3',5'-Cyclic-AMP Phosphodiesterases; D054703:Cyclic Nucleotide Phosphodiesterases, Type 4; D020889:Rolipram; D002118:Calcium; D016257:Fura-2