Defibrotide inhibits Ca2+ dependent neutrophil activation: implications for its pharmacological activity in vascular disorders.
Defibrotide (DEF) is a polydeoxyribonucleotide agent provided with profibrinolytic and antithrombotic properties. Moreover, an antiischemic, cardioprotective effect of the drug has recently been demonstrated in experimental animals. Increasing evidence exists of the important role played by neutrophils in the development of tissue damage during chronic and acute ischemia and in the early phases of reperfusion. In order to evaluate whether the overall cytoprotective effect of DEF could be based, at least in part, on a neutrophil-involving mechanism, the authors studied the in vitro effects of the drug on human neutrophil activation triggered by several specific stimuli. The drug dose-dependently (10-100 microM) inhibited enzyme release, superoxide anion generation, and chemiluminescence induced in neutrophils by the chemoattractant oligopeptide fMLP and by the divalent cation ionophores A23187 and ionomycin. The increase of extracellular calcium concentration from 0.5 to 2.0 mM antagonized the inhibitory effect of DEF. The use of the fluorescent probe Fura 2/AM led them to show that DEF is able to reduce the cytosolic free calcium increase following specific stimulation by affecting extracellular calcium entrance. Such a behavior resembles that of calcium-antagonistic drugs, thus suggesting that DEF works, at least in part, similarly to calcium entry blockers. Such an activity on cell calcium translocation could represent the underlying molecular mechanism of cytoprotection. Finally, the inhibitory action on neutrophil functions may play a role in tissue protection during ischemic injury.
Di Perri
T
T
Institute of Clinical Medicine, Università di Siena, Policinico Le Scotte, Italy.
Pasini
F L
FL
Ceccatelli
L
L
Pasqui
A L
AL
Capecchi
P L
PL
D002118:Calcium / Q000502:physiology*; D004305:Dose-Response Relationship, Drug; D005343:Fibrinolytic Agents / Q000494:pharmacology* / Q000627:therapeutic use; D005609:Free Radicals; D006801:Humans; D008163:Luminescent Measurements; D009113:Muramidase / Q000032:analysis; D009504:Neutrophils / Q000187:drug effects* / Q000201:enzymology / Q000502:physiology; D011089:Polydeoxyribonucleotides / Q000494:pharmacology* / Q000627:therapeutic use; D013481:Superoxides; D014652:Vascular Diseases / Q000188:drug therapy
D005343:Fibrinolytic Agents; D005609:Free Radicals; D011089:Polydeoxyribonucleotides; D013481:Superoxides; C036901:defibrotide; D009113:Muramidase; D002118:Calcium