title: Iron-dependent human platelet activation and hydroxyl radical formation: involvement of protein kinase C.
abstract: BACKGROUND
  Iron is an important modulator of lipid peroxidation, and its levels have been associated with the progression of atherosclerosis. Little is known about the possibility that this metal, when released from tissue stores, may modulate the reactivity of blood cell components, in particular platelets. Therefore, we investigated a possible link between iron, oxygen free radical formation, and platelet function.
  
  
  RESULTS
  Human whole blood was stimulated with collagen 2 micrograms/mL, and an irreversible aggregation with thromboxane (Tx)B2 formation was observed (15+/-4 versus 130+/-10 ng/mL). Deferoxamine (DSF), a specific iron chelator, and catalase, an H2O2 scavenger, inhibited collagen-induced whole-blood aggregation. The aggregation was accompanied by an increase in hydroxyl radical (OH.) levels (30+/-8 versus 205+/-20 nmol/L dihydroxybenzoates), which were reduced by DSF and by 2 specific OH. scavengers, mannitol and deoxyribose. Iron (Fe2+) dose-dependently induced platelet aggregation, TxB2 formation (6+/-2 versus 135+/-8 ng/mL), and protein kinase C (PKC) translocation from the cytosol to the cell membrane when added to platelets that have been primed with a low concentration of collagen (0.2 micrograms/mL). In the same system, an increase in OH. levels was observed (37+/-12 versus 230+/-20 nmol/L dihydroxybenzoates). Mannitol and deoxyribose, but not urea, were able to reduce OH. formation, PKC activation, and platelet aggregation. Selective inhibition of PKC activity by GF 109203X prevented iron-dependent platelet aggregation without influencing OH. production.
  
  
  CONCLUSIONS
  The present study shows that iron can directly interact with human platelets, resulting in their activation. Its action is mediated by OH. formation and involves PKC activity. Our findings provide an additional contribution to the understanding of the mechanism(s) by which iron overload might promote atherosclerosis and coronary artery disease.
authors.affiliation: Institute of Clinical Medicine I, University La Sapienza, the National Institute of Nutrition, Rome, Italy.
authors.forename: D
authors.initials: D
authors.lastname: Praticó
authors.forename: M
authors.initials: M
authors.lastname: Pasin
authors.forename: O P
authors.initials: OP
authors.lastname: Barry
authors.forename: A
authors.initials: A
authors.lastname: Ghiselli
authors.forename: G
authors.initials: G
authors.lastname: Sabatino
authors.forename: L
authors.initials: L
authors.lastname: Iuliano
authors.forename: G A
authors.initials: GA
authors.lastname: FitzGerald
authors.forename: F
authors.initials: F
authors.lastname: Violi
mesh_terms: D000328:Adult; D001241:Aspirin / Q000494:pharmacology; D001792:Blood Platelets / Q000187:drug effects / Q000201:enzymology; D002374:Catalase / Q000494:pharmacology; D002614:Chelating Agents / Q000494:pharmacology; D003094:Collagen / Q000494:pharmacology; D003676:Deferoxamine / Q000494:pharmacology; D003855:Deoxyribose / Q000494:pharmacology; D004234:Diuretics, Osmotic / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D004492:Edetic Acid / Q000494:pharmacology; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005260:Female; D006801:Humans; D017665:Hydroxyl Radical / Q000378:metabolism*; D007211:Indoles / Q000494:pharmacology; D007501:Iron / Q000378:metabolism / Q000494:pharmacology*; D007546:Isoquinolines / Q000494:pharmacology; D008297:Male; D008301:Maleimides / Q000494:pharmacology; D008353:Mannitol / Q000494:pharmacology; D015539:Platelet Activation / Q000187:drug effects* / Q000502:physiology*; D010974:Platelet Aggregation / Q000187:drug effects / Q000502:physiology; D010975:Platelet Aggregation Inhibitors / Q000494:pharmacology; D011493:Protein Kinase C / Q000037:antagonists & inhibitors / Q000378:metabolism*; D013449:Sulfonamides; D013928:Thromboxane A2 / Q000378:metabolism; D013929:Thromboxane B2 / Q000378:metabolism
chemical_list: D002614:Chelating Agents; D004234:Diuretics, Osmotic; D004791:Enzyme Inhibitors; D007211:Indoles; D007546:Isoquinolines; D008301:Maleimides; D010975:Platelet Aggregation Inhibitors; D013449:Sulfonamides; D017665:Hydroxyl Radical; D008353:Mannitol; D003855:Deoxyribose; D013929:Thromboxane B2; D013928:Thromboxane A2; D003094:Collagen; D004492:Edetic Acid; D007501:Iron; D002374:Catalase; D011493:Protein Kinase C; D003676:Deferoxamine; C070515:bisindolylmaleimide I; C063509:N-(2-(4-bromocinnamylamino)ethyl)-5-isoquinolinesulfonamide; D001241:Aspirin