title: The influence of platelet-smooth muscle cell interaction on the oxidative modification of low-density lipoprotein.
abstract: OBJECTIVE
  In view of the recognized association between thrombosis and atherosclerosis, it is hypothesized that exposure of arterial smooth muscle cells (SMC) to thrombogenic agents such as platelets and thrombin will alter the oxidation of low-density lipoprotein (LDL) and that this effect may be diminished by thrombin inhibition.
  
  
  METHODS
  Quiescent human aortic SMC in culture were exposed to LDL (40 microg protein/ml) alone or with washed human platelets (5 x 10(6)/ml), thrombin (40 units/ml), or a combination of these agents for 48 h. The media were removed, and both media and cell lysate fractions were assayed for malondialdehyde (MDA) content as an index of oxidation. Isolated platelets exposed to LDL and thrombin were studied in a similar manner to determine their individual oxidative activity. Finally, SMC and platelets were incubated with LDL and varying concentrations of thrombin (10-80 units/ml), both alone and in the presence of the thrombin inhibitors hirudin (u/u), and heparin (u/u), and MDA was measured.
  
  
  RESULTS
  SMC and platelets each demonstrated an ability to oxidize LDL, increasing MDA concentrations by 1.8- (P < 0.05) and 4- (P < 0.01) fold, respectively, compared to lipid-free media. Both platelets (P < 0.05) and thrombin (P < 0.001) enhanced the oxidation of LDL by SMC, while a combination of these two agents resulted in an additive effect (P < 0.001). The SMC lysate fraction showed an increase in oxidative products following exposure to platelets (P < 0.01) but not thrombin, suggesting that platelets stimulated uptake of the oxidized lipid by the SMC. Isolated platelets responded to thrombin with an increase in MDA within the media (P < 0.001). Smooth muscle cells exposed to thrombin also showed a dose-dependent increase in LDL oxidation (P < 0.01). This effect was not altered by hirudin, but was significantly inhibited by heparin (P < 0.05).
  
  
  CONCLUSIONS
  These results indicate that the oxidative potential of SMC and platelets is enhanced by their coincubation and by their concurrent exposure to thrombin. Heparin appears to block thrombin-stimulated oxidation. This interaction could be relevant to the dynamic interaction between atherosclerosis and thrombogenesis.
authors.affiliation: Department of Surgery, Case Western Reserve University, Cleveland, Ohio 44109, USA.
authors.forename: J Jeffrey
authors.initials: JJ
authors.lastname: Alexander
authors.forename: Isabel
authors.initials: I
authors.lastname: Lewis
mesh_terms: D001011:Aorta / Q000166:cytology; D001161:Arteriosclerosis / Q000378:metabolism; D001792:Blood Platelets / Q000166:cytology* / Q000378:metabolism*; D002450:Cell Communication / Q000502:physiology; D002478:Cells, Cultured; D005343:Fibrinolytic Agents / Q000494:pharmacology; D006490:Hemostatics / Q000494:pharmacology; D006629:Hirudins / Q000494:pharmacology; D006801:Humans; D008077:Lipoproteins, LDL / Q000378:metabolism*; D008315:Malondialdehyde / Q000378:metabolism; D009131:Muscle, Smooth, Vascular / Q000166:cytology* / Q000378:metabolism*; D010084:Oxidation-Reduction; D013917:Thrombin / Q000494:pharmacology; D013927:Thrombosis / Q000378:metabolism
chemical_list: D005343:Fibrinolytic Agents; D006490:Hemostatics; D006629:Hirudins; D008077:Lipoproteins, LDL; C099252:oxidized low density lipoprotein; D008315:Malondialdehyde; D013917:Thrombin