title: High-dose heparin decreases nitric oxide production by cultured bovine endothelial cells.
abstract: BACKGROUND
  Abrupt cessation of heparin therapy can lead to a recrudescence of thrombosis and acute ischemia. Endothelial NO is an important endogenous inhibitor of platelet-mediated thrombosis, yet biochemical studies examining the effect of heparin on NO production by the endothelium have heretofore been lacking.
  
  
  RESULTS
  In an attempt to address the effect of heparin on endothelial cell production of NO, confluent bovine aortic endothelial cells (BAECs) on microcarrier beads were incubated in the presence or absence of heparin. Results indicate that BAECs incubated with heparin were less able to inhibit platelet aggregation than control cells (P<.005 by ANOVA) and that this effect correlated with a decrease in NO production (36% decrease for heparin compared with control, P<.05). Dextran sulfate evoked the same response (67% decrease, P<.0001 compared with control), suggesting that the decrease in NO after heparin treatment is secondary to its negative charge rather than to a specific polysaccharide sequence. The decrease in NO production by heparin was accompanied by a 72% decrease in steady-state Nos 3 mRNA as well as a 49% decrease in immunodetectable endothelial NO synthase (eNOS) protein.
  
  
  CONCLUSIONS
  These data show that high-dose heparin at concentrations achieved in some acute cardiovascular settings increases in vitro platelet aggregation in media conditioned by endothelial cells by decreasing endothelial NO production through a mechanism that involves a decrease in steady-state Nos 3 mRNA and eNOS protein. These observations suggest a possible mechanism by which to explain in part the prothrombotic effects of heparin.
authors.affiliation: Whitaker Cardiovascular Institute and Evans Department of Medicine, Boston University School of Medicine, MA 02118, USA.
authors.forename: G R
authors.initials: GR
authors.lastname: Upchurch
authors.forename: G N
authors.initials: GN
authors.lastname: Welch
authors.forename: J E
authors.initials: JE
authors.lastname: Freedman
authors.forename: A J
authors.initials: AJ
authors.lastname: Fabian
authors.forename: A
authors.initials: A
authors.lastname: Pigazzi
authors.forename: A M
authors.initials: AM
authors.lastname: Scribner
authors.forename: C S
authors.initials: CS
authors.lastname: Alpert
authors.forename: J F
authors.initials: JF
authors.lastname: Keaney
authors.forename: J
authors.initials: J
authors.lastname: Loscalzo
mesh_terms: D000818:Animals; D000925:Anticoagulants / Q000494:pharmacology*; D001011:Aorta / Q000166:cytology; D001792:Blood Platelets / Q000378:metabolism; D002417:Cattle; D002478:Cells, Cultured; D006152:Cyclic GMP / Q000096:biosynthesis; D003864:Depression, Chemical; D016264:Dextran Sulfate / Q000494:pharmacology; D004730:Endothelium, Vascular / Q000187:drug effects* / Q000378:metabolism; D004790:Enzyme Induction / Q000187:drug effects; D006493:Heparin / Q000737:chemistry / Q000494:pharmacology*; D008863:Microspheres; D009569:Nitric Oxide / Q000096:biosynthesis*; D019001:Nitric Oxide Synthase / Q000096:biosynthesis / Q000235:genetics; D010974:Platelet Aggregation; D011479:Protamines / Q000494:pharmacology; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism
chemical_list: D000925:Anticoagulants; D011479:Protamines; D012333:RNA, Messenger; D009569:Nitric Oxide; D006493:Heparin; D016264:Dextran Sulfate; D019001:Nitric Oxide Synthase; D006152:Cyclic GMP