Statin potentiates human platelet eNOS activity without enhancing eNOS mRNA and protein levels

Cerebrovasc Dis. 2008;26(2):190-8. doi: 10.1159/000145327. Epub 2008 Jul 15.

Abstract

Background/aims: Experimental studies suggest an enhanced endothelial and platelet nitric oxide (NO) generation after statin treatment, possibly due to increased endothelial NO synthase (eNOS) activity and protein levels. In parallel with experimental research, statins were shown to increase the forearm blood flow independently of serum cholesterol in humans. However, it was not possible to correlate blood flow changes with eNOS levels in these studies due to limitations in obtaining arterial samples. Hence, we investigated changes in eNOS activity, mRNA and protein levels after statin treatment in human platelets, which are readily accessible unlike arteries.

Methods: In vitro bleeding times were measured in 22 patients by stimulating platelets with collagen-epinephrine or collagen-ADP. To assess platelet eNOS activity, the bleeding times were also determined after incubating platelets with L-arginine. The measurements were repeated following 14 days of pravastatin (40 mg/day) treatment. Platelet-rich plasma was collected before and after statin treatment to evaluate eNOS mRNA (semiquantitative RT-PCR) and protein levels (Western blotting).

Results: The basal bleeding time was prolonged by 24 +/- 3% (mean +/- SE) when the samples were incubated with 500 microM of L-arginine. The NOS inhibitor L-N(5)-(I-iminoethyl)ornithine reversed this effect, suggesting that it was mediated by NO. After statin treatment, the NO-mediated prolongation of the bleeding time with 500 microM of L-arginine was significantly potentiated (to 44 +/- 10%). Despite enhanced eNOS activity, there was no significant change in platelet eNOS mRNA and protein levels after statin treatment.

Conclusion: These data demonstrate that platelet eNOS activity is potentiated after statin treatment in humans in parallel with experimental studies.

Publication types

  • Controlled Clinical Trial
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adult
  • Aged
  • Aged, 80 and over
  • Arginine / metabolism
  • Bleeding Time
  • Blood Platelets / drug effects*
  • Blood Platelets / enzymology
  • Brain Ischemia / blood
  • Brain Ischemia / complications*
  • Brain Ischemia / drug therapy
  • Enzyme Inhibitors / pharmacology
  • Female
  • Humans
  • Hydroxymethylglutaryl-CoA Reductase Inhibitors / therapeutic use*
  • Male
  • Middle Aged
  • Nitric Oxide / metabolism*
  • Nitric Oxide Synthase Type III / antagonists & inhibitors
  • Nitric Oxide Synthase Type III / genetics
  • Nitric Oxide Synthase Type III / metabolism*
  • Ornithine / analogs & derivatives
  • Ornithine / pharmacology
  • Platelet Aggregation / drug effects*
  • Pravastatin / therapeutic use*
  • RNA, Messenger / metabolism
  • Stroke / blood
  • Stroke / drug therapy*
  • Stroke / etiology
  • Treatment Outcome

Substances

  • Enzyme Inhibitors
  • Hydroxymethylglutaryl-CoA Reductase Inhibitors
  • RNA, Messenger
  • Nitric Oxide
  • N(G)-iminoethylornithine
  • Arginine
  • Ornithine
  • NOS3 protein, human
  • Nitric Oxide Synthase Type III
  • Pravastatin