Journal Mobile Options
Table of Contents
Vol. 26, No. 2, 2008
Issue release date: August 2008
Cerebrovasc Dis 2008;26:190–198
(DOI:10.1159/000145327)

Statin Potentiates Human Platelet eNOS Activity without Enhancing eNOS mRNA and Protein Levels

Yemisci M. · Ay H. · Kocaefe Ç. · Qui J. · Topalkara K. · Özgüç M. · Kirazli Ş. · Özcebe O. · Moskowitz M.A. · Dalkara T.
To view the fulltext, log in and/or choose pay-per-view option

Individual Users: Register with Karger Login Information

Please create your User ID & Password





Contact Information











I have read the Karger Terms and Conditions and agree.

To view the fulltext, please log in

To view the pdf, please log in

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 µM of L-arginine. The NOS inhibitor L-N5-(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 µM 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.



Copyright / Drug Dosage

Copyright: All rights reserved. No part of this publication may be translated into other languages, reproduced or utilized in any form or by any means, electronic or mechanical, including photocopying, recording, microcopying, or by any information storage and retrieval system, without permission in writing from the publisher or, in the case of photocopying, direct payment of a specified fee to the Copyright Clearance Center.
Drug Dosage: The authors and the publisher have exerted every effort to ensure that drug selection and dosage set forth in this text are in accord with current recommendations and practice at the time of publication. However, in view of ongoing research, changes in goverment regulations, and the constant flow of information relating to drug therapy and drug reactions, the reader is urged to check the package insert for each drug for any changes in indications and dosage and for added warnings and precautions. This is particularly important when the recommended agent is a new and/or infrequently employed drug.
Disclaimer: The statements, opinions and data contained in this publication are solely those of the individual authors and contributors and not of the publishers and the editor(s). The appearance of advertisements or/and product references in the publication is not a warranty, endorsement, or approval of the products or services advertised or of their effectiveness, quality or safety. The publisher and the editor(s) disclaim responsibility for any injury to persons or property resulting from any ideas, methods, instructions or products referred to in the content or advertisements.

References

  1. Amarenco P, Lavallee P, Touboul PJ: Statins and stroke prevention. Cerebrovasc Dis 2004;17(suppl 1):81–88.
  2. Plehn JF, Davis BR, Sacks FM, Rouleau JL, Pfeffer MA, Bernstein V, Cuddy TE, Moye LA, Piller LB, Rutherford J, Simpson LM, Braunwald E, The CARE Investigators: Reduction of stroke incidence after myocardial infarction with pravastatin: The Cholesterol and Recurrent Events (CARE) study. Circulation 1999;99:216–223.
  3. Randomised trial of cholesterol lowering in 4444 patients with coronary heart disease: The Scandinavian Simvastatin Survival Study (4S). Lancet 1994;344:1383–1389.
  4. Long-term effectiveness and safety of pravastatin in 9014 patients with coronary heart disease and average cholesterol concentrations: The LIPID trial follow-up. Lancet 2002;359:1379–1387.
  5. Vaughan CJ: Prevention of stroke and dementia with statins: effects beyond lipid lowering. Am J Cardiol 2003;91:23B–29B.
  6. Laufs U: Beyond lipid-lowering: effects of statins on endothelial nitric oxide. Eur J Clin Pharmacol 2003;58:719–731.
  7. Mason RP, Walter MF, Jacob RF: Effects of HMG-CoA reductase inhibitors on endothelial function: role of microdomains and oxidative stress. Circulation 2004;109:II34–II41.

    External Resources

  8. Mason JC: Statins and their role in vascular protection. Clin Sci (Lond) 2003;105:251–266.
  9. Laufs U, La Fata V, Plutzky J, Liao JK: Upregulation of endothelial nitric oxide synthase by HMG CoA reductase inhibitors. Circulation 1998;97:1129–1135.
  10. Endres M, Laufs U, Huang Z, Nakamura T, Huang P, Moskowitz MA, Liao JK: Stroke protection by 3-hydroxy-3-methylglutaryl (HMG)-CoA reductase inhibitors mediated by endothelial nitric oxide synthase. Proc Natl Acad Sci USA 1998;95:8880–8885.
  11. Laufs U, Gertz K, Huang P, Nickenig G, Bohm M, Dirnagl U, Endres M: Atorvastatin upregulates type III nitric oxide synthase in thrombocytes, decreases platelet activation, and protects from cerebral ischemia in normocholesterolemic mice. Stroke 2000;31:2442–2449.
  12. Yamada M, Huang Z, Dalkara T, Endres M, Laufs U, Waeber C, Huang PL, Liao JK, Moskowitz MA: Endothelial nitric oxide synthase-dependent cerebral blood flow augmentation by l-arginine after chronic statin treatment. J Cereb Blood Flow Metab 2000;20:709–717.
  13. Kaesemeyer WH, Caldwell RB, Huang J, Caldwell RW: Pravastatin sodium activates endothelial nitric oxide synthase independent of its cholesterol-lowering actions. J Am Coll Cardiol 1999;33:234–241.
  14. Amin-Hanjani S, Stagliano NE, Yamada M, Huang PL, Liao JK, Moskowitz MA: Mevastatin, an HMG-CoA reductase inhibitor, reduces stroke damage and upregulates endothelial nitric oxide synthase in mice. Stroke 2001;32:980–986.
  15. John S, Delles C, Jacobi J, Schlaich MP, Schneider M, Schmitz G, Schmieder RE: Rapid improvement of nitric oxide bioavailability after lipid-lowering therapy with cerivastatin within two weeks. J Am Coll Cardiol 2001;37:1351–1358.
  16. Laufs U, Wassmann S, Hilgers S, Ribaudo N, Bohm M, Nickenig G: Rapid effects on vascular function after initiation and withdrawal of atorvastatin in healthy, normocholesterolemic men. Am J Cardiol 2001;88:1306–1307.
  17. O’Driscoll G, Green D, Taylor RR: Simvastatin, an HMG-coenzyme A reductase inhibitor, improves endothelial function within 1 month. Circulation 1997;95:1126–1131.
  18. Masumoto A, Hirooka Y, Hironaga K, Eshima K, Setoguchi S, Egashira K, Takeshita A: Effect of pravastatin on endothelial function in patients with coronary artery disease (cholesterol-independent effect of pravastatin). Am J Cardiol 2001;88:1291–1294.
  19. Wassmann S, Ribaudo N, Faul A, Laufs U, Bohm M, Nickenig G: Effect of atorvastatin 80 mg on endothelial cell function (forearm blood flow) in patients with pretreatment serum low-density lipoprotein cholesterol levels <130 mg/dl. Am J Cardiol 2004;93:84–88.
  20. Dangas G, Badimon JJ, Smith DA, Unger AH, Levine D, Shao JH, Meraj P, Fier C, Fallon JT, Ambrose JA: Pravastatin therapy in hyperlipidemia: effects on thrombus formation and the systemic hemostatic profile. J Am Coll Cardiol 1999;33:1294–1304.
  21. Hale LP, Craver KT, Berrier AM, Sheffield MV, Case LD, Owen J: Combination of fosinopril and pravastatin decreases platelet response to thrombin receptor agonist in monkeys. Arterioscler Thromb Vasc Biol 1998;18:1643–1646.
  22. Kearney D, Fitzgerald D: The anti-thrombotic effects of statins. J Am Coll Cardiol 1999;33:1305–1307.
  23. Huhle G, Abletshauser C, Mayer N, Weidinger G, Harenberg J, Heene DL: Reduction of platelet activity markers in type II hypercholesterolemic patients by a HMG-CoA reductase inhibitor. Thromb Res 1999;95:229–234.
  24. Lacoste L, Lam JY, Hung J, Letchacovski G, Solymoss CB, Waters D: Hyperlipidemia and coronary disease: correction of the increased thrombogenic potential with cholesterol reduction. Circulation 1995;92:3172–3177.
  25. Tannous M, Cheung R, Vignini A, Mutus B: Atorvastatin increases ecNOS levels in human platelets of hyperlipidemic subjects. Thromb Haemost 1999;82:1390–1394.
  26. Adams HP Jr, Bendixen BH, Kappelle LJ, Biller J, Love BB, Gordon DL, Marsh EE 3rd: Classification of subtype of acute ischemic stroke: definitions for use in a multicenter clinical trial. TOAST. Trial of Org 10172 in Acute Stroke Treatment. Stroke 1993;24:35–41.
  27. Kratzer MA, Bellucci S, Caen JP: Detection of abnormal platelet functions with an in vitro model of primary haemostasis. Haemostasis 1985;15:363–370.
  28. Kundu SK, Heilmann EJ, Sio R, Garcia C, Davidson RM, Ostgaard RA: Description of an in vitro platelet function analyzer – PFA-100. Semin Thromb Hemost 1995;21:106–112.
  29. Anfossi G, Russo I, Massucco P, Mattiello L, Perna P, Tassone F, Trovati M: L-arginine modulates aggregation and intracellular cyclic 3,5-guanosine monophosphate levels in human platelets: direct effect and interplay with antioxidative thiol agent. Thromb Res 1999;94:307–316.
  30. Mammen EF, Comp PC, Gosselin R, Greenberg C, Hoots WK, Kessler CM, Larkin EC, Liles D, Nugent DJ: Pfa-100 system: a new method for assessment of platelet dysfunction. Semin Thromb Hemost 1998;24:195–202.
  31. Cannon CP, Braunwald E, McCabe CH, Rader DJ, Rouleau JL, Belder R, Joyal SV, Hill KA, Pfeffer MA, Skene AM: Intensive versus moderate lipid lowering with statins after acute coronary syndromes. N Engl J Med 2004;350:1495–1504.
  32. Liao JK: Role of statin pleiotropism in acute coronary syndromes and stroke. Int J Clin Pract Suppl 2003:51–57.

    External Resources

  33. Tschopp TB, Baumgartner HR, Silberbauer K, Sinzinger H: Platelet adhesion and platelet thrombus formation on subendothelium of human arteries and veins exposed to flowing blood in vitro: a comparison with rabbit aorta. Haemostasis 1979;8:19–29.
  34. Radomski MW, Palmer RM, Moncada S: Characterization of the L-arginine: nitric oxide pathway in human platelets. Br J Pharmacol 1990;101:325–328.
  35. Radomski MW, Palmer RM, Moncada S: Modulation of platelet aggregation by an L-arginine-nitric oxide pathway. Trends Pharmacol Sci 1991;12:87–88.
  36. Bode-Boger SM, Boger RH, Creutzig A, Tsikas D, Gutzki FM, Alexander K, Frolich JC: L-arginine infusion decreases peripheral arterial resistance and inhibits platelet aggregation in healthy subjects. Clin Sci (Lond) 1994;87:303–310.
  37. Weber AA, Strobach H, Schror K: Direct inhibition of platelet function by organic nitrates via nitric oxide formation. Eur J Pharmacol 1993;247:29–37.
  38. Marietta M, Castelli I, Piccinini F, Neri I, Bertesi M, Facchinetti F, Torelli G: The PFA-100 system for the assessment of platelet function in normotensive and hypertensive pregnancies. Clin Lab Haematol 2001;23:131–134.
  39. Muller I, Besta F, Schulz C, Li Z, Massberg S, Gawaz M: Effects of statins on platelet inhibition by a high loading dose of clopidogrel. Circulation 2003;108:2195–2197.
  40. Dajani EZ, Shahwan TG, Dajani NE: Statins, platelet aggregation and coronary heart disease. J Assoc Acad Minor Phys 2002;13:27–31.

    External Resources

  41. Hernandez-Perera O, Perez-Sala D, Navarro-Antolin J, Sanchez-Pascuala R, Hernandez G, Diaz C, Lamas S: Effects of the 3- hydroxy-3-methylglutaryl-CoA reductase inhibitors, atorvastatin and simvastatin, on the expression of endothelin-1 and endothelial nitric oxide synthase in vascular endothelial cells. J Clin Invest 1998;101:2711–2719.
  42. Mital S, Zhang X, Zhao G, Bernstein RD, Smith CJ, Fulton DL, Sessa WC, Liao JK, Hintze TH: Simvastatin upregulates coronary vascular endothelial nitric oxide production in conscious dogs. Am J Physiol Heart Circ Physiol 2000;279:H2649–H2657.
  43. McGirt MJ, Lynch JR, Parra A, Sheng H, Pearlstein RD, Laskowitz DT, Pelligrino DA, Warner DS: Simvastatin increases endothelial nitric oxide synthase and ameliorates cerebral vasospasm resulting from subarachnoid hemorrhage. Stroke 2002;33:2950– 2956.
  44. Tseng MY, Czosnyka M, Richards H, Pickard JD, Kirkpatrick PJ: Effects of acute treatment with pravastatin on cerebral vasospasm, autoregulation, and delayed ischemic deficits after aneurysmal subarachnoid hemorrhage: a phase II randomized placebo-controlled trial. Stroke 2005;36:1627–1632.
  45. Lynch JR, Wang H, McGirt MJ, Floyd J, Friedman AH, Coon AL, Blessing R, Alexander MJ, Graffagnino C, Warner DS, Laskowitz DT: Simvastatin reduces vasospasm after aneurysmal subarachnoid hemorrhage: results of a pilot randomized clinical trial. Stroke 2005;36:2024–2026.
  46. Garg UC, Hassid A: Nitric oxide-generating vasodilators and 8-bromo-cyclic guanosine monophosphate inhibit mitogenesis and proliferation of cultured rat vascular smooth muscle cells. J Clin Invest 1989;83:1774–1777.
  47. Peng HB, Libby P, Liao JK: Induction and stabilization of I kappa B alpha by nitric oxide mediates inhibition of NF-kappa B. J Biol Chem 1995;270:14214–14219.
  48. Kubes P, Suzuki M, Granger DN: Nitric oxide: an endogenous modulator of leukocyte adhesion. Proc Natl Acad Sci USA 1991;88:4651–4655.
  49. Lefer AM, Ma XL: Decreased basal nitric oxide release in hypercholesterolemia increases neutrophil adherence to rabbit coronary artery endothelium. Arterioscler Thromb 1993;13:771–776.
  50. Radomski MW, Palmer RM, Moncada S: Endogenous nitric oxide inhibits human platelet adhesion to vascular endothelium. Lancet 1987;2:1057–1058.
  51. Jayachandran M, Mukherjee R, Steinkamp T, Labreche P, Bracamonte MP, Okano H, Owen WG, Miller VM: Differential effects of 17beta-estradiol, conjugated equine estrogen and raloxifene on mRNA, aggregation and secretion in platelets. Am J Physiol Heart Circ Physiol 2005;288:H2355–H2362.
  52. Baker GR, Sullam PM, Levin J: A simple, fluorescent method to internally label platelets suitable for physiological measurements. Am J Hematol 1997;56:17–25.
  53. Laufs U, Gertz K, Dirnagl U, Bohm M, Nickenig G, Endres M: Rosuvastatin, a new HMG-CoA reductase inhibitor, upregulates endothelial nitric oxide synthase and protects from ischemic stroke in mice. Brain Res 2002;942:23–30.
  54. Sessa WC: eNOS at a glance. J Cell Sci 2004;117:2427–2429.
  55. LaRosa JC: Pleiotropic effects of statins and their clinical significance. Am J Cardiol 2001;88:291–293.
  56. Libby P, Aikawa M: Stabilization of atherosclerotic plaques: new mechanisms and clinical targets. Nat Med 2002;8:1257–1262.
  57. Vaughan CJ, Gotto AM Jr, Basson CT: The evolving role of statins in the management of atherosclerosis. J Am Coll Cardiol 2000;35:1–10.
  58. Ledoux S, Laouari D, Essig M, Runembert I, Trugnan G, Michel JB, Friedlander G: Lovastatin enhances ecto-5′-nucleotidase activity and cell surface expression in endothelial cells: implication of Rho-family GTPases. Circ Res 2002;90:420–427.


Pay-per-View Options
Direct payment This item at the regular price: USD 38.00
Payment from account With a Karger Pay-per-View account (down payment USD 150) you profit from a special rate for this and other single items.
This item at the discounted price: USD 26.50