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Thromb Haemost 1997; 77(05): 1020-1024
DOI: 10.1055/s-0038-1656096
Animal models
Schattauer GmbH Stuttgart

Erythropoietin Potentiates Thrombus Development in a Canine Arterio-Venous Shunt Model

Roman F Wolf
The Department of Medicine and W. K. Warren Medical Research Institute, University of Oklahoma Health Sciences Center, Oklahoma City, Oklahoma, USA
,
Laura S Gilmore
The Department of Medicine and W. K. Warren Medical Research Institute, University of Oklahoma Health Sciences Center, Oklahoma City, Oklahoma, USA
,
Paul Friese
The Department of Medicine and W. K. Warren Medical Research Institute, University of Oklahoma Health Sciences Center, Oklahoma City, Oklahoma, USA
,
Tamyra Downs
The Department of Medicine and W. K. Warren Medical Research Institute, University of Oklahoma Health Sciences Center, Oklahoma City, Oklahoma, USA
,
Samuel A Burstein
The Department of Medicine and W. K. Warren Medical Research Institute, University of Oklahoma Health Sciences Center, Oklahoma City, Oklahoma, USA
,
George L Dale
The Department of Medicine and W. K. Warren Medical Research Institute, University of Oklahoma Health Sciences Center, Oklahoma City, Oklahoma, USA
› Author Affiliations
Further Information

Publication History

Received 16 September 1996

Accepted after resubmission 06 January 1997

Publication Date:
26 July 2018 (online)

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Summary

Erythropoietin (EPO) has been previously shown to affect platelet as well as red cell production. In addition, recent studies demonstrated that platelets from EPO-treated dogs are hyperreactive towards thrombin when compared to age-matched, control platelets. This report extends these observations by quantitating the thrombogenic potential of EPO in dogs. Dogs with arterio-venous (A-V) shunts received 100 U EPO/kg/day for 6 days, and thrombogenicity was serially monitored by insertion of a thrombotic surface into the A-V shunt. The resulting experimental thrombi were analyzed for platelet and erythrocyte content after formalin-fixation and chymotrypsin digestion, a technique which allows non-isotopic quantitation of cellular components. By day 5 of EPO-administration all animals demonstrated a significant increase in platelet and red cell content of the experimental thrombi; the average increase in platelet number was 2.94 ±0.12 fold (mean ± 1 SE; n = 3; p = 0.006) above baseline while that for red cells was 2.46 ±0.18 fold above baseline (p = 0.023). After cessation of EPO, thrombogenicity returned to normal. During EPO-treatment, the percentage of thiazole orange-positive (TO+) platelets increased significantly to 17.2 ± 1.6% (mean ± 1 SE; n = 3) on day 5 compared to a pre-treatment level of 8.5 ± 0.9% (p = 0.029). Although the percentage of TO+ erythrocytes also increased during the short course of EPO administration, the change was not significant. Despite the increases in TO+ cells, total platelet and erythrocyte counts did not change significantly within the time frame of these experiments. Fibrin/fibrinogen content of the experimental thrombi was unaltered with EPO-treatment. These data demonstrate that human EPO is pro-thrombotic in dogs and, in conjunction with earlier studies, suggest that hyperreactive platelets may be responsible for the potentiated thrombogenicity.

 

  • References

  • 1 Krantz SB. Erythropoietin. Blood 1991; 77: 419-434
    PubMedSearch in Google Scholar
  • 2 Eschbach JW, Abudulhadi MH, Browne JK, Delano BG, Downing MR, Egrie JC, Evans RW, Friedman EA, Graber SE, Haley NR, Korbert S, Krantz SB, Lundin AP, Nissenson AR, Ogden DA, Paganini EP, Rader B, Rutsky EA, Stivelman J, Stone WJ, Teschan P, VanStone JC, VanWyck DB, Zuckerman K. Adamson JW. Recombinant human erythropoietin in anemic patients with end-stage renal disease. Annals Int Med 1989; 111: 992-1000
    CrossrefPubMedSearch in Google Scholar
  • 3 Erslev AJ, Wilson J, Caro J. Erythropoietin titers in anemic, nonuremic patients. J Lab Clin Med 1987; 109: 429-433
    PubMedSearch in Google Scholar
  • 4 Youssoufian H, Longmore G, Neumann D, Yoshimura A, Lodish HF. Structure, function and activation of the erythropoietin receptor. Blood 1993; 81: 2223-2236
    PubMedSearch in Google Scholar
  • 5 Lacombe C, Dusanter I, Gobert S, Muller O, Gisselbrecht S, Mayeux P. Intracellular pathways activated by erythropoietin. Annals N Y Acad Sci 1994; 718: 223-230
    PubMedSearch in Google Scholar
  • 6 Fraser JK, Tan AS, Lin FK, Berridge MV. Expression of specific high- affinity binding sites for erythropoietin on rat and mouse megakaryocytes. ExpHematol 1989; 17: 10-16
    PubMedSearch in Google Scholar
  • 7 Ishibashi T, Koziol JA. Burstein SA. Human recombinant erythropoietin promotes differentiation of murine megakaryocytes in vitro. J Clin Invest 1987; 79: 286-289
    CrossrefPubMedSearch in Google Scholar
  • 8 Berridge MV, Fraser JK, Carter JM, Lin FK. Effects of recombinant human erythropoietin on megakaryoctyes and on platelet production in the rat. Blood 1988; 72: 970-977
    PubMedSearch in Google Scholar
  • 9 Shikama Y, Ishibashi T, Kimura H, Kawaguchi M, Uchida T, Maruyama Y. Transient effect of erythropoietin on thrombocytopoiesis in vivo in mice. Exp Hematol 1992; 20: 216-222
    PubMedSearch in Google Scholar
  • 10 Tsukada J, Misago M, Kikuchi M, Sato T, Ogawa R, Oda S, Chiba S, Eto S. The effect of high doses of recombinant human erythropoietin on mega-karyocytopoiesis and platelet production in splenectomized mice. Br J Haematol 1990; 76: 260-268
    CrossrefPubMedSearch in Google Scholar
  • 11 Kitagawa S, Saito M, Miura Y. Recombinant human erythropoietin at high doses stimulates thrombopoiesis: Treatment for protracted severe myelo-suppression complicating interferon-α and busulfan therapy for chronic myelogenous leukaemia. Eur J Hematol 1995; 55: 285-286
    PubMedSearch in Google Scholar
  • 12 Pirisi M, Fabris C, Soardo G, Cecchin E, Toniutto P, Bartoli E. Thrombocytopenia of chronic liver disease corrected by erythropoietin treatment. J Hepatology 1994; 21: 376-380
    CrossrefPubMedSearch in Google Scholar
  • 13 Sharpe PC, Desai ZR, Morris TCM. Increase in mean platelet volume in patients with chronic renal failure treated with erythropoietin. J Clin Pathol 1994; 47: 159-161
    CrossrefPubMedSearch in Google Scholar
  • 14 vanGeet C, Hauglustaine D, Verresen L, Vanrusselt M, Vermylen J. Haemostatic effects of recombinant human erythropoietin in chronic haemodialysis patients. Thromb Haemost 1989; 61: 117-121
    Thieme ConnectPubMedSearch in Google Scholar
  • 15 Cases A, Escolar G, Reverter JC, Ordinas A, Lopez-Pedret J, Revert L, Castillo R. Recombinant human erythropoietin treatment improves platelet function in uremic patients. Kidney Intemat 1992; 42: 668-672
    PubMedSearch in Google Scholar
  • 16 Malyszko J, Malyszko JS, Borawski J, Rydzewski A, Kalinowski M, Azzadin A, Mysliwiec M, Buczko W. A study of platelet functions, some hemostatic and fibrinolytic parameters in relation to serotonin in hemo- dialyzed patients under erythropoietin therapy. Thrombosis Res 1995; 77: 133-143
    CrossrefPubMedSearch in Google Scholar
  • 17 Wolf RF, Peng J, Friese P, Gilmore LS, Burstein SA, Dale GL. Erythropoietin administration increases production and reactivity of platelets in dogs. Blood 1996; 88: 141b (Abstract).
    PubMedSearch in Google Scholar
  • 18 Zingg W, Ip WF, Sefton MV, Mancer K. A chronic arteriovenous shunt for the testing of biomaterials and devices in dogs. Life Support Systems 1986; 4: 221-229
    PubMedSearch in Google Scholar
  • 19 Sils IV, Szlyk-Modrow PC, Tartarini KA, Hubbard LJ, Glass E, Caretti DM, Darrigrand AA. Chronic implantation of nonocclusive catheters and flow probes in the splanchnic and hindlimb vasculature of the rabbit. Lab Anim Sci 1994; 44: 319-325
    PubMedSearch in Google Scholar
  • 20 Peters RF, Lees CM, Mitchell KA, Tweed MF, Talbot MD, Wallis RB. The characterization of thrombus development in an improved model of arterio-venous shunt thrombosis in the rat and the effects of recombinant desulphatohirudin (CPG 39393), heparin and iloprost. Thromb Haemost 1991; 65: 268-274
    Thieme ConnectPubMedSearch in Google Scholar
  • 21 Heilmann E, Hynes LA, Friese P, George JN, Burstein SA, Dale GL. Dog platelets accumulate intracellular fibrinogen as they age. J Cell Physiol 1994; 161: 23-30
    CrossrefPubMedSearch in Google Scholar
  • 22 Dale GL, Hynes LA, Wolf RF, Williams LS, Burstein SA. Non-isotopic method for quantitation of platelets and erythrocytes in experimental thrombi. Thromb Haemost 1996; 75: 668-673
    PubMedSearch in Google Scholar
  • 23 Dale GL, Friese P, Hynes LA, Burstein SA. Demonstration that thiazole orange-positive platelets in the dog are less than twenty-four hours old. Blood 1995; 85: 1822-1825
    PubMedSearch in Google Scholar
  • 24 Browne PV, Hebbel RP. CD36-positive stress reticulocytosis in sickle cell anemia. J Lab Clin Med 1996; 127: 340-347
    CrossrefPubMedSearch in Google Scholar
  • 25 Macdougall IC, Davies ME, Hallett I, Cochlin DL, Hutton RD, Coles GA, Williams JD. Coagulation studies and fistula blood flow during erythropoietin therapy in haemodialysis patients. Nephrology, Dialysis, Transplantation 1991; 6: 862-867
    CrossrefPubMedSearch in Google Scholar
  • 26 Kampf D, Kahl A, Passlick J, Pustelnik A, Eckardt KU, Ehmer B, Jacobs C, Baumelou A, Grabensee B, Gahl GM. Single-dose kinetics of recombinant human erythropoietin after intravenous, subcutaneous and intraperitoneal administration. Contrib Nephrol 1989; 76: 106-111
    PubMedSearch in Google Scholar
  • 27 Macdougall IC, Roberts DE, Neubert P, Dharmasena AD, Coles GA, Williams JD. Pharmacokinetics of intravenous, intraperitoneal and subcutaneous recombinant erythropoietin in patients with CAPD. Contrib Nephrol 1989; 76: 112-121
    PubMedSearch in Google Scholar
  • 28 Sherwood JB, Goldwasser E, Chilcote R, Carmichael LD, Nagel RL. Sickle cell anemia patients have low erythropoietin levels for their degree of anemia. Blood 1986; 67: 46-49
    PubMedSearch in Google Scholar
  • 29 Balkaran B, Char G, Morris JS, Thomas PW, Serjeant BE, Serjeant GR. Stroke in a cohort of patients with homozygous sickle cell disease. J Pediatrics 1992; 120: 360-366
    CrossrefPubMedSearch in Google Scholar
  • 30 Francis RB, Johnson CS. Vascular occlusion in sickle cell disease: Current concepts and unanswered questions. Blood 1991; 77: 1405-1414
    PubMedSearch in Google Scholar
  • 31 Mueller-Soyano A, De Roura ET, Duke PR, De AcquatellaGC, Arends T, Guinto E, Beutler E. Pyruvate kinase deficiency and leg ulcers. Blood 1976; 47: 807-813
    PubMedSearch in Google Scholar
  • 32 Palek J, Jarolim P, Hereditary spherocytosis. elliptocytosis and related disorders. In: Williams Hematology. Beutler E, Lichtman MA, Coller BS, Kipps TJ. eds. New York: McGraw-Hill; 1995. Ed. 5thpp 536-557
    Search in Google Scholar
  • 33 Macdougall IC, Hutton RD, Coles GA, Williams JD. The use of erythropoietin in renal failure. Postgrad Med J 1991; 67: 09-15
    CrossrefPubMedSearch in Google Scholar
  • 34 Winearls CG, Oliver DO, Pippard MJ, Reid C, Downing MR, Cotes PM. Effect of human erythropoietin derived from recombinant DNA on the anaemia of patients maintained by chronic haemodialysis. Lancet 1986; 8517: 1175-1177
    PubMedSearch in Google Scholar
  • 35 Canadian Erythropoietin Study Group. Association between recombinant human erythropoietin and quality of life and exercise capacity of patients receiving haemodialysis. BMJ 1990; 300: 573-8
    CrossrefPubMedSearch in Google Scholar