Therapeutic Effect of a Low Molecular Weight Dermatan Sulphate (Desmin 370) in Rat Venous Thrombosis - Evidence for an Anticoagulant-Independent Mechanism

 

PDF Download Buy Article Permissions and Reprints

Summary

We evaluated the capacity of a low molecular weight dermatan sulphate (D370) to prevent thrombus formation and to induce a reduction of a stabilized thrombus in a rat venous thrombosis model. Injection of D370, 10 min before induction of venous stasis (prevention model), prevented thrombus formation in a dose-dependent way (ED₅₀: 2.3 mg/kg). When given to rats 6 h after induction of venous stasis (therapeutic model), D370 caused a time- and dose-dependent reduction in thrombus size (60% to 70% reduction 2 h after injection of 10 mg/kg). At comparable antithrombotic dosages (i.e. minimum dose giving complete inhibition of thrombus formation), heparin (0.5 mg/kg) only caused 40% reduction of a preformed thrombus while hirudin (1 mg/kg) was virtually ineffective (less than 10% reduction in weight). All three compounds inhibited ¹²⁵I-fibrin(ogen) deposition on 6-h aged thrombi by more than 85%, suggesting that D370 and, to a lesser extent, heparin reduce thrombus size via mechanisms other than inhibition of thrombus accretion. The involvement of a fibrinolysis-mediated mechanism in the D370-induced effect is suggested by the following. EACA (1 g/kg), when given to thrombus-bearing control animals, did not influence thrombus weight. However, when administered before D370 treatment, it prevented the expected reduction in thrombus weight by more than 80%, without influencing the effect of D370 on ¹²⁵I-fibrin(ogen) accumulation onto preexisting thrombi. D370 injection caused neither an enhancement of fibrinolytic activity nor a reduction of PAI in plasma. In vitro, D370 (200 μg/ml) was unable to potentiate the spontaneous or PA-induced lysis of ¹²⁵I-fibrinogen labelled blood, plasma, or purified fibrin clots. It is suggested that prevention of thrombus formation by D370 is related mainly to inhibition of blood coagulation, whereas reduction of the weight of aged thrombi is primarily due to an anticoagulant-independent mechanism, most probably involving local enhancement of the fibrinolytic process. D370 may represent an alternative pharmacologic agent both in the prevention and in the therapy of venous thrombosis.

• References

• 1 Tollefsen DM, Pestka CA, Monafo WJ. Activation of heparin cofactor II by dermatan sulfate. J Biol Chem 1983; 258: 6713-6716
  PubMedGoogle Scholar
• 2 Ofosu FA, Modi GJ, Smith LM, Cerksus AL, Hirsh J, Blajchman MA. Heparan sulfate and dermatan sulfate inhibit the generation of thrombin activity in plasma by complementary pathways. Blood 1984; 54: 742-749
  PubMedGoogle Scholar
• 3 Ofosu FA, Blajchman MA, Modi GJ, Smith LM, Buchanan MR, Hirsh J. The importance of thrombin inhibition for the expression of the anticoagulant activities of heparin, dermatan sulphate, low molecular weight heparin and pentosan polysulphate. Br J Haematol 1985; 60: 695-704
  CrossrefPubMedGoogle Scholar
• 4 Fernandez F, Van Ryn J, Ofosu FA, Hirsh J, Buchanan MR. The haemorrhagic and antithrombotic effects of dermatan sulphate. Br J Haematol 1986; 64: 309-317
  CrossrefPubMedGoogle Scholar
• 5 Merton RE, Thomas DP. Experimental studies on the relative efficacy of dermatan sulphate and heparin as antithrombotic agents. Thromb Haemostas 1987; 58: 839-842
  PubMedGoogle Scholar
• 6 Maggi A, Abbadini M, Pagella PG, Borowska A, Pangrazzi J, Donati MB. Antithrombotic properties of dermatan sulphate in a rat venous thrombosis model. Haemostasis 1987; 17: 329-335
  PubMedGoogle Scholar
• 7 Van Ryn-McKenna J, Ofosu FA, Gray E, Hirsh J, Buchanan MR. The effects of dermatan sulfate and heparin on inhibition of thrombus growth in vivo. Ann NY Acad Sci USA 1989; 556: 304-312
  CrossrefPubMedGoogle Scholar
• 8 Fernandez F, Buchanan MR, Hirsh J, Fenton JW, Ofosu F. Catalysis of thrombin inhibition provides an index for estimating the antithrombotic potential of glycosaminoglycans in rabbit. Thromb Haemostas 1987; 57: 286-293
  PubMedGoogle Scholar
• 9 Ofosu FA, Fernandez F, Anvari N, Caranobe C, Dol F, Cadroy Y, Petitou M, Mardiguian J, Sie P, Boneu B. Further studies on the mechanisms for the antithrombotic effects of sulfated polysaccharides in rabbits. Thromb Haemostas 1988; 60: 188-192
  PubMedGoogle Scholar
• 10 Ofosu FA, Gray E. Mechanisms of action of heparin: Applications to the development of derivatives of heparin and heparinoids with antithrombotic properties. Semin Thromb Hemostas 1988; 14: 9-17
  Article in Thieme ConnectPubMedGoogle Scholar
• 11 Abbadini M, Jin Zhu G, Maggi A, Pangrazzi J, Donati MB, Mussoni L. Dermatan sulphate induces plasminogen activator release in the perfused rat hindquarters. Blood 1987; 70: 1858-1860
  PubMedGoogle Scholar
• 12 Mascellani G, Bianchini P. European patent 1990, No. 0221977; USA patent 1990, No. 4973580.
  PubMed
• 13 Reyers I, Mysliwiec M, Mussoni L, Donati MB. Stasis induced venous thrombosis. In: Standardization of Animal Models of Thrombosis. Breddin K, Zimmermann R. (eds). Schattauer: Stuttgart; 1983: 99-108
• 14 Dejana E, Villa S, de Gaetano G. Bleeding time in rats: a comparison of different experimental conditions. Thromb Haemostas 1982; 48: 108-111
  PubMedGoogle Scholar
• 15 Colucci M, Paramo JA, Collen D. Generation in plasma of a fast-acting inhibitor of plasminogen activator in response to endotoxin stimulation. J Clin Invest 1985; 75: 818-824
  CrossrefPubMedGoogle Scholar
• 16 Emeis JJ, Kooistra T. Interleukin 1 and lipopolysaccharide induce an inhibitor of tissue-type plasminogen activator in vivo and in cultured endothelial cells. J Exp Med 1986; 163: 1260-1266
  CrossrefPubMedGoogle Scholar
• 17 Colucci M, Paramo JA, Collen D. Inhibition of one-chain and two-chain forms of human tissue-type plasminogen activator by the fast-acting inhibitor of plasminogen activator in vitro and in vivo. J Lab Clin Med 1986; 108: 53-59
  PubMedGoogle Scholar
• 18 de Fouw NJ, de Jong YF, Haverkate F, Bertina AM. The influence of thrombin and platelets on fibrin clot lysis rates in vitro: a study using a clot lysis system consisting of purified human proteins. Fibrinolysis 1988; 2: 235-244
  PubMedGoogle Scholar
• 19 Yin ET, Wessler S, Butler J. Plasma heparin: a unique, practical submicrogram sensitive assay. J Lab Clin Med 1973; 81: 298-310
  PubMedGoogle Scholar
• 20 Haverkate F, Brakman P. Fibrin plate assay. In: Progress in Chemical Fibrinolysis and Thrombolysis, Voll. Davidson JF, Samama MM, Desnoyers PC. (eds). Raven Press; New York: 1975: 151-159
• 21 Bergonzini GL, Bianchini P, Hoppensteadt D, Parma B, Osima B, Walenga JM. Fareed J. Pharmacokinetics of native and low molecular weight dermatans: preliminary studies in rats and primates. Semin Thromb Hemostas 1991; 17 (Suppl. 12) 235-9
  PubMedGoogle Scholar
• 22 Bjornsson TD, Levy G. Pharmacokinetics of heparin. I. Studies of dose dependence in rats. J Pharmacol Exp Ther 1979; 210: 237-242
  PubMedGoogle Scholar
• 23 Markwardt F. Past, present and future of hirudin. Haemostasis 1991; 21 Suppll 11-26
  PubMedGoogle Scholar
• 24 Marsh NA. Does heparin stimulate fibrinolysis?. Br J Haematol 1990; 76: 163-167
  CrossrefPubMedGoogle Scholar
• 25 Stein PL, van Zonneveld AJ, Pannekoek H, Strickland S. Structural domains of human tissue-type plasminogen activator that confer stimulation by heparin. J Biol Chem 1989; 264: 15441-15444
  PubMedGoogle Scholar