Advertisement

ANTICOAGULANT FUCOIDAN FRACTIONS FROM FUCUS VESICULOSUS INDUCE PLATELET ACTIVATION IN VITRO

      Abstract

      Anticoagulant fucoidan fractions of different molecular weight and sulfate content were prepared and investigated for their effects on platelet function in vitro. The fucoidan fractions were incubated with human platelet rich plasma (PRP) at concentrations of 5, 10 and 50 μg/ml. Platelet activation was subsequently studied by a standard aggregation assay and flow cytometric determination of the activation dependent platelet-surface markers CD62p (P-selectin, GMP-140) and CD63 (GP53). All fucoidan fractions induced irreversible platelet aggregation in a dose-dependent manner. Comparing fractions of identical molecular weight (100 kDa) the low sulfate content fucoidan FF5 (S=7,6%) exerted a significantly greater effect than the highly sulfated fucoidan FF7 (S= 10,2%) over the whole concentration range (n=5, P<0,05). Among fractions of identical sulfate content fucoidan-induced platelet aggregation was also found to depend on the molecular weight of the fucoidan. At concentrations of 10 and 50 μg/ml the high molecular weight fraction FF7/1 (150 kDa) showed a significantly greater effect than the 50 kDa fraction FF7/3 (24,8 ± 6,7 vs. 7,0 ± 3,5 and 54,6 ± 13,5 vs. 15,0 ± 9,0%, respectively; mean ± SD, n=5, P<0,05). The molecular weight dependence of the fucoidan effect was also reflected by the flow cytometric data. Coincubation of FF7/1 and FF7/3 (10μg/ml) with PRP increased the number of CD62p and CD63 positive platelets by 9,0 ± 3,3 vs. 2 ± 1,9 and 7,1 ± 2,4 vs. 3,2 ± 2,6% over control values, respectively (n=5, P< 0,05). In conclusion, our results show that the low molecular weight fucoidan FF7/3 combines potent anticoagulant and fibrinolytic properties with only minor platelet activating effects and is therefore a suitable substance for further pharmacological studies. © 1997 Elsevier Science Ltd

      Keywords

      To read this article in full you will need to make a payment

      Purchase one-time access:

      Academic & Personal: 24 hour online accessCorporate R&D Professionals: 24 hour online access
      One-time access price info
      • For academic or personal research use, select 'Academic and Personal'
      • For corporate R&D use, select 'Corporate R&D Professionals'

      Subscribe:

      Subscribe to Thrombosis Research
      Already a print subscriber? Claim online access
      Already an online subscriber? Sign in
      Institutional Access: Sign in to ScienceDirect

      References

        • SPINGER G.F
        • WURZEL H.A
        • McNEAL Jr., G.M
        • ANSELL N.J
        • DOUGHTY M.F
        Isolation of anticoagulant fractions from crude fucoidan.
        Proc. Soc. Exp. Biol. Med. 1957; 94: 404-409
        • GRAUFFEL V
        • KLOAREG B
        • MABEAU S
        • DURAND P
        • JOZEFONVICZ J
        New natural polysaccharides with potent antithrombic activity: fucans from brown algae.
        Biomaterials. 1989; 10: 363-368
        • CHURCH F.C
        • MEADE J.B
        • TREANOR R.E
        • WHINNA H.C
        Antithrombin activity of fucoidan.
        J. Biol. Chem. 1989; 264: 3618-3623
        • SOEDA S
        • SAKAGUCHI S
        • SHIMENO H
        • NAGAMATSU A
        Fibrinolytic and anticoagulant activities of highly sulfated fucoidan.
        Biochem. Pharmacol. 1992; 43: 1853-1858
        • SOEDA S
        • OHMAGURI Y
        • SHIMENO H
        • NAGAMATSU A
        Preparation of oversulfated fucoidan fragments and evaluation of their antithrombotic activities.
        Thromb. Res. 1993; 72: 247-256
        • McCAFFREY T.A
        • FALCONE
        • BORTH W
        • BRAYTON C.F
        • WEKSLER B.B
        Fucoidan is a novel non-anticoagulant inhibitor of intimal hyperplasia.
        Biochem. Biophys. Res. Commun. 1992; 184: 773-781
        • FUSTER V
        • FALK E
        • FALLON J.T
        • BADIMON L
        • CHESEBRO J.H
        • BADIMON J.J
        The three processes leading to post PTCA restenosis: dependence on the lesion substrate.
        Thromb. Haemostasis. 1995; 74: 552-559
        • MARUYAMA H
        • NAKAJIMA J
        • YAMAMOTO I
        A study on the anticoagulant and fibrinolytic activities of a crude fucoidan from the edible brown seaweed Laminaria religiosa, with special reference to its inhibitory effect on the growth of sarcoma-180 ascites cells subcutaneously implanted into mice.
        Kitasato Arch. of Exp. Med. 1987; 60: 105-121
      1. BRUHN, T., DÜRIG, J., KRAISELBURD, E.N., DE CLERCQ, BRUHN, H.D., AND BÉRESS, L. Antiviral and anticoagulant activity of polysaccharides from marine brown algae. In: Biochemical Aspects of Marine Pharmacology. Eds.: Lazarovici, P., Spira, M.E., Zlotkin, E., Alaken Inc., Fort Collins, Colorado, 187–208, 1996.

        • FISCHER A.M
        • MAUZAC M
        • TAPON-BRETAUDIERE J
        • JOZEFOWICZ J
        Anticoagulant activity of dextran derivatives..
        Biomaterials. 1985; 6: 198-202
        • NAITO K
        • AOKI N
        Assay of antiplasmin α 2-activity by means of a plasmin specific peptide substrate.
        Thromb. Res. 1978; 12: 1147-1156
        • BORN G.V.R
        Quantitative investigations into the aggregation of blood platelets.
        J. Physiol. 1962; 162: 927-929
        • PATANKAR M.S
        • OEHNINGER S
        • BARNETT T
        • WILLIAMS R.L
        • CLARK G.F
        A revised structure for fucoidan may explain some of its biological activities.
        J. Biol. Chem. 1993; 268: 21770-21776
        • SALZMANN E.W
        • ROSENBERG R.D
        • SMITH M.H
        • LINDON J.N
        • FAVREAU L
        Effect of heparin and heparin fractions on platelet aggregation.
        J. Clin. Invest. 1980; 65: 64-73
        • METZELAAR M.J
        • SIXMA J.J
        • NIEUWENHUIS H.K
        Detection of platelet activation using specific monoclonal antibodies.
        Blood Cells. 1990; 16: 85-96
        • WUN T
        • PAGLIERONI T.G
        • LACHANT N.A
        Desmopressin stimulates expression of P-selectin on human platelets in vitro.
        J. Lab. Clin. Med. 1995; 126: 401-409
        • OLEKSOWICZ L
        • MROWIEC Z
        • ISAACS R
        • DUTCHER I.P
        • PUSZKIN E
        Morphologic and ultrastructural evidence of interleukin-6 induced platelet activation.
        Am. J. Hematol. 1995; 48: 92-99
        • KANNAN K
        • DIVERS S.G
        • LURIE A.A
        • CHERVENAK R
        • FUKUDA M
        • HOLCOMBE R.F
        Cell surface expression of lysosome-associated membrane protein-2 (lamp2) and CD63 as markers of in vivo platelet activation in malignancy.
        Eur. J. Haematol. 1995; 55: 145-151
        • TSCHOEPE D
        • SCHULTHEISS H.P
        • KOLAROV P
        • SCHWIPPERT B
        • DANNEHL K
        • NIEUWENHUIS H.K
        • KEHREL B
        • STRAUER B
        • GRIES F.A
        Platelet membrane activation markers are predictive for increased risk of ischemic events after PTCA.
        Circulation. 1993; 88: 37-42
        • SHATTIL S
        • CUNNINGHAM M
        • HOXIE J.A
        Detection of activated platelets in whole blood using activation-dependent monoclonal antibodies and flow cytometry.
        Blood. 1988; 70: 307-315
        • LONGBIN L
        • FREEDMAN J
        • HORNSTEIN A
        • FENTON J.W
        • OFOSU F.A
        Thrombin binding to platelets and their activation in plasma.
        Br. J. Haematol. 1994; 88: 592-600
        • CHONG B.H
        Heparin-induced thrombocytopenia.
        Blood Rev. 1988; 2: 108-114
        • HORNE M.K
        • CHAO E.S
        Heparin binding to resting and activated platelets.
        Blood. 1989; 74: 238-243
        • TARABOLETTI G
        • ROBERTS D
        • LIOTTA A
        • GIAVAZZI R
        Platelet thrombospondin modulates endothelial cell adhesion, motility, and growth: a potential angiogenesis regulatory factor.
        J. Cell. Biol. 1990; 111: 765-772
        • KAZUKO H
        • NUDELMAN D
        • STROUD M.R
        • SHIOZAWA T
        • HAKOMORI S
        Selectin GMP-140 (CD62; PADGEM) binds to sialosyl-Lea and sialosyl-Lex, and sulfated glycans modulate this binding.
        Biochem. Biophys. Res. Commun. 1991; 181: 1223-1230
      2. GUTENSOHN, K., BRUHN, T., KÖSTER, R., DÜRIG, J., KAEHLER, J., ABU, J., AND KUEHNL, P. Effects of fucoidan fractions on platelet antigens and proliferation of smooth muscle cells. Abstract, 27. Hämophilie-Symposium, Hamburg, 1996.