Anticoagulant fucoidan fractions from Fucus vesiculosus induce platelet activation in vitro.
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 micrograms/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 micrograms/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 (10 micrograms/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.
Dürig
J
J
Dept. of Internal Medicine, University Hospital Kiel, Germany.
Bruhn
T
T
Zurborn
K H
KH
Gutensohn
K
K
Bruhn
H D
HD
Béress
L
L
D000925:Anticoagulants / Q000494:pharmacology*; D015703:Antigens, CD / Q000032:analysis; D015815:Cell Adhesion Molecules / Q000032:analysis; D005342:Fibrinolysis / Q000187:drug effects; D005434:Flow Cytometry; D008401:Gas Chromatography-Mass Spectrometry; D006801:Humans; D066298:In Vitro Techniques; D008297:Male; D008970:Molecular Weight; D009391:Nephelometry and Turbidimetry; D019007:P-Selectin / Q000032:analysis; D000459:Phaeophyceae / Q000737:chemistry; D015539:Platelet Activation / Q000187:drug effects*; D010974:Platelet Aggregation; D010978:Platelet Factor 4 / Q000032:analysis; D010980:Platelet Membrane Glycoproteins / Q000032:analysis; D011134:Polysaccharides / Q000737:chemistry / Q000494:pharmacology*; D013329:Structure-Activity Relationship; D013431:Sulfates / Q000032:analysis; D013463:Sulfuric Acid Esters / Q000737:chemistry / Q000494:pharmacology*; D060149:Tetraspanin 30
D000925:Anticoagulants; D015703:Antigens, CD; C554586:CD63 protein, human; D015815:Cell Adhesion Molecules; D019007:P-Selectin; D010980:Platelet Membrane Glycoproteins; D011134:Polysaccharides; D013431:Sulfates; D013463:Sulfuric Acid Esters; D060149:Tetraspanin 30; D010978:Platelet Factor 4; C007789:fucoidan