The Antimitogenic Action of the Sulphated Polysaccharide Fucoidan Differs from Heparin in Human Vascular Smooth Muscle Cells

Summary

The sulphated polysaccharides fucoidan and heparin both inhibit vascular smooth muscle cell (VSMC) proliferation. In this study we compared their actions on mitogenesis and ERK1/ERK2 activation in human VSMC. Although they displaced cell surface [3H]-heparin binding with similar affinity, they exerted clearly distinguishable actions. Fucoidan potently inhibited DNA synthesis stimulated by foetal calf serum, PDGF-BB and thrombospondin-1. Heparin inhibited the mitogenic action of serum and thrombospondin-1 (though less potently than fucoidan), but failed to inhibit PDGF-BB-induced DNA synthesis. In parallel studies, fucoidan, but not heparin, inhibited ERK1/ERK2 activation by PDGF-BB. Moreover, fucoidan inhibited serum-induced mitogenesis in “heparin resistant” VSMC, which are refractory to heparin’s antimitogenic action. In summary, the structurally different polysaccharides, heparin, fucoidan (and fucans) have distinguishable effects on mitogenesis and ERK1/ERK2 activation, suggesting that different mechanism(s) mediate these actions. The potent antimitogenic action of fucoidan and its efficacy in heparin resistant VSMC emphasise the need to further investigate its mechanism of action in human VSMC and suggest this agent could have therapeutic potential.

Keywords

Fucoidan - heparin - human vascular smooth muscle - DNA synthesis - ERK1/ERK2 activation

• References

• 1 Casu B. Structure and biological activity of heparin. Adv Carbohydr Chem Biochem 1985; 43: 51-134.
• 2 Boisson-Vidal C, Haroun F, Ellouali M, Blondin C, Fischer AM, De Agostini A, Josefonvicz J. Biological activities of polysaccharides from marine algae. Drugs Fut 1995; 20: 1237-49.
• 3 Pereira MS, Mulloy B, Mourao PA. Structure and anticoagulant activity of sulfated fucans. Comparison between the regular, repetitive, and linear fucans from echinoderms with the more heterogeneous and branched polymers from brown algae. J Biol Chem 1999; 274: 7656-67.
• 4 Chizhov AO, Dell A, Morris HR, Haslam SM, McDowell RA, Shashkov AS, Nifant’ev NE, Khatuntseva EA, Usov AI. A study of fucoidan from the brown seaweed Chorda filum. Carbohydr Res 1999; 320: 108-19.
• 5 Chevolot B, Mulloy B, Ratiskol J, Foucault A, Colliec-Jouault S. A disaccharide repeat unit is the major structure in fucoidans from two species of brown algae. Carbohydr Res 2001; 330: 529-35.
• 6 Church FC, Meade JB, Treanor RE, Whinna HC. Antithrombin activity of fucoidan. The interaction of fucoidan with heparin cofactor II, antithrombin III, and thrombin. J Biol Chem 1989; 264: 3618-23.
• 7 Guyton JR, Rosenberg RD, Clowes AW, Karnovsky MJ. Inhibition of rat arterial smooth muscle cell proliferation by heparin. In vivo studies with anticoagulant and nonanticoagulant heparin. Circ Res 1980; 46: 625-34.
• 8 Castellot JJ, Wong K, Herman B, Hoover RL, Albertini DF, Wright TC, Caleb BL, Karnovsky MJ. Binding and internalization of heparin by vascular smooth muscle cells. J Cell Physiol 1985; 124: 13-20.
• 9 Logeart D, Prigent-Richard S, Jozefonvicz J, Letourneur D. Fucans, sulfated polysaccharides extracted from brown seaweeds, inhibit vascular smooth muscle cell proliferation. I. Comparison with heparin for antiproliferative activity, binding and internalization. Eur J Cell Biol 1997; 74: 376-84.
• 10 Logeart D, Prigent-Richard S, Boisson-Vidal C, Chaubet F, Durand P, Jozefonvicz J, Letourneur D. Fucans, sulfated polysaccharides extracted from brown seaweeds, inhibit vascular smooth muscle cell proliferation. II. Degradation and molecular weight effect. Eur J Cell Biol 1997; 74: 385-90.
• 11 Letourneur D, Caleb BL, Castellot JJ. Heparin binding, internalization, and metabolism in vascular smooth muscle cells: II. Degradation and secretion in sensitive and resistant cells. J Cell Physiol 1995; 165: 687-95.
• 12 Clowes AW, Karnowsky MJ. Suppression by heparin of smooth muscle cell proliferation in injured arteries. Nature 1977; 265: 625-6.
• 13 Hoover RL, Rosenberg R, Haering W, Karnovsky MJ. Inhibition of rat arterial smooth muscle cell proliferation by heparin. II. In vitro studies. Circ Res 1980; 47: 578-83.
• 14 Majack RA, Clowes AW. Inhibition of vascular smooth muscle cell migration by heparin-like glycosaminoglycans. J Cell Physiol 1984; 118: 253-6.
• 15 Majack RA, Cook SC, Bornstein P. Platelet-derived growth factor and heparin-like glycosaminoglycans regulate thrombospondin synthesis and deposition in the matrix by smooth muscle cells. J Cell Biol 1985; 101: 1059-70.
• 16 Snow AD, Bolender RP, Wight TN, Clowes AW. Heparin modulates the composition of the extracellular matrix domain surrounding arterial smooth muscle cells. Am J Pathol 1990; 137: 313-30.
• 17 Au YP, Montgomery KF, Clowes AW. Heparin inhibits collagenase gene expression mediated by phorbol ester-responsive element in primate arterial smooth muscle cells. Circ Res 1992; 70: 1062-9.
• 18 Clowes AW, Clowes MM, Kirkman TR, Jackson CL, Au YP, Kenagy R. Heparin inhibits the expression of tissue-type plasminogen activator by smooth muscle cells in injured rat carotid artery. Circ Res 1992; 70: 1128-36.
• 19 Kenagy RD, Nikkari ST, Welgus HG, Clowes AW. Heparin inhibits the induction of three matrix metalloproteinases (stromelysin, 92-kD gelatinase, and collagenase) in primate arterial smooth muscle cells. J Clin Invest 1994; 93: 1987-93.
• 20 Reilly CF, Kindy MS, Brown KE, Rosenberg RD, Sonenshein GE. Heparin prevents vascular smooth muscle cell progression through the G1 phase of the cell cycle. J Biol Chem 1989; 264: 6990-5.
• 21 Pukac LA, Ottlinger ME, Karnovsky MJ. Heparin suppresses specific second messenger pathways for protooncogene expression in rat vascular smooth muscle cells. J Biol Chem 1992; 267: 3707-11.
• 22 Castellot JJ, Pukac LA, Caleb BL, Wright TC, Karnovsky MJ. Heparin selectively inhibits a protein kinase C-dependent mechanism of cell cycle progression in calf aortic smooth muscle cells. J Cell Biol 1989; 109: 3147-55.
• 23 Ottlinger ME, Pukac LA, Karnovsky MJ. Heparin inhibits mitogen-activated protein kinase activation in intact rat vascular smooth muscle cells. J Biol Chem 1993; 268: 19173-6.
• 24 Delmolino LM, Castellot JJ. Heparin suppresses sgk, an early response gene in proliferating vascular smooth muscle cells. J Cell Physiol 1997; 173: 371-9.
• 25 Mishra-Gorur K, Castellot JJ. Heparin rapidly and selectively regulates protein tyrosine phosphorylation in vascular smooth muscle cells. J Cell Physiol 1999; 178: 205-15.
• 26 Lindner V, Olson NE, Clowes AW, Reidy MA. Inhibition of smooth muscle cell proliferation in injured rat arteries. Interaction of heparin with basic fibroblast growth factor. J Clin Invest 1992; 90: 2044-9.
• 27 Besner G, Higashiyama S, Klagsbrun M. Isolation and characterization of a macrophage-derived heparin-binding growth factor. Cell Regul 1990; 01: 811-9.
• 28 Fager G, Camejo G, Bondjers G. Heparin-like glycosaminoglycans influence growth and phenotype of human arterial smooth muscle cells in vitro. I. Evidence for reversible binding and inactivation of the platelet-derived growth factor by heparin. In Vitro Cell Dev Biol 1992; 28A: 168-75.
• 29 Sperinde GV, Nugent MA. Heparan sulfate proteoglycans control intracellular processing of bFGF in vascular smooth muscle cells. Biochemistry 1998; 37: 13153-64.
• 30 McCaffrey TA, Falcone DJ, Brayton CF, Agarwal LA, Welt FG, Weksler BB. Transforming growth factor-beta activity is potentiated by heparin via dissociation of the transforming growth factor-beta/alpha 2-macroglobulin inactive complex. J Cell Biol 1989; 109: 441-8.
• 31 Hughes AD, Patel M, Wijetunge S, Clunn G, Schachter M. Comparison of effects of platelet-derived growth factor isoforms on signaling and DNA synthesis of human cultured saphenous vein cells. J Cardiovasc Pharmacol 1995; 25: 481-5.
• 32 Patel MK, Lymn JS, Clunn GF, Hughes AD. Thrombospondin-1 is a potent mitogen and chemoattractant for human vascular smooth muscle cells. Arterioscler Thromb Vasc Biol 1997; 17: 2107-14.
• 33 Patel MK, Refson JS, Schachter M, Hughes AD. Characterisation of [3H]-heparin binding in human vascular smooth muscle cells and its relationship to the inhibition of DNA synthesis. Br J Pharmacol 1999; 127: 361-8.
• 34 Chan P, Patel M, Betteridge L, Munro E, Schachter M, Wolfe J, Sever P. Abnormal growth regulation of vascular smooth muscle cells by heparin in patients with restenosis. Lancet 1993; 341: 341-2.
• 35 Refson JS, Schachter M, Patel MK, Hughes AD, Munro E, Chan P, Wolfe JH, Sever PS. Vein graft stenosis and the heparin responsiveness of human vascular smooth muscle cells. Circulation 1998; 97: 2506-10.
• 36 Chan P, Munro E, Patel M, Betteridge L, Schachter M, Sever P, Wolfe J. Cellular biology of human intimal hyperplastic stenosis. Eur J Vasc Surg 1993; 07: 129-35.
• 37 Patel MK, Betteridge LJ, Hughes AD, Clunn GF, Schachter M, Shaw RJ, Sever PS. Effect of angiotensin II on the expression of the early growth response gene c-fos and DNA synthesis in human vascular smooth muscle cells. J Hypertens 1996; 14: 341-7.
• 38 Cheng Y, Prusoff WH. Relationship between the inhibition constant (K_i) and the concentration of inhibitor which causes 50 per cent inhibition (I₅₀) of an enzymatic reaction. Biochem Pharmacol 1973; 22: 3099-108.
• 39 Vischer P, Buddecke E. Different action of heparin and fucoidan on arterial smooth muscle cell proliferation and thrombospondin and fibronectin metabolism. Eur J Cell Biol 1991; 56: 407-14.
• 40 McCaffrey TA, Falcone DJ, Borth W, Brayton CF, Weksler BB. Fucoidan is a non-anticoagulant inhibitor of intimal hyperplasia. Biochem Biophys Res Commun 1992; 184: 773-81.
• 41 Religa P, Kazi M, Thyberg J, Gaciong Z, Swedenborg J, Hedin U. Fucoidan inhibits smooth muscle cell proliferation and reduces mitogen-activated protein kinase activity. Eur J Vasc Endovasc Surg 2000; 20: 419-26.
• 42 Castellot JJ, Cochran DL, Karnovsky MJ. Effect of heparin on vascular smooth muscle cells. I. Cell metabolism. J Cell Physiol 1985; 124: 21-8.
• 43 Hedin U, Daum G, Clowes AW. Heparin inhibits thrombin-induced mitogen-activated protein kinase signaling in arterial smooth muscle cells. J Vasc Surg 1998; 27: 512-20.