Effects of Heparin and Related Sulfated Polysaccharides on Tissue Factor Expression Induced by Mitogenic and Non-mitogenic Factors in Human Vascular Smooth Muscle Cells

Summary

Smooth muscle cells (SMCs) of the intima are generally quiescent and non proliferative. Their proliferation due to different stimulations occurs in myointimal hyperplasia and is regularly present in athero-genesis or after transluminal angioplasty leading to vascular occlusive stenosis. In the course of these pathologies, the Tissue Factor (TF) synthesis was upregulated and rapidly expressed at the membrane of the SMCs. Heparin is known to inhibit SMCs proliferation induced by FCS. We evaluated the inhibitory effect of heparin on the expression of TF induced by various mitogenic (FCS, PDGF-BB and EGF) and non-mitogenic (bacterial LPS) agents. Inhibition by heparin of SMCs proliferation induced by the same agonists was also determined.

Quiescent human vascular SMCs from normal adult arteries were treated for 1 h by heparin and related sulfated polysaccharides before stimulation by the agonists. All the agonists up-regulated the expression of TF antigen and activity. TF expression induced by the growth factors was inhibited by heparin (IC 50: 10-30 μg/ml), and other sulfated polysaccharides (IC 50: 1-5 μg/ml). SMCs proliferation, late activation of the extracellular signal-regulated kinases (ERK1/2), and PKC activity were inhibited by heparin (IC 50: 30-50 μg/ml) in SMCs stimulated by FCS but not in SMCs treated by PDGF or EGF. In contrast, heparin had no effect on LPS-induced TF expression nor on LPS-induced PKC activation. These results indicate that, besides its well known effect on SMC proliferation, heparin displays an inhibitory effect on cell mediated blood clotting processes through regulation of the TF expression.

• References

• 1 Zur M, Nemerson Y. Tissue Factor pathways of blood coagulation. Haemostasis and Thombosis. Bloom AL, and Thomas DP. (eds) Churchill Livingston – Edinburgh 1987; 148-63.
• 2 Drake TA, Morrissey JH, Edgington TS. Selective cellular expression of tissue factor in human tissues. Am J Pathol 1989; 134: 1087-97.
• 3 Wilcox JN, Smith KM, Schwartz SM, Gordon D. Localization of tissue factor in the normal vessel wall and in the atherosclerotic plaque. Proc Natl Acad Sci USA 1989; 86: 2839-43.
• 4 Toschi V, Gallo R, Lettino M, Fallon JT, Gertz SD, Fernández-Ortiz A, Chesebro JH, Badimon L, Nemerson Y, Fuster V, Badimon JJ. Tissue Factor modulates the thrombogenicity of human atherosclerotic plaques. Circulation 1997; 95: 594-9.
• 5 Thiruvikraman SV, Guha A, Roboz J, Taubman MB, Nemerson Y, Fallon JT. In situ localization of tissue factor in human artherosclerotic plaques by binding of digoxigenin-labeled factors VIIa and X. Lab Invest 1996; 75: 451-61.
• 6 Taubman MB, Marmur JD, Rosenfield CL, Guha A, Nichtberger S, Nemerson Y. Agonist-mediated tissue factor expression in cultured vascular smooth muscle cells. Role of Ca²⁺⁺ mobilization and protein kinace C activation. J Clin Invest 1993; 91: 547-52.
• 7 Tremoli E, Prati L, Parolari A, Antona C, Colli S, Mussoni L. Serum and thrombin induce tissue factor expression in human smooth muscle cells. Thromb Haemost (Abstract) 1995; 73: 1080
• 8 Xuereb JM, Sie P, Boneu B, Constans J. Up-regulation of tissue factor expression by platelet-derived growth factor in human vascular smooth muscle cells in culture. Thromb Haemost 1997; 78: 1520-6.
• 9 Ross R. The pathogenesis of atherosclerosis: a perspective for the 1990s. Nature 1993; 362: 801-9.
• 10 Sanders M. Molecular and cellular concepts in atherosclorosis. Pharmac Ther 1994; 61: 109-53.
• 11 Clowes AW Karnovsky MJ. Suppression by heparin of smooth muscle proliferation in injured arteries. Nature 1997; 265: 625-6.
• 12 Clowes AW, Clowes MM. Kinetics of cellular proliferation after arterial injury. II. Inhibition of smooth muscle growth by heparin. Lab Invest 1985; 52: 611-6.
• 13 Kimura I, Nagaura T, Naitoh T, Kobayashi S, Kimura M. Heparin inhibits the progression phase of subcultured endothelial cell proliferation in rat aorta. Japan J Pharmacol 1992; 60: 369-75.
• 14 Kilfeather SA Tagoe S, Perez AC Okona-Mensa K, Matin R, Page CP. Inhibition of serum-induced proliferation of bovine tracheal smooth muscle cells in culture by heparin and related glycosaminoglycans. Br J Pharmacol 1995; 114: 1442-6.
• 15 Karnovsky MJ, Wright TC, Castellot JJ, Choay J, Lormeau JC, Petitou M. Heparin, heparan sulfate, smooth muscle cells and atherosclerosis. Heparin and Related Polysaccharides. Structure and activities Annals New York Academy of Sciences 1989; 556: 268-81.
• 16 Wright TC, Pukac LA, Castellot JJ, Karnovsky MJ, Levine RA, Kim-Park HY, Campisi J. Heparin suppresses the induction of c-fos and c-myc mRNA in murine fibroblasts by selective inhibition of a protein kinase C-dependent pathway. Proc Natl Acad Sci USA 1989; 86: 3199-203.
• 17 Pukac LA, Castellot JJ, Wright TC, Caleb BL, Karnovsky MJ. Heparin inhibits c-fos and c-myc mRNA expression in vascular smooth muscle cells. Cell Regul 1990; 1: 435-43.
• 18 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.
• 19 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.
• 20 Herbert JM, Clowes M, Lea HJ, Pascal M, Clowes AW. Protein kinase C α expression is required for heparin inhibition of rat smooth muscle cell proliferation in vitro and in vivo. J Biol Chem 1996; 271 (42) 25928-35.
• 21 Ottlinger ME, Pukac LA, Karnovsky MH. Heparin inhibits mitogen-activated protein kinase activation in intact rat vascular smooth muscle cells. J Biol Chem 1993; 268: 19173-6.
• 22 Pukac LA, Carter JE, Ottlinger ME, Karnovsky MJ. Mechanisms of inhibition by heparin of PDGF stimulated MAP kinase activation in vascular smooth muscle cells. J Cell Physiol 1997; 172: 69-78.
• 23 Daum G, Hedin U, Wang Y, Wang T, Clowes AW. Diverse effects of heparin on mitogen-activated protein kinase-dependent signal transduction in vascular smooth muscle cells. Circulation Res 1997; 81: 17-23.
• 24 Gotoh Y., Nishida E, Yamashita T, Hoshi M, Kawakami M. Sakai H Micro-tubule-associated protein (MAP)kinase activated by nerve growth factor and epidermal growth factor in PC 12 cells. Identity with the MAPK of fibroblastic cells. Eur J Biochem 1991; 10: 2661-8.
• 25 McCaffey PG, Friedman B, Rosner MR. Diaglycerol modulates binding and phosphorylation of the epidermal growth factor receptor. J Biol Chem 1984; 259: 12502-7.
• 26 Almus FE, Rao LVM, Rapaport I. S. Decreased inducibility of tissue factor activity on human umbilical vein endothelial cells cultured with endothelial cell growth factor and heparin. Thromb Res 1988; 50: 339-44.
• 27 Almus FE, Rao LVM, Pendurthi UR, Quattrochi L, Rapaport SI. Mechanism for diminished Tissue Factor expression by endothelial cells cultured with heparin binding growth factor-1 and heparin. Blood 1991; 6 (77) 1256-62.
• 28 Cadroy Y, Gaspin D, Dupouy D, Lormeau JC, Boneu B, Sié P. Heparin reverses the procoagulant properties of stimulated endothelial cells. Thromb Haemost 1996; 75 (01) 190-5.
• 29 Pepe G, Giusti B, Attanasio M, Gori AM, Comeglio P, Martini F, Gensini G, Abbate R, Neri Serneri GG. Tissue factor and plasminogen activator inhibitor type 2 expression in human stimulated monocytes is inhibited by heparin. Sem Thromb Haemost 1997; 23: 135-41.
• 30 Kahan C, Seuwen K, Meloche S, Pouyssegur J. Coordinate, biphasic activation of p44 mitogen-activated protein kinase and S6 kinase by growth factors in hamster fibroblasts. J Biol Chem 1992; 267: 13369-75.
• 31 Lenormand P, Sardet C, Pages G, L'allemain G, Brunet A, Pouyssegur J. Growth factors induce nuclear translocation of MAP kinases (p42^MAPK and p44^MAPK) but not of their activator MAP kinases (p45^MAPK) in fibroblasts. J Cell Biol 1993; 122: 1079-88.
• 32 Langan EM, Youkey JR, Elmore JR, Franklin DP, Singer HA. Regulation of MAP kinase activity by growth stimuli in vascular smooth muscle. J Surg Res 1994; 57: 215-20.
• 33 Kelleher MD, Abe MK, Chao T, Jain M, Green J, Solway J, Rosner MR, Hershenson MB. Role of MAP kinase activation in bovine tracheal smooth muscle mitogenesis. Am J Physiol 1995; 268: L894-901.
• 34 Mackman N, Brand K, Edgington TS. Liposaccharide mediated transcriptional activation of the human tissue factor gene in THP-1 monocytic cells require both activator protein 1 and nuclear factor κB binding site. J Exp Med 1991; 174: 1517-26.
• 35 Ollivier V, Houssaye S, Ternisien C, Léon A, De Verneuil H, Elbim C, Mackman N, Edgington TS, de Prost D. Endotoxin-induced tissue factor messenger RNA in human monocytes is negatively regulated by a cyclic AMP-dependent mechanism. Blood 1993; 81: 973-9.
• 36 Oeth P, Parry GCN, Mackman N. Regulation of the tissue factor gene in human monocytic cells. Arterioscler Thromb Vasc Biol 1997; 17: 365-74.
• 37 Osnes LTN, Foss KB, Joo GB, Okkenhaug C, Westvik AB, Ovstebo R, Kierulf P. Acetylsalicylic acid and sodium salicylate inhibit LPS-induced NF-κB/c-rel nuclear translocation, and synthesis of tissue factor (TF) and tumor necrosis factor alfa (TNF-α) in human monocytes. Thromb Haemost 1996; 76: 970-6.
• 38 Moll T, Malgorzata C, Holzmuller H, Hofer-Warbinek R, Wagner E, Wink-ler H, Bach FH, Hofer E. Regulation of the tissue factor promoter in endothelial cells. J Biol Chem 1995; 270: 3849-57.
• 39 Parry GCN, Mackman N. Transcriptional regulation of tissue factor expression in human endothelial cells. Arterioscler Thromb Vasc Biol 1995; 15: 612-21.
• 40 Ollivier V, Parry GCN, Cobb R, de Prost D, Mackman N. Elevated cyclic AMP inhibits NF-κB mediated transcription in human monocytic cells and endothelial cells. J Biol Chem 1996; 271: 20828-35.
• 41 Mackman N. Regulation of the tissue factor gene. Thromb Haemost 1997; 78: 747-54.
• 42 Taby O, Rosenfield CL, Bogdanov V, Nemerson Y, Taubman MB. Cloning of the rat tissue factor cDNA and promoter: identification of a serum-response region. Thromb Haemost 1996; 76: 697-702.
• 43 Sato Y, Asada Y, Marutsuka K, Hatakeyama K, Sumiyoshi A. Tissue factor induces migration of cultured aortic smooth muscle cells. Thromb Haemost 1996; 75: 389-92.