Effects of Heparin and Endothelial Cell Growth Supplement on Haemostatic Functions of Vascular Endothelium

Summary

Heparin in combination with endothelial cell growth factor (ECGF) affects physiological responses and growth of human umbilical vein endothelial cells (HUVEC). We have examined the effect of heparin, crude ECGF (endothelial cell growth supplement [ECGS]), or both on the basal and thrombin challenged output of metabolites by HUVEC. The supernatant and/or cell lysate was assayed for released prostacyclin, von Willebrand factor, tissue plasminogen activator, plasminogen activator inhibitor and thrombospondin. Heparin modified release of all these metabolites when in combination with ECGS, and in general these responses were the opposite of those generated by inflammatory mediators such as interleukin-1. It has been postulated that heparin acts by potentiating the effect of ECGF, but heparin inhibited thrombospondin release and enhanced that of von Willebrand factor in the absence of ECGS, while ECGS alone inhibited release of plasminogen activator inhibitor. Thus, under our experimental conditions it would appear that heparin and crude ECGF can affect HUVEC independently of one another.

• References

• 1 Thornton SC, Mueller SN, Levine EM. Human endothelial cells: use of heparin in cloning and long-term serial cultivation. Science 1983; 222: 623-625
• 2 Lobb RR, Harper JW, Fett JW. Purification of heparin-binding growth factors. Anal Biochem 1986; 154: 1-14
• 3 Maciag T, Mehlman T, Friesel R. Heparin binds endothelial cell growth factor, the principal endothelial cell mitogen in bovine brain. Science 1984; 225: 932-934
• 4 Burgess WH, Mehlman T, Marshak DR, Fraser BA, Maciag T. Structural evidence that endothelial cell growth factor p is the precursor of both endothelial cell growth factor a and acidic fibroblast growth factor. Proc Natl Acad Sci USA 1986; 83: 7216-7220
• 5 Barzu T, Lormeau JC, Petitou M, Michelson S, Choay J. Heparin-derived oligosaccharides: affinity for acidic fibroblast growth factor and effect on its growth-promoting activity for human endothelial cells. J Cell Physiol 1989; 140: 538-548
• 6 Damon DH, Lobb RR, D’Amore PA, Wagner JA. Heparin potentiates the action of acidic fibroblast growth factor by prolonging its biological half-life. J Cell Physiol 1989; 138: 221-226
• 7 Godspodarowicz D, Cheng J. Heparin protects basic and acidic fibroblast growth factor from inactivation. J Cell Physiol 1986; 128: 475-484
• 8 Schreiber AB, Kenney J, Kowalski WJ, Friesel R, Mehlman T, Maciag T. Interaction of endothelial cell growth factor with heparin: characterization by receptor and antibody recognition. Proc Natl Acad Sci USA 1985; 82: 6138-6142
• 9 Almus FE, Vijaya Mohan Rao L, 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; 77: 1256-1262
• 10 Andoh K, Petterson KS, Filion-Myklebust C, Prydz H. Observations on the cell biology of tissue factor in endothelial cells. Thromb Haemostas 1990; 63: 298-302
• 11 Weksler BB. Heparin and acidic fibroblast growth factor interact to decrease prostacyclin synthesis in human endothelial cells by affecting both prostacyclin H synthase and prostacyclin synthase. J Cell Physiol 1990; 142: 514-522
• 12 Boutherin-Falson O, Blaes N. Decreased prostacyclin production by human umbilical vein endothelial cells cultured with endothelial cell growth factor and heparin. Thromb Res 1989; 54: 487-492
• 13 Almus FE, Vijaya Mohan Rao L, Rapaport SI. Decreased inducibility of tissue factor activity on human umbilical vein endothelial cell growth factor and heparin. Thromb Res 1988; 50: 339-344
• 14 Konkle BA, Ginsburg D. The addition of endothelial cell growth factor and heparin to human umbilical vein endothelial cell cultures decreases plasminogen activator inhibitor-1 expression. J Clin Invest 1988; 82: 579-585
• 15 Maciag T, Cerundolo J, Ilsley S, Kelley PR, Forand R. An endothelial cell growth factor from bovine hypothalamus: identification and partial characterisation. Proc Natl Acad Sci USA 1979; 76: 5674-5678
• 16 Jaffe EA, Nachman RL, Becker CG, Minick CR. Culture of human endothelial cells derived from umbilical veins. Identification by morphologic and immunologic criteria. J Clin Invest 1973; 52: 2745-2756
• 17 McArthur MM, MacGregor IR, Prowse CV, Hunter NR, Dawes J, Pepper DS. The use of human endothelial cells cultured in flat wells and on microcarrier beads to assess tissue plasminogen activator and factor VIII related antigen release. Thromb Res 1986; 41: 581-587
• 18 Dawes J, Clemetson KJ, Gogstad GD, McGregor J, Clezardin P, Prowse CV, Pepper DS. A radioimmunoassay for thrombospondin, used in a comparative study of thrombospondin, p-thromboglobulin and platelet factor 4 in healthy volunteers. Thromb Res 1983; 29: 569-581
• 19 Hasegawa N, Yamamoto M, Yamamoto K. Stimulation of cell growth and inhibition of prostacyclin production by heparin in human umbilical vein endothelial cells. J Cell Physiol 1988; 137: 603-607
• 20 Itoh F, Kaji T, Hayakama Y, Oguma Y, Sakuragawa N. Heparin enhances thrombin-stimulated prostacyclin I₂ production by cultured endothelial cells. Thromb Res 1990; 57: 481-488
• 21 Ingerman-Wojenski CM, Silver MJ. Prostacyclin synthesis by endothelial cells from human umbilical veins: effect of cumulative population doublings. Prostaglandins 1988; 36: 127-137
• 22 Ristimaki A, Ylikorkala O, Viinikka L. Effect of growth factors on human vascular endothelial cell prostacyclin production. Arteriosclerosis 1990; 10: 653-657
• 23 Hanss M, Collen D. Secretion of tissue-type plasminogen activator and plasminogen activator inhibitor by cultured human endothelial cells: modulation by thrombin, endotoxin, and histamine. J Lab Clin Med 1987; 109: 97-104
• 24 Marsh NM, Minter AJ, Chesterman CN. The effect of heparin and other glycosaminoglycans on levels of tissue plasminogen activator and plasminogen activator inhibitor in cultured human umbilical vein endothelial cells. Blood Coag Fibrin 1990; 1: 133-138
• 25 Levin EG, Marzec V, Anderson J, Harker LA. Thrombin stimulates tissue plasminogen activator release from cultured endothelial cells. J Clin Invest 1984; 74: 1988-1995
• 26 Gelehrter TD, Sznycer-Laszuk R. Thrombin induction of plasminogen-activator-inhibitor in cultured human endothelial cells. J Clin Invest 1986; 77: 165-169
• 27 Konkle BA, Kollros PR, Kelly MD. Heparin-binding growth factor-1 modulation of plasminogen activator inhibitor-1 expression. J Biol Chem 1990; 265: 21867-21873
• 28 Lyons-Giordano B, Brinker JM, Kefalides NA. The effect of heparin on fibronectin and thrombospondin synthesis and mRNA levels in cultured human endothelial cells. Exp Cell Res 1990; 186: 39-46
• 29 Chesterman CN, Ager A, Gordon JL. Regulation of prostaglandin production and ectoenzyme activities in cultured aortic endothelial cells. J Cell Physiol 1983; 116: 45-50
• 30 Tan EM, Dodge GR, Sorger T, Kovalsky I, Unger GA, Yang L, Levine EM, Iozzo RV. Modulation of extracellular matrix gene expression by heparin and endothelial cell growth factor in human smooth muscle cells. Lab Invest 1991; 64: 474-482
• 31 Vlodavsky I, Folkman J, Sullivan R, Fridman R, Ishai-Michaeli R, Sasse J, Klagsbrun M. Endothelial cell-derived basic fibroblast growth factor: synthesis and deposition in subendothelial extracellular matrix. Proc Natl Acad Sci USA 1987; 84: 2292-2296
• 32 Mikhailides DP, Barradas MA, O’Donoghue S, Dandona P. Evidence for in vivo platelet activation following the injection of conventional unfractionated heparin. Platelets 1990; 1: 189-192
• 33 Marsh N. Does heparin stimulate fibrinolysis?. Br J Haematol 1990; 76: 163-167
• 34 Albrightson CR, Baenziger NL, Needleman P. Exaggerated human vascular cell prostaglandin biosynthesis mediated by monocytes: role of monokines and interleukin 1. J Immunol 1985; 135: 1872-1877
• 35 Rossi V, Breviario F, Ghezzi P, Dejana E, Mantovani A. Prostacyclin synthesis induced in vascular cells by interleukin-1. Science 1985; 229: 174-176
• 36 Nachman RL, Hajjar KA, Silverstein RL, Dinarello CA. Interleukin 1 induces endothelial cell synthesis of plasminogen activator inhibitor. J Exp Med 1986; 163: 1595-1600
• 37 Bevilacqua MP, Schleef RR, Gimbrone Jr MA, Loskutoff DJ. Regulation of the fibrinolytic system of cultured human vascular endothelium by interleukin 1. J Clin Invest 1986; 78: 587-591
• 38 De Groot PG, Verweij CL, Nawroth PP, de Boer HC, Stem DM, Sixma JJ. Interleukin-1 inhibits the synthesis of von Willebrand factor in endothelial cells, which results in a decreased reactivity of their matrix toward platelets. Arteriosclerosis 1987; 7: 605-611
• 39 Giddings JC, Shall L. Enhanced release of von Willebrand factor by human endothelial cells in culture in the presence of phorbol myristate acetate and interleukin 1. Thromb Res 1987; 47: 259-267
• 40 Schorer AE, Moldow CF, Rick ME. Interleukin 1 or endotoxin increases the release of von Willebrand factor from human endothelial cells. Br J Haematol 1987; 67: 193-197