Download PDF
Thromb Haemost 2001; 85(05): 915-923
DOI: 10.1055/s-0037-1615768
Review Article
Schattauer GmbH

Human Endothelial Cells Maintain Anti-Aggregatory Activity for Platelets during Apoptosis

Wei Xu
1   Department of Oral Pathology and Oral Medicine, University of Sydney, Westmead Hospital Dental Clinical School, Westmead Hospital, NSW, Australia
,
Emmanuel J. Favaloro
2   Department of Haematology, Westmead Hospital, Australia
,
Heather Medbury
3   Department of Surgery, Westmead Hospital, Australia
,
Hans Zoellner
1   Department of Oral Pathology and Oral Medicine, University of Sydney, Westmead Hospital Dental Clinical School, Westmead Hospital, NSW, Australia
› Author Affiliations
Further Information

Publication History

Received 14 August 2001

Accepted after revision 05 December 2000

Publication Date:
11 December 2017 (online)

Also available at
    Permissions and Reprints

Summary

Despite evidence of elevated levels of tissue factor and platelet binding by apoptotic endothelial cells, microthrombi do not appear to be associated with apoptotic endothelium and this suggests maintained anti-aggregatory activity for platelets. We report that anti-aggregatory activity is maintained by apoptotic endothelium obtained by serum and or matrix deprivation, which we propose as models for apoptotic endothelial cells released during microvascular remodelling and traumatic detachment respectively. Both apoptotic and non-apoptotic endothelium had strong anti-aggregatory activity for platelets stimulated with either ADP or thrombin. Inhibition experiments using L-NAME and indo-methacin indicated a role for nitric oxide and prostacyclin in this activity. Experiments with latex beads further confirmed that inhibited platelet aggregation by endothelium was not merely a non-specific phenomenon. These data support the idea that EC maintain active anti-thrombotic activity during apoptosis, consistent with maintained urokinase levels and canalicular fragmentation reported elsewhere.

Keywords

Endothelium - apoptosis - platelets - aggregation
 

  • References

  • 1 Hubbard AR, Jennings CA, Barrowcliffe TW. Anticoagulant properties in vitro of heparan sulphates. Thromb Res 1984; 35: 567-76.
    CrossrefPubMedSearch in Google Scholar
  • 2 Hawiger J. Formation and regulation of platelet and fibrin hemostatic plug. Hum Pathol 1987; 18: 111-22.
    CrossrefPubMedSearch in Google Scholar
  • 3 Dittman WA, Majerus PW. Structure and function of thrombomodulin: a natural anticoagulant. Blood 1990; 75: 329-36.
    PubMedSearch in Google Scholar
  • 4 Smith EB. Haemostatic factors and atherogenesis. Atherosclerosis 1996; 124: 137-43.
    CrossrefPubMedSearch in Google Scholar
  • 5 Zoellner H, Hofler M, Beckmann R, Bielek E, Vanyek E, Kumabashiri I, Binder B. Fibrinolytic proteins in apoptotic human umbilical vein endothelial cells. Thromb Res 1998; 91: 209-19.
    CrossrefPubMedSearch in Google Scholar
  • 6 Hoak JC, Parks WM, Fry GL, Brotherton AA, Czervionke RL. Role of the vascular endothelium. Progr Clin Biol Res 1982; 89: 281-94.
    PubMedSearch in Google Scholar
  • 7 Radomski MW, Palmer RM, Moncada S. Comparative pharmacology of endothelium-derived relaxing factor, nitric oxide and prostacyclin in platelets. Brit J Pharmacol 1987; 92: 181-7.
    CrossrefPubMedSearch in Google Scholar
  • 8 Schiffrin EL. The endothelium and control of blood vessel function in health and disease. Clin Invest Med 1994; 17: 602-20.
    PubMedSearch in Google Scholar
  • 9 Heinmoller E, Weinel RJ, Heidtmann HH, Salge U, Seitz R, Schmitz I, Muller KM, Zirngibl H. Studies on tumor-cell-induced platelet aggregation in human lung cancer cell lines. J Cancer Res Clin Oncol 1996; 122: 735-44.
    CrossrefPubMedSearch in Google Scholar
  • 10 Bevilacqua MP, Pober JS, Majeau GR, Cotran RS, Gimbrone MJ. Interleukin 1 (IL-1) induces biosynthesis and cell surface expression of procoagulant activity in human vascular endothelial cells. J Exp Med 1984; 160: 618-23.
    CrossrefPubMedSearch in Google Scholar
  • 11 Bevilacqua MP, Pober JS, Majeau GR, Fiers W, Cotran RS, Gimbrone MJ. Recombinant tumor necrosis factor induces procoagulant activity in cultured human vascular endothelium: characterization and comparison with the actions of interleukin 1. Proc Nat Acad Sci USA 1985; 83: 4533-7.
    PubMedSearch in Google Scholar
  • 12 Nawroth PP, Stern DM. Modulation of endothelial cell hemostatic properties by tumor necrosis factor. J Exp Med 1986; 163: 740-5.
    CrossrefPubMedSearch in Google Scholar
  • 13 Colucci M, Balconi G, Lorenzet R, Pietra A, Locati D, Donati MB, Semeraro N. Cultured human endothelial cells generate tissue factor in response to endotoxin. J Clin Invest 1983; 71: 1893-6.
    CrossrefPubMedSearch in Google Scholar
  • 14 Moore KL, Andreoli SP, Esmon NL, Esmon CT, Bang NU. Endotoxin enhances tissue factor and suppresses thrombomodulin expression of human vascular endothelium in vitro. J Clin Invest 1987; 79: 124-30.
    CrossrefPubMedSearch in Google Scholar
  • 15 Walker NI, Gobe GC. Cell death and cell proliferation during atrophy of the rat parotid gland induced by duct obstruction. J Pathol 1987; 153: 333-44.
    CrossrefPubMedSearch in Google Scholar
  • 16 Walker NI, Bennett RE, Kerr JF. Cell death by apoptosis during involution of the lactating breast in mice and rats. Am J Anat 1989; 185: 19-32.
    CrossrefPubMedSearch in Google Scholar
  • 17 Sgonc R, Gruschwitz MS, Dietrich H, Recheis H, Gershwin ME, Wick G. Endothelial cell apoptosis is a primary pathogenetic event underlying skin lesions in avian and human scleroderma. J Clin Invest 1996; 98: 785-92.
    CrossrefPubMedSearch in Google Scholar
  • 18 Meeson A, Palmer M, Calfon M, Lang R. A relationship between apoptosis and flow during programmed capillary regression is revealed by vital analysis. Development 1996; 122: 3929-38.
    PubMedSearch in Google Scholar
  • 19 Darby IA, Bisucci T, Hewitson TD, MacLellan DG. Apoptosis is increased in a model of diabetes-impaired wound healing in genetically diabetic mice. Int J Biochem Cell Biol 1997; 29: 191-200.
    CrossrefPubMedSearch in Google Scholar
  • 20 Tatarczuch L, Philip C, Lee CS. Involution of the sheep mammary gland. J Anat 1997; 190: 405-16.
    CrossrefPubMedSearch in Google Scholar
  • 21 Dimmeler S, Hermann C, Zeiher AM. Apoptosis of endothelial cells. Contribution to the pathophysiology of atherosclerosis? Eur Cytokine Network 1998; 9: 697-8.
    PubMedSearch in Google Scholar
  • 22 Gobe G, Browning J, Howard T, Hogg N, Winterford C, Cross R. Apoptosis occurs in endothelial cells during hypertension-induced microvascular rarefaction. J Struct Biol 1997; 118: 63-72.
    CrossrefPubMedSearch in Google Scholar
  • 23 Lai X, Liu Y, Wang J, Li S, Chen L, Guan Z. Injury to vascular endothelial cells and the change of plasma endothelin level in dogs with gunshot wounds. J Trauma-Injury Infec Crit Care 1996; 40: S60-S62.
    PubMedSearch in Google Scholar
  • 24 Meredith JJ, Fazeli B, Schwartz MA. The extracellular matrix as a cell survival factor. Mol Biol Cell 1993; 4: 953-61.
    CrossrefPubMedSearch in Google Scholar
  • 25 Zoellner H, Hofler M, Beckmann R, Hufnagl P, Vanyek E, Bielek E, Wojta J, Fabry A, Lockie S, Binder BR. Serum albumin is a specific inhibitor of apoptosis in human endothelial cells. J Cell Sci 1996; 109: 2571-80.
    PubMedSearch in Google Scholar
  • 26 Solovey A, Gui L, Ramakrishnan S, Steinberg MH, Hebbel RP. Sickle cell anemia as a possible state of enhanced anti-apoptotic tone: survival effect of vascular endothelial growth factor on circulating and unanchored endothelial cells. Blood 1999; 93: 3824-30.
    PubMedSearch in Google Scholar
  • 27 Araki S, Simada Y, Kaji K, Hayashi H. Role of protein kinase C in the inhibition by fibroblast growth factor of apoptosis in serum-depleted endothelial cells. Biochem Biophys Res Comm 1990; 172: 1081-5.
    CrossrefPubMedSearch in Google Scholar
  • 28 Zoellner H, Hou JY, Lovery M, Kingham J, Srivastava M, Bielek E, Vanyek E, Binder BR. Inhibition of microvascular endothelial apoptosis in tissue explants by serum albumin. Microvasc Res 1999; 57: 162-73.
    CrossrefPubMedSearch in Google Scholar
  • 29 Araki S, Shimada Y, Kaji K, Hayashi H. Apoptosis of vascular endothelial cells by fibroblast growth factor deprivation. Biochem Biophys Res Commun 1990; 168: 1194-200.
    CrossrefPubMedSearch in Google Scholar
  • 30 Sandison JC. Observations on the growth of blood vessels as seen in the transparent chamber introduced into the rabbit’s ear. Am J Anat 1928; 41: 475-96.
    CrossrefPubMedSearch in Google Scholar
  • 31 Kerr JF, Wyllie AH, Currie AR. Apoptosis: a basic biological phenomenon with wide-ranging implications in tissue kinetics. Brit J Cancer 1972; 26: 239-57.
    CrossrefPubMedSearch in Google Scholar
  • 32 Vaux DL, Strasser A. The molecular biology of apoptosis. Proc Natl Acad Sci USA 1996; 19 (Suppl. 93) 2239-44.
    PubMedSearch in Google Scholar
  • 33 Poirier J. Apoptosis techniques and protocol. Totowa, NJ: Humana Press; 1997
    Search in Google Scholar
  • 34 Martins L, Earnshaw WC. Apoptosis: alive and kicking in 1997. Trends in Cell Biol 1997; 7: 111-4.
    CrossrefPubMedSearch in Google Scholar
  • 35 Wyllie AH. Glucocorticoid-induced thymocyte apoptosis is associated with endogenous endonuclease activation. Nature 1980; 284: 555-6.
    CrossrefPubMedSearch in Google Scholar
  • 36 Wyllie AH, Morris RG, Smith AL, Dunlop D. Chromatin cleavage in apoptosis: association with condensed chromatin morphology and dependence on macromolecular synthesis. J Pathol 1984; 142: 67-77.
    CrossrefPubMedSearch in Google Scholar
  • 37 Arends MJ, Morris RG, Wyllie AH. Apoptosis. The role of the endonuclease. Am J Pathol 1990; 136: 593-608.
    PubMedSearch in Google Scholar
  • 38 Zoellner H, Bielek E, Vanyek E, Fabry A, Wojta J, Hofler M, Binder B. Canalicular fragmentation of apoptotic human endothelial cells. Endothelium 1996; 4: 177-88.
    CrossrefPubMedSearch in Google Scholar
  • 39 Greeno EW, Bach RR, Moldow CF. Apoptosis is associated with increased cell surface tissue factor procoagulant activity. Lab Invest 1996; 75: 281-9.
    PubMedSearch in Google Scholar
  • 40 Bombeli T, Karsan A, Tait JF, Harlan JM. Apoptotic vascular endothelial cells become procoagulant. Blood 1997; 89: 2429-42.
    PubMedSearch in Google Scholar
  • 41 Casciola-Rosen L, Rosen A, Petri M, Schlissel M. Surface blebs on apoptotic cells are sites of enhanced procoagulant activity: implications for coagulation events and antigenic spread in systemic lupus erythematosus. Proc Natl Acad Sci USA 1996; 93: 1624-9.
    CrossrefPubMedSearch in Google Scholar
  • 42 de GG, Cerletti C, Dejana E, Vermylen J. Current issues in thrombosis prevention with antiplatelet drugs. Drugs 1986; 31: 517-49.
    CrossrefPubMedSearch in Google Scholar
  • 43 Ridker PM, Willich SN, Muller JE, Hennekens CH. Aspirin, platelet aggregation, and the circadian variation of acute thrombotic events. Chronobiology Int 1991; 8: 327-35.
    CrossrefPubMedSearch in Google Scholar
  • 44 Bombeli T, Schwartz BR, Harlan JM. Endothelial cells undergoing apoptosis become proadhesive for nonactivated platelets. Blood 1999; 93: 3831-8.
    PubMedSearch in Google Scholar
  • 45 Maciag T, Hoover GA, Stemerman MB, Weinstein R. Factors which stimulate the growth of human umbilical vein endothelial cells in vitro. In: Jaffe EA. ed. Biology of Endothelial cells. Nijhoff, M.; Dordrecht: 1984: 87-96.
    Search in Google Scholar
  • 46 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: 2745-56.
    CrossrefPubMedSearch in Google Scholar
  • 47 Zoellner H, Wojta J, Gallicchio M, McGrath K, Hamilton JA. Cytokine regulation of the synthesis of plasminogen activator inhibitor-2 by human vascular endothelial cells. Comparison with plasminogen activator inhibitor-1 synthesis. Thromb Haemost 1993; 69: 135-40.
    Thieme ConnectPubMedSearch in Google Scholar
  • 48 Jaffe EA. Culture of human endothelial cells. Transplantation Proc 1980; 12: 49-53.
    PubMedSearch in Google Scholar
  • 49 Holthofer H, Virtanen I, Kariniemi AL, Hormia M, Linder E, Miettinen A. Ulex europaeus I lectin as a marker for vascular endothelium in human tissues. Lab Invest 1982; 47: 60-6.
    PubMedSearch in Google Scholar
  • 50 Rasko JEJ, North K, Favaloro EJ, Grispo L, Berndt MC. Attenuated platelet sensitivity to collagen in patients with Neurofibromatosis Type 1. Br J Haematol 1995; 89: 582-8.
    CrossrefPubMedSearch in Google Scholar
  • 51 Wilson A, Leeder S, Koutts J, Heller R, Exner T, Dinale A. Platelet aggregation and coronary heart disease risk factor variation in Australian populations with different coronary heart disease mortality. Annal Epidemiol 1992; 495-508.
    PubMedSearch in Google Scholar
  • 52 Rees DD, Palmer RM, Schulz R, Hodson HF, Moncada S. Characterization of three inhibitors of endothelial nitric oxide synthase in vitro and in vivo. Brit J Pharmacol 1990; 101: 746-52.
    CrossrefPubMedSearch in Google Scholar
  • 53 Laneuville O, Breuer DK, Dewitt DL, Hla T, Funk CD, Smith WL. Differential inhibition of human prostaglandin endoperoxide H synthases-1 and -2 by nonsteroidal anti-inflammatory drugs. J Pharmacol Exp Ther 1994; 271: 927-34.
    PubMedSearch in Google Scholar
  • 54 Born G, Cross M. The aggregation of blood platelets. J Physiol 1963; 178-95.
    PubMedSearch in Google Scholar
  • 55 Cejna M, Fritsch G, Printz D, Schulte-Hermann R, Bursch W. Kinetics of apoptosis and secondary necrosis in cultured rat thymocytes and S. 49 mouse lymphoma and CEM human leukemia cells. Biochem Cell Biol 1994; 72: 677-85.
    CrossrefPubMedSearch in Google Scholar
  • 56 Hebert MJ, Takano T, Holthofer H, Brady HR. Sequential morphologic events during apoptosis of human neutrophils. Modulation by lipoxygenase-derived eicosanoids. J Immun 1996; 157: 3105-15.
    PubMedSearch in Google Scholar
  • 57 Papassotiropoulos A, Ludwig M, Naib-Majani W, Rao GS. Induction of apoptosis and secondary necrosis in rat dorsal root ganglion cell cultures by oxidized low density lipoprotein. Neurosci Lett 1996; 209: 33-6.
    CrossrefPubMedSearch in Google Scholar
  • 58 Shah KA, Shurey S, Green CJ. Apoptosis after intestinal ischemia-reper-fusion injury: a morphological study. Transplantation 1997; 64: 1393-7.
    CrossrefPubMedSearch in Google Scholar
  • 59 Harker LA, Hanson SR, Runge MS. Thrombin hypothesis of thrombus generation and vascular lesion formation. Am J Cardiol 1995; 75: 12-7.
    CrossrefPubMedSearch in Google Scholar
  • 60 Puri RN, Colman RW. ADP-induced platelet activation. Crit Rev Biochem Mol Biol 1997; 32: 437-502.
    CrossrefPubMedSearch in Google Scholar
  • 61 Bankowski E, Niewiarowski S, Galasinski W. Platelet aggregation by human collagen in relation to its age. Gerontologia 1967; 13: 219-26.
    CrossrefPubMedSearch in Google Scholar
  • 62 Madri JA, Dreyer B, Pitlick FA, Furthmayr H. The collagenous components of the subendothelium. Correlation of structure and function. Lab Invest 1980; 43: 303-15.
    PubMedSearch in Google Scholar
  • 63 Trelstad RL. Special state of the fibril end: site of growth, point of cell surface attachment, and possible site for platelet interaction. In: Gastpar H. ed. Collagen-Platelet Interaction. New York: Schattauer Verlag; 1978: 153-60.
    Search in Google Scholar
  • 64 Radomski MW, Palmer RM, Moncada S. The role of nitric oxide and cGMP in platelet adhesion to vascular endothelium. Biochem Biophys Res Comm 1987; 148: 1482-9.
    CrossrefPubMedSearch in Google Scholar
  • 65 Sneddon JM, Vane JR. Endothelium-derived relaxing factor reduces platelet adhesion to bovine endothelial cells. Proc Natl Acad Sci USA 1988; 85: 2800-4.
    CrossrefPubMedSearch in Google Scholar
  • 66 Palmer RM, Ashton DS, Moncada S. Vascular endothelial cells synthesize nitric oxide from L-arginine. Nature 1988; 333: 664-6.
    CrossrefPubMedSearch in Google Scholar
  • 67 Cote YP, Filep JG, Battistini B, Gauvreau J, Sirois P, Beaudoin AR. Characterization of ATP-diphosphohydrolase activities in the intima and media of the bovine aorta: evidence for a regulatory role in platelet activation in vitro. Biochim Biophys Acta 1992; 1139: 133-42.
    CrossrefPubMedSearch in Google Scholar
  • 68 Marcus AJ, Broekman MJ, Drosopoulos JH, Islam N, Alyonycheva TN, Safier LB, Hajjar KA, Posnett DN, Schoenborn MA, Schooley KA, Gayle RB, Maliszewski CR. The endothelial cell ecto-ADPase responsible for inhibition of platelet function is CD39. J Clin Invest 1997; 99: 1351-60.
    CrossrefPubMedSearch in Google Scholar
  • 69 Robson SC, Kaczmarek E, Siegel JB, Candinas D, Koziak K, Millan M, Hancock WW, Bach FH. Loss of ATP diphosphohydrolase activity with endothelial cell activation. J Exp Med 1997; 185: 153-63.
    CrossrefPubMedSearch in Google Scholar
  • 70 Yagi K, Shinbo M, Hashizume M, Shimba LS, Kurimura S, Miura Y. ATP diphosphohydrolase is responsible for ecto-ATPase and ecto-ADPase activities in bovine aorta endothelial and smooth muscle cells. Biochem Biophys Res Comm 1991; 180: 1200-6.
    CrossrefPubMedSearch in Google Scholar