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Thromb Haemost 2006; 96(05): 665-670
DOI: 10.1160/TH06-06-0321
Cardiovascular Biology and Cell Signalling
Schattauer GmbH

Plasminogen activator inhibitor-1 impairs plasminogen activationmediated vascular smooth muscle cell apoptosis

Patrick Rossignol
1   Center for Molecular and Vascular Biology, K.U. Leuven, Leuven, Belgium
2   INSERM, U765, Paris, France
3   AP-HP, Service de Médecine Vasculaire et Hypertension Artérielle, Hôpital Européen Georges Pompidou, Paris, France
4   Université Paris Descartes, Faculté de Médecine René Descartes, Paris, France
,
Eduardo Anglès-Cano
5   INSERM, U698, Paris, France
6   Université Paris7-Denis Diderot, Faculté de médecine, Site Xavier Bichat, IFR2, CIC7, and Université Paris 13, Paris, France
,
Henri Roger Lijnen
1   Center for Molecular and Vascular Biology, K.U. Leuven, Leuven, Belgium
› Author AffiliationsFinancial support: Patrick Rossignol received a grant from the Institut National de la Santé et de la Recherche Médicale (INSERM, France). This study was supported by the Leducq Foundation (Transatlantic Networks).
Further Information

Publication History

Received 12 June 2006

Accepted after resubmission 14 September 2006

Publication Date:
01 December 2017 (online)

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Summary

The role of plasminogen activator inhibitor-1 (PAI-1) in vascular smooth muscle cell (VSMC) apoptosis mediated by plasminogen activation was studied with the use of aorticVSMC derived from mice with deficiency of PAI-1 (PAI-1-/-), tissue-type (t-PA-/-) or urokinase-type (u-PA-/-) plasminogen activator or from wildtype (WT) mice with corresponding genetic background. Plasminogen incubated with confluentVSMC was activated ina concentration-dependent and saturable manner for all four cell types, with maximal activation rates that were comparable for WT,u-PA-/and t-PA-/cells,but about two-fold higher for PAI-1-/cells. Plasminogen activation was impaired by addition of the lysine analogue 6-aminohexanoic acid, and by addition of t-PA and u-PA neutralizing antibodies, suggesting that it depends on binding to cell surface COOH-terminal lysine residues, and on plasminogen activator activity. Morphological alterations consistent with apoptosis were observed much earlier in PAI-1-/than in WT VSMC. Without addition of plasminogen, the apoptotic index was similar for all four cell types, whereas after incubation with physiological plasminogen concentrations, it was greater in PAI-1-/VSMC, as compared to WT, t-PA-/or u-PA-/VSMC. Furthermore, the apoptotic rate paralleled the release of plasmin. Thus, plasmin-mediated apoptosis of VSMC occurs via plasminogen activation by either t-PA or u-PA and is impaired by PAI-1.

Keywords

Apoptosis - proteolysis / pericellular - plasminogen activators - plasminogen activator inhibitors - plasminogen (-fragments)
 

  • References

  • 1 Geng YJ, Libby P. Evidence for apoptosis in advanced human atheroma. Colocalization with interleukin-1 beta-converting enzyme. AmJ Pathol 1995; 147: 251-66.
    PubMedSearch in Google Scholar
  • 2 Han DK, Haudenschild CC, Hong MK. et al. Evidence for apoptosis in human atherogenesis and in a rat vascular injury model. Am J Pathol 1995; 147: 267-77.
    PubMedSearch in Google Scholar
  • 3 Bauriedel G, Hutter R, Welsch U. et al. Role of smooth muscle cell death in advanced coronary primary lesions: implications for plaque instability. Cardiovasc Res 1999; 41: 480-8.
    CrossrefPubMedSearch in Google Scholar
  • 4 Michel JB. Anoïkis in the cardiovascular system. Known and unknown extracellular mediators. Arterioscler Thromb Vasc Biol 2003; 23: 2146-54.
    CrossrefPubMedSearch in Google Scholar
  • 5 Houard X, Monnot C, Dive V. et al. Vascuar smooth muscle cells efficiently activate a new proteinase cascade involving plasminogen and fibronectin. J Cell Biochem 2003; 88: 1188-201.
    CrossrefPubMedSearch in Google Scholar
  • 6 Davis J, Wagner MR, Zhang W. et al. Amyloid betaprotein stimulates the expression of urokinase-type plasminogen activator (uPA) and its receptor (uPAR) in human cerebrovascular smooth muscle cells. J Biol Chem 2003; 278: 19054-61.
    CrossrefPubMedSearch in Google Scholar
  • 7 Meilhac O, Ho-Tin-Noe B, Houard X. et al. Pericellular plasmin induces smooth muscle cell anoikis. Faseb J 2003; 17: 1301-3.
    CrossrefPubMedSearch in Google Scholar
  • 8 Lijnen HR. Plasmin and matrix metalloproteinases in vascular remodeling. Thromb Haemost 2001; 86: 324-33.
    Thieme ConnectPubMedSearch in Google Scholar
  • 9 Grainger DJ, Kemp PR, Liu AC. et al. Activation of transforming growth factor-beta is inhibited in transgenic apolipoprotein(a) mice. Nature 1994; 370: 460-2.
    CrossrefPubMedSearch in Google Scholar
  • 10 Bonnefoy A, Legrand C. Proteolysis of subendothelial adhesive glycoproteins (fibronectin, thrombospondin, and von Willebrand factor) by plasmin, leukocyte cathepsin G, and elastase. Thromb Res 2000; 98: 323-32.
    CrossrefPubMedSearch in Google Scholar
  • 11 Liotta LA, Goldfarb RH, Terranova VP. Cleavage of laminin by thrombin and plasmin: alpha thrombin selectively cleaves the beta chain of laminin. Thromb Res 1981; 21: 663-73.
    CrossrefPubMedSearch in Google Scholar
  • 12 Rossignol P, Luttun A, Martin-Ventura JL. et al. Plasminogen activation: a mediator of vascular smooth muscle cell apoptosis in atherosclerotic plaques. J Thromb Haemost 2006; 04: 664-70.
    CrossrefPubMedSearch in Google Scholar
  • 13 Carmeliet P, Kieckens L, Schoonjans L. et al. Plasminogen activator inhibitor-1 gene-deficient mice. I. Generation by homologous recombination and characterization. J Clin Invest 1993; 92: 2746-55.
    CrossrefPubMedSearch in Google Scholar
  • 14 Carmeliet P, Schoonjans L, Kieckens L. et al. Physiological consequences of loss of plasminogen activator gene function in mice. Nature 1994; 368: 419-24.
    CrossrefPubMedSearch in Google Scholar
  • 15 Ploplis VA, Carmeliet P, Vazirzadeh S. et al. Effects of disruption of the plasminogen gene on thrombosis, growth, and health in mice. Circulation 1995; 92: 2585-93.
    CrossrefPubMedSearch in Google Scholar
  • 16 Giles AR. Guidelines for the use of animals in biomedical research. Thromb Haemost 1987; 58: 1078-84.
    Thieme ConnectPubMedSearch in Google Scholar
  • 17 Lijnen HR, Van Hoef B, Beelen V. et al. Characterization of the murine plasma fibrinolytic system. Eur J Biochem 1994; 224: 863-71.
    CrossrefPubMedSearch in Google Scholar
  • 18 Lijnen HR, Van Hoef B, Collen D. Characterization of the murine plasminogen/urokinase-type plasminogen activator system. Eur J Biochem 1996; 241: 840-8.
    CrossrefPubMedSearch in Google Scholar
  • 19 Declerck PJ, Verstreken M, Collen D. Immunoassay of murine t-PA, u-PA and PAI-1 using monoclonal antibodies raised in gene-inactivated mice. Thromb Haemost 1995; 74: 1305-9.
    Thieme ConnectPubMedSearch in Google Scholar
  • 20 Lijnen HR, Silence J, Lemmens G. et al. Regulation of gelatinase activity in mice with targeted inactivation of components of the plasminogen/plasmin system. Thromb Haemost 1998; 79: 1171-6.
    Thieme ConnectPubMedSearch in Google Scholar
  • 21 Heussen C, Dowdle EB. Electrophoretic analysis of plasminogen activators in polyacrylamide gels containing sodium dodecyl sulfate and copolymerized substrates. Anal Biochem 1980; 102: 196-202.
    CrossrefPubMedSearch in Google Scholar
  • 22 Lijnen HR, Okada K, Matsuo O. et al. Alpha2-antiplasmin gene deficiency in mice is associated with enhanced fibrinolytic potential without overt bleeding. Blood 1999; 93: 2274-81.
    PubMedSearch in Google Scholar
  • 23 Denizot F, Lang R. Rapid colorimetric assay for cell growth and survival. Modifications to the tetrazolium dye procedure giving improved sensitivity and reliability.J Immunol Methods 1986; 89: 271-7.
    PubMedSearch in Google Scholar
  • 24 Adderley SR, Fitzgerald DJ. Glycoprotein IIb/IIIa antagonists induce apoptosis in rat cardiomyocytes by caspase-3 activation. J Biol Chem 2000; 275: 5760-6.
    CrossrefPubMedSearch in Google Scholar
  • 25 Al-Fakhri N, Chavakis T, Schmidt-Woll T. et al. Induction of apoptosis in vascular cells by plasminogen activator inhibitor-1 and high molecular weight kininogen correlates with their anti-adhesive properties. Biol Chem 2003; 384: 423-35.
    PubMedSearch in Google Scholar
  • 26 Kwaan HC, Wang J, Svoboda K. et al. Plasminogen activator inhibitor 1 may promote tumour growth through inhibition of apoptosis. Br J Cancer 2000; 82: 1702-8.
    CrossrefPubMedSearch in Google Scholar
  • 27 Davis GE, Pintar KAAllen, Salazar R. et al. Matrix metalloproteinase-1 and -9 activation by plasmin regulates a novel endothelial cell-mediated mechanism of collagen gel contraction and capillary tube regression in threedimensional collagen matrices. J Cell Sci 2001; 114: 917-30.
    PubMedSearch in Google Scholar
  • 28 Rossignol P, Ho-Tin-Noe B, Vranckx R. et al. Protease-nexin-1 inhibits plasminogen activation-induced apoptosis of adherent cells. J Biol Chem 2004; 279: 10346-56.
    CrossrefPubMedSearch in Google Scholar
  • 29 Zhang X, Chaudhry A, Chintala SK. Inhibition of plasminogen activation protects against ganglion cells loss in a mouse model of retinal damage. Mol Vis 2003; 09: 238-48.
    PubMedSearch in Google Scholar
  • 30 Reijerkerk A, Mosnier LO, Kranenburg O. et al. Amyloid endostatin induces endothelial cell detachment by stimulation of the plasminogen activation system. Mol Cancer Res 2003; 01: 561-8.
    PubMedSearch in Google Scholar
  • 31 Hishikawa K, Nakaki T, Fujii T. Transforming growth factor-beta induces apoptosis via connective tissue growth factor in human aortic smooth muscle cells. Eur J Pharmacol 1999; 385: 287-90.
    CrossrefPubMedSearch in Google Scholar