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

Patrick Rossignol; Eduardo Anglès-Cano; Henri Roger Lijnen

doi:10.1160/TH06-06-0321

Thrombosis and Haemostasis, Table of Contents

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

¹Center for Molecular and Vascular Biology, K.U. Leuven, Leuven, Belgium

²INSERM, U765, Paris, France

³AP-HP, Service de Médecine Vasculaire et Hypertension Artérielle, Hôpital Européen Georges Pompidou, Paris, France

⁴Université Paris Descartes, Faculté de Médecine René Descartes, Paris, France

,

Eduardo Anglès-Cano

⁵INSERM, U698, Paris, France

⁶Université Paris7-Denis Diderot, Faculté de médecine, Site Xavier Bichat, IFR2, CIC7, and Université Paris 13, Paris, France

,

Henri Roger Lijnen

¹Center for Molecular and Vascular Biology, K.U. Leuven, Leuven, Belgium

› Author Affiliations

Abstract

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)

Full Text

References

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.
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.
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.
4 Michel JB. Anoïkis in the cardiovascular system. Known and unknown extracellular mediators. Arterioscler Thromb Vasc Biol 2003; 23: 2146-54.
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.
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.
7 Meilhac O, Ho-Tin-Noe B, Houard X. et al. Pericellular plasmin induces smooth muscle cell anoikis. Faseb J 2003; 17: 1301-3.
8 Lijnen HR. Plasmin and matrix metalloproteinases in vascular remodeling. Thromb Haemost 2001; 86: 324-33.
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.
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.
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.
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.
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.
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.
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.
16 Giles AR. Guidelines for the use of animals in biomedical research. Thromb Haemost 1987; 58: 1078-84.
17 Lijnen HR, Van Hoef B, Beelen V. et al. Characterization of the murine plasma fibrinolytic system. Eur J Biochem 1994; 224: 863-71.
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.
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.
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.
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.
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.
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.
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.
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.
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.
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.
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.
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.
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.
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.