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
› InstitutsangabenFinancial 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).
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.
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