Glycoprotein IIb/IIIa antagonists induce apoptosis in rat cardiomyocytes by caspase-3 activation

被引:83
作者
Adderley, SR [1 ]
Fitzgerald, DJ [1 ]
机构
[1] Royal Coll Surgeons Ireland, Ctr Cardiovasc Sci, Dublin 2, Ireland
关键词
D O I
10.1074/jbc.275.8.5760
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
The platelet integrin glycoprotein (GP) IIb/IIIa, which mediates platelet aggregation, has been the target for novel antiplatelet agents, the GPIIb/IIIa antagonists. Several GPIIb/IIIa antagonists have been developed based on the peptide RGDS present in adhesion proteins, including the principle ligand fibrinogen. The apoptosis enzyme, procaspase-3, contains an RGD-recognition sequence and is activated by RGDS. We examined the effects of RGDS and several GPIIb/IIIa antagonists on cell death and procaspase-3 activation in rat neonatal cardiomyocytes, These antagonists do not recognize rat integrins, yet RGDS, orbofiban, and xemilofiban induced dose-dependent apoptosis and procaspase-3 activation in cardiomyocytes over 72 h, particularly under hypoxic conditions. Scrambled peptide, the monoclonal antibody 7E3 or integrelin (a peptide containing a KGD sequence), had little or no effect. Immunoprecipitation of procaspase-3 followed by treatment with the compounds showed that procaspase-3 was activated directly by RGDS, orbofiban, xemilofiban, and by monoclonal 7E3 antibody, the latter demonstrating that compounds must enter cells to induce apoptosis through caspase activation. Integrelin had no effect. Binding studies with H-3-SC52012B, a GPIIb/IIIa antagonist analogue of orbofiban, showed no specific binding to cardiomyocytes, but the radioligand accumulated intracellularly over 72 h. H-3-SC52012B also bound directly to human recombinant caspase-3 (K-d, 59 +/- 2 nm), and this was prevented by orbofiban, xemilofiban, and the monoclonal 7E3 antibody but not by integrelin. Finally confocal microscopy showed that RGDS co-localized with caspase-3 inside the cell. These data show that RGDS and its mimetics induce cardiomyocyte apoptosis by direct activation of procaspase-3.
引用
收藏
页码:5760 / 5766
页数:7
相关论文
共 30 条
[1]   Hypoxia induces apoptosis in human neuroblastoma SK-N-MC cells by caspase activation accompanying cytochrome c release from mitochondria [J].
Araya, R ;
Uehara, T ;
Nomura, Y .
FEBS LETTERS, 1998, 439 (1-2) :168-172
[2]  
BOUKERCHE H, 1989, BLOOD, V74, P909
[3]  
BRADFORD MM, 1976, ANAL BIOCHEM, V72, P248, DOI 10.1016/0003-2697(76)90527-3
[4]   INTEGRIN ALPHA(V)BETA(3) ANTAGONISTS PROMOTE TUMOR-REGRESSION BY INDUCING APOPTOSIS OF ANGIOGENIC BLOOD-VESSELS [J].
BROOKS, PC ;
MONTGOMERY, AMP ;
ROSENFELD, M ;
REISFELD, RA ;
HU, TH ;
KLIER, G ;
CHERESH, DA .
CELL, 1994, 79 (07) :1157-1164
[5]   RGD peptides induce apoptosis by direct caspase-3 activation [J].
Buckley, CD ;
Pilling, D ;
Henriquez, NV ;
Parsonage, G ;
Threlfall, K ;
Scheel-Toellner, D ;
Simmons, DL ;
Albar, AN ;
Lord, JM ;
Salmon, M .
NATURE, 1999, 397 (6719) :534-539
[6]   ABOLITION OF INVIVO PLATELET THROMBUS FORMATION IN PRIMATES WITH MONOCLONAL-ANTIBODIES TO THE PLATELET GPIIB-IIIA RECEPTOR - CORRELATION WITH BLEEDING-TIME, PLATELET-AGGREGATION, AND BLOCKADE OF GPIIB-IIIA RECEPTORS [J].
COLLER, BS ;
FOLTS, JD ;
SMITH, SR ;
SCUDDER, LE ;
JORDAN, R .
CIRCULATION, 1989, 80 (06) :1766-1774
[7]   THE PHARMACOLOGY OF THE INTEGRINS [J].
COX, D ;
AOKI, T ;
SEKI, J ;
MOTOYAMA, Y ;
YOSHIDA, K .
MEDICINAL RESEARCH REVIEWS, 1994, 14 (02) :195-228
[8]  
Cox D, 1999, THROMB HAEMOSTASIS, P727
[9]  
DU XP, 1995, NATURE, V376, P37
[10]   2 INTEGRIN-BINDING PEPTIDES ABROGATE T-CELL-MEDIATED IMMUNE-RESPONSES INVIVO [J].
FERGUSON, TA ;
MIZUTANI, H ;
KUPPER, TS .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1991, 88 (18) :8072-8076