Morphological and Molecular Characterization of Adult Cardiomyocyte Apoptosis During Hypoxia and Reoxygenation

• Published in: Circulation research 2000-07, Vol.87 (2), p.118-125
• Main Authors: Kang, Peter M, Haunstetter, Armin, Aoki, Hiroki, Usheva, Anny, Izumo, Seigo
• Format: Article
• Language: English
• Subjects: Aerobiosis; Amino Acid Chloromethyl Ketones - pharmacology; Animals; Apoptosis - physiology; Biological and medical sciences; Cardiology. Vascular system; Caspase Inhibitors; Cell Hypoxia - physiology; Coronary heart disease; Cytochrome c Group - metabolism; Female; Genes, bcl-2; Heart; Heart - physiology; Humans; Medical sciences; Mitochondria, Heart - metabolism; Mitochondria, Heart - ultrastructure; Myocardium - cytology; Myocardium - metabolism; Myocardium - ultrastructure; Proto-Oncogene Proteins c-bcl-2 - genetics; Proto-Oncogene Proteins c-bcl-2 - metabolism; Rats; Rats, Sprague-Dawley; Recombinant Proteins - metabolism; Transfection
• ISSN: 0009-7330; 1524-4571
• DOI: 10.1161/01.res.87.2.118

Apoptosis has been implicated in ischemic heart disease, but its mechanism in cardiomyocytes has not been elucidated. In this study, we investigate the effects of hypoxia and reoxygenation in adult cardiomyocytes and the molecular mechanism involved in cardiomyocyte apoptosis. Morphologically, reoxygenation induced rounding up of the cells, appearance of membrane blebs that were filled with marginated mitochondria, and ultrastructural findings characteristic of apoptosis. Reoxygenation (18 hours of reoxygenation after 6 hours of hypoxia) and prolonged hypoxia (24 hours of hypoxia) resulted in a 59% and 51% decrease in cellular viability, respectively. During reoxygenation, cell death occurred predominantly via apoptosis associated with appearance of cytosolic cytochrome c and activation of caspase-3 and -9. However, nonapoptotic cell death predominated during prolonged hypoxia. Both caspase inhibition and Bcl-2 overexpression during reoxygenation significantly improved cellular viability through inhibition of apoptosis but had minimal effect on hypoxia-induced cell death. Bcl-2 overexpression blocked reoxygenation-induced cytochrome c release and activation of caspase -3 and -9, but caspase inhibition alone did not block cytochrome c release. These results suggest that apoptosis predominates in cardiomyocytes after reoxygenation through a mitochondrion-dependent apoptotic pathway, and Bcl-2 prevents reoxygenation-induced apoptosis by inhibiting cytochrome c release from the mitochondria and prevents activation of caspase-3 and -9. (Circ Res. 2000;87:118-125.)