© 2000 American Heart Association, Inc.
Cellular Biology |
Morphological and Molecular Characterization of Adult Cardiomyocyte Apoptosis During Hypoxia and Reoxygenation
From the Cardiovascular Division (P.M.K., A.H., H.A., S.I.) and Division of Cell Growth Regulation (A.U.), Beth Israel Deaconess Medical Center and Department of Medicine, Harvard Medical School, Boston, Mass.
Correspondence to Seigo Izumo, MD, Beth Israel Deaconess Medical Center, 330 Brookline Ave, SL-201, Boston, MA 02215. E-mail sizumo{at}caregroup.harvard.edu
Abstract—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.)
Key Words: caspases • Bcl-2 • cytochrome c • adenovirus • mitochondria
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|
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|
|
|
|
|
|
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|
|
|
|
|
|
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|
|
|
|
|
|
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|
|
|
|
|
|
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|
|
|
|
|
|
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|
|
|
|
|
|
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|
|
|
|
|
|
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|
|
|
|
|
|
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|
|
|
|
|
|
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|
|
|
|
|
|
L. Hauck, G. Hansmann, R. Dietz, and R. von Harsdorf Inhibition of Hypoxia-Induced Apoptosis by Modulation of Retinoblastoma Protein-Dependent Signaling in Cardiomyocytes Circ. Res., November 1, 2002; 91(9): 782 - 789. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 N. Bodyak, P. M. Kang, M. Hiromura, I. Sulijoadikusumo, N. Horikoshi, K. Khrapko, and A. Usheva Gene expression profiling of the aging mouse cardiac myocytes Nucleic Acids Res., September 1, 2002; 30(17): 3788 - 3794. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
W. Chao, Y. Shen, L. Li, and A. Rosenzweig Importance of FADD Signaling in Serum Deprivation- and Hypoxia-induced Cardiomyocyte Apoptosis J. Biol. Chem., August 23, 2002; 277(35): 31639 - 31645. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
Z.-Q. Zhao and J. Vinten-Johansen Myocardial apoptosis and ischemic preconditioning Cardiovasc Res, August 15, 2002; 55(3): 438 - 455. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. T. Crow Hypoxia, BNip3 Proteins, and the Mitochondrial Death Pathway in Cardiomyocytes Circ. Res., August 9, 2002; 91(3): 183 - 185. [Full Text] [PDF]
|
|
|
|
|
|
T. Date, A. J Belanger, S. Mochizuki, J. A Sullivan, L. X Liu, A. Scaria, S. H Cheng, R. J Gregory, and C. Jiang Adenovirus-mediated expression of p35 prevents hypoxia/reoxygenation injury by reducing reactive oxygen species and caspase activity Cardiovasc Res, August 1, 2002; 55(2): 309 - 319. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
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|
|
|
|
 |
|
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|
|
|
|
|
|
H. Aoki, P. M. Kang, J. Hampe, K. Yoshimura, T. Noma, M. Matsuzaki, and S. Izumo Direct Activation of Mitochondrial Apoptosis Machinery by c-Jun N-terminal Kinase in Adult Cardiac Myocytes J. Biol. Chem., March 15, 2002; 277(12): 10244 - 10250. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
C. Li and R. M. Jackson Reactive species mechanisms of cellular hypoxia-reoxygenation injury Am J Physiol Cell Physiol, February 1, 2002; 282(2): C227 - C241. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
D. S. McClintock, M. T. Santore, V. Y. Lee, J. Brunelle, G. R. S. Budinger, W.-X. Zong, C. B. Thompson, N. Hay, and N. S. Chandel Bcl-2 Family Members and Functional Electron Transport Chain Regulate Oxygen Deprivation-Induced Cell Death Mol. Cell. Biol., January 1, 2002; 22(1): 94 - 104. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
R. Malhotra, Z. Lin, C. Vincenz, and F. C. Brosius III Hypoxia induces apoptosis via two independent pathways in Jurkat cells: differential regulation by glucose Am J Physiol Cell Physiol, November 1, 2001; 281(5): C1596 - C1603. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
P. Narayan, R. M. Mentzer Jr., and R. D. Lasley Annexin V staining during reperfusion detects cardiomyocytes with unique properties Am J Physiol Heart Circ Physiol, November 1, 2001; 281(5): H1931 - H1937. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. N. Sharikabad, K. M. Ostbye, and O. Brors Increased [Mg2+]o reduces Ca2+ influx and disruption of mitochondrial membrane potential during reoxygenation Am J Physiol Heart Circ Physiol, November 1, 2001; 281(5): H2113 - H2123. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
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|
|
|
|
|
|
M. Xu, Y. Wang, A. Ayub, and M. Ashraf Mitochondrial KATP channel activation reduces anoxic injury by restoring mitochondrial membrane potential Am J Physiol Heart Circ Physiol, September 1, 2001; 281(3): H1295 - H1303. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 J. M. Pearl, D. P. Nelson, C. J. Wagner, J. P. Lombardi, and J. Y. Duffy Endothelin receptor blockade reduces ventricular dysfunction and injury after reoxygenation Ann. Thorac. Surg., August 1, 2001; 72(2): 565 - 570. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 R. Maruyama, G. Takemura, T. Aoyama, K. Hayakawa, M. Koda, Y. Kawase, X. Qiu, Y. Ohno, S. Minatoguchi, K. Miyata, et al. Dynamic Process of Apoptosis in Adult Rat Cardiomyocytes Analyzed Using 48-Hour Videomicroscopy and Electron Microscopy : Beating and Rate are Associated with the Apoptotic Process Am. J. Pathol., August 1, 2001; 159(2): 683 - 691. [Abstract] [Full Text]
|
|
|
|
|
|
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|
|
|
|
 |
|
 R. H. Chen, R. S. Farivar, and D. H. Adams Reply J. Thorac. Cardiovasc. Surg., February 1, 2001; 121(2): 401 - 401. [Full Text] [PDF]
|
|
|
|
|
|
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|
|