(Circulation. 2001;103:2617.)
© 2001 American Heart Association, Inc.
Basic Science Reports |
Mitochondria as Targets for Nitric Oxide–Induced Protection During Simulated Ischemia and Reoxygenation in Isolated Neonatal Cardiomyocytes
From the Departments of Cardiology, Kings College London, St Thomas’ Hospital (R.D.R., M.S.M.), and Physiology, University College London (M.H.M., M.R.D.), UK. The first 2 authors contributed equally to this work.
Correspondence to Michael R. Duchen, Department of Physiology, University College London, Gower St, London, WC1E 6BT, UK. E-mail m.duchen{at}ucl.ac.uk
Background—As shown previously, exposure to NO donors initiates protective mechanisms in cardiomyocytes that persist after removal of the donor, a form of pharmacological preconditioning. Because NO also affects mitochondrial respiration, we studied the effect of NO on mitochondrial Ca2+ uptake.
Methods and Results—Neonatal rat ventricular myocytes in primary culture were exposed to 1 hour of simulated ischemia and 1 hour of reoxygenation (sI/R). Pretreatment with the NO donor S-nitroso-N-acetyl-penicillamine (SNAP) (1 mmol/L for 90 minutes), followed by washing and incubation for 10 to 30 minutes, reduced sI/R-induced cell death to 25.4% compared with control (propidium iodide exclusion assay, P<0.001). Short (10-second) exposures to SNAP reversibly suppressed mitochondrial respiration without a detectable change in mitochondrial potential. In contrast, treatment with SNAP for 90 minutes caused a modest but sustained mitochondrial depolarization, as judged by JC-1 fluorescence. SNAP pretreatment limited cellular Ca2+ overload during ischemia (fura-2 ratio rose to 226±40% versus 516±170% of baseline, n=5, P<0.05) and prevented loss of cell membrane integrity during reoxygenation. SNAP pretreatment also significantly reduced the ability of mitochondria to accumulate Ca2+ in the face of a similar cytosolic Ca2+ load (peak rhod-2 fluorescence 133±4% versus 166±7% of baseline at similar fluo-3 levels, P=0.0004, n=52 and 25, respectively).
Conclusions—Pretreatment with an NO donor induces a modest, sustained mitochondrial depolarization and protects cardiomyocytes from sI/R injury. The demonstrated reduction in mitochondrial Ca2+ uptake possibly reduces cytosolic Ca2+ overload, providing a likely mechanism for NO-induced protection.
Key Words: nitric oxide • cells • calcium • ischemia • reperfusion
This article has been cited by other articles:
 |
|
 |
|
  M.-G. Ryou, J. Sun, K. N. Oguayo, E. B. Manukhina, H. F. Downey, and R. T. Mallet Hypoxic Conditioning Suppresses Nitric Oxide Production upon Myocardial Reperfusion Experimental Biology and Medicine, June 1, 2008; 233(6): 766 - 774. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 S. Shiva, M. N. Sack, J. J. Greer, M. Duranski, L. A. Ringwood, L. Burwell, X. Wang, P. H. MacArthur, A. Shoja, N. Raghavachari, et al. Nitrite augments tolerance to ischemia/reperfusion injury via the modulation of mitochondrial electron transfer J. Exp. Med., September 3, 2007; 204(9): 2089 - 2102. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 A. Caplanusi, A. J. Fuller, R. A. Gonzalez-Villalobos, T. G. Hammond, and L. G. Navar Metabolic inhibition-induced transient Ca2+ increase depends on mitochondria in a human proximal renal cell line Am J Physiol Renal Physiol, August 1, 2007; 293(2): F533 - F540. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 H. Gao, L. Chen, and H.-T. Yang Activation of {alpha}1B-adrenoceptors alleviates ischemia/reperfusion injury by limitation of mitochondrial Ca2+ overload in cardiomyocytes Cardiovasc Res, August 1, 2007; 75(3): 584 - 595. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
C. Dezfulian, N. Raat, S. Shiva, and M. T. Gladwin Role of the anion nitrite in ischemia-reperfusion cytoprotection and therapeutics Cardiovasc Res, July 15, 2007; 75(2): 327 - 338. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
Y. Jang, H. Wang, J. Xi, R. A. Mueller, E. A. Norfleet, and Z. Xu NO mobilizes intracellular Zn2+ via cGMP/PKG signaling pathway and prevents mitochondrial oxidant damage in cardiomyocytes Cardiovasc Res, July 15, 2007; 75(2): 426 - 433. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
C. Martin, R. Schulz, H. Post, K. Boengler, M. Kelm, P. Kleinbongard, P. Gres, A. Skyschally, I. Konietzka, and G. Heusch Microdialysis-based analysis of interstitial NO in situ: NO synthase-independent NO formation during myocardial ischemia Cardiovasc Res, April 1, 2007; 74(1): 46 - 55. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. N. Sack Mitochondrial depolarization and the role of uncoupling proteins in ischemia tolerance Cardiovasc Res, November 1, 2006; 72(2): 210 - 219. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 M. N. Sack Exploring Mitochondria in the Intact Ischemic Heart: Advancing Technologies to Image Intracellular Function Circulation, October 3, 2006; 114(14): 1452 - 1454. [Full Text] [PDF]
|
|
|
|
|
|
S. M. Davidson and M. R. Duchen Effects of NO on mitochondrial function in cardiomyocytes: Pathophysiological relevance Cardiovasc Res, July 1, 2006; 71(1): 10 - 21. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
N. Smart, M. H. Mojet, D. S. Latchman, M. S. Marber, M. R. Duchen, and R. J. Heads IL-6 induces PI 3-kinase and nitric oxide-dependent protection and preserves mitochondrial function in cardiomyocytes Cardiovasc Res, January 1, 2006; 69(1): 164 - 177. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 S. M. Davidson and D. M. Yellon The role of nitric oxide in mitochondria. Focus on "Modulation of mitochondrial Ca2+ by nitric oxide in cultured bovine vascular endothelial cells" Am J Physiol Cell Physiol, October 1, 2005; 289(4): C775 - C777. [Full Text] [PDF]
|
|
|
|
|
|
E. N. Dedkova and L. A. Blatter Modulation of mitochondrial Ca2+ by nitric oxide in cultured bovine vascular endothelial cells Am J Physiol Cell Physiol, October 1, 2005; 289(4): C836 - C845. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 J. Chen, J. X. Zhu, I. Wilson, and J. S. Cameron Cardioprotective effects of KATP channel activation during hypoxia in goldfish Carassius auratus J. Exp. Biol., July 15, 2005; 208(14): 2765 - 2772. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 J. D. Marks, C. Boriboun, and J. Wang Mitochondrial Nitric Oxide Mediates Decreased Vulnerability of Hippocampal Neurons from Immature Animals to NMDA J. Neurosci., July 13, 2005; 25(28): 6561 - 6575. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 H.-Y. Sun, N.-P. Wang, F. Kerendi, M. Halkos, H. Kin, R. A. Guyton, J. Vinten-Johansen, and Z.-Q. Zhao Hypoxic postconditioning reduces cardiomyocyte loss by inhibiting ROS generation and intracellular Ca2+ overload Am J Physiol Heart Circ Physiol, April 1, 2005; 288(4): H1900 - H1908. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
Z. Xu, S.-S. Park, R. A. Mueller, R. C. Bagnell, C. Patterson, and P. G. Boysen Adenosine produces nitric oxide and prevents mitochondrial oxidant damage in rat cardiomyocytes Cardiovasc Res, March 1, 2005; 65(4): 803 - 812. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
G. Wang, D. A. Liem, T. M. Vondriska, H. M. Honda, P. Korge, D. M. Pantaleon, X. Qiao, Y. Wang, J. N. Weiss, and P. Ping Nitric oxide donors protect murine myocardium against infarction via modulation of mitochondrial permeability transition Am J Physiol Heart Circ Physiol, March 1, 2005; 288(3): H1290 - H1295. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
I. Smets, A. Caplanusi, S. Despa, Z. Molnar, M. Radu, M. vandeVen, M. Ameloot, and P. Steels Ca2+ uptake in mitochondria occurs via the reverse action of the Na+/Ca2+ exchanger in metabolically inhibited MDCK cells Am J Physiol Renal Physiol, April 1, 2004; 286(4): F784 - F794. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
Z. Xu, X. Ji, and P. G. Boysen Exogenous nitric oxide generates ROS and induces cardioprotection: involvement of PKG, mitochondrial KATP channels, and ERK Am J Physiol Heart Circ Physiol, April 1, 2004; 286(4): H1433 - H1440. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
R. Schulz, M. Kelm, and G. Heusch Nitric oxide in myocardial ischemia/reperfusion injury Cardiovasc Res, February 15, 2004; 61(3): 402 - 413. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
F. Eefting, B. Rensing, J. Wigman, W. J. Pannekoek, W. M. Liu, M. J. Cramer, D. J Lips, and P. A Doevendans Role of apoptosis in reperfusion injury Cardiovasc Res, February 15, 2004; 61(3): 414 - 426. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
G. C Rodrigo, N. W Davies, and N. B Standen Diazoxide causes early activation of cardiac sarcolemmal KATP channels during metabolic inhibition by an indirect mechanism Cardiovasc Res, February 15, 2004; 61(3): 570 - 579. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
E. N. Dedkova, X. Ji, S. L. Lipsius, and L. A. Blatter Mitochondrial calcium uptake stimulates nitric oxide production in mitochondria of bovine vascular endothelial cells Am J Physiol Cell Physiol, February 1, 2004; 286(2): C406 - C415. [Abstract] [Full Text]
|
|
|
|
 |
|
 L. SCHILD, T. REINHECKEL, M. REISER, T. F. W. HORN, G. WOLF, and W. AUGUSTIN Nitric oxide produced in rat liver mitochondria causes oxidative stress and impairment of respiration after transient hypoxia FASEB J, December 1, 2003; 17(15): 2194 - 2201. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. S. Cameron, K. E. Hoffmann, C. Zia, H. M. Hemmett, A. Kronsteiner, and C. M. Lee A role for nitric oxide in hypoxia-induced activation of cardiac KATP channels in goldfish (Carassius auratus) J. Exp. Biol., November 15, 2003; 206(22): 4057 - 4065. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
Y. Maejima, S. Adachi, H. Ito, K. Nobori, M. Tamamori-Adachi, and M. Isobe Nitric oxide inhibits ischemia/reperfusion-induced myocardial apoptosis by modulating cyclin A-associated kinase activity Cardiovasc Res, August 1, 2003; 59(2): 308 - 320. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
P. P. Dzeja, P. Bast, C. Ozcan, A. Valverde, E. L. Holmuhamedov, D. G. L. Van Wylen, and A. Terzic Targeting nucleotide-requiring enzymes: implications for diazoxide-induced cardioprotection Am J Physiol Heart Circ Physiol, April 1, 2003; 284(4): H1048 - H1056. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 R. J. Hassink, A. Brutel de la Riviere, C. L. Mummery, and P. A. Doevendans Transplantation of cells for cardiac repair J. Am. Coll. Cardiol., March 5, 2003; 41(5): 711 - 717. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
R. M Bell, H. L Maddock, and D. M Yellon The cardioprotective and mitochondrial depolarising properties of exogenous nitric oxide in mouse heart Cardiovasc Res, February 1, 2003; 57(2): 405 - 415. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
B. Billups and I. D. Forsythe Presynaptic Mitochondrial Calcium Sequestration Influences Transmission at Mammalian Central Synapses J. Neurosci., July 15, 2002; 22(14): 5840 - 5847. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 P. P. Dzeja, E. L. Holmuhamedov, C. Ozcan, D. Pucar, A. Jahangir, and A. Terzic Mitochondria: Gateway for Cytoprotection Circ. Res., October 26, 2001; 89(9): 744 - 746. [Full Text] [PDF]
|
|
|
|
 |
|
 R D Rakhit and M S Marber Nitric oxide: an emerging role in cardioprotection? Heart, October 1, 2001; 86(4): 368 - 372. [Full Text] [PDF]
|
|
|
|
|
|
J. Minners, L. Lacerda, J. McCarthy, J. J. Meiring, D. M. Yellon, and M. N. Sack Ischemic and Pharmacological Preconditioning in Girardi Cells and C2C12 Myotubes Induce Mitochondrial Uncoupling Circ. Res., October 26, 2001; 89(9): 787 - 792. [Abstract] [Full Text] [PDF]
|
|