This Article Full Text Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Request Permissions Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Wu, M.-L. Articles by Lee, Y.-T. Search for Related Content PubMed PubMed Citation Articles by Wu, M.-L. Articles by Lee, Y.-T. Pubmed/NCBI databases Compound via MeSH Substance via MeSH Hazardous Substances DB CALCIUM COMPOUNDS CALCIUM, ELEMENTAL HYDROGEN PEROXIDE

(Circulation Research. 1996;78:564-572.)
© 1996 American Heart Association, Inc.

Articles

Mechanism of Hydrogen Peroxide and Hydroxyl Free Radical–Induced Intracellular Acidification in Cultured Rat Cardiac Myoblasts

Mei-Lin Wu, Ke-Li Tsai, Seu-Mei Wang, Jiahn-Chun Wu, Bor-Sen Wang, Yuan-Teh Lee

From the Departments of Physiology (M.-L.W., K.-L.T.) and Anatomy (S.-M.W., J.-C.W.), the Institute of Toxicology (B.-S.W.), and the Department of Internal Medicine (Y.-T.L.), Center for Cardiovascular Research, College of Medicine, National Taiwan University, Taipei, ROC.

Correspondence to Dr Yuan-Teh Lee, Department of Internal Medicine, Medical College, National Taiwan University Hospital, 7, Chung-Shan South Rd, Taipei, Taiwan, ROC.

Abstract After a transient ischemic attack of the cardiac vascular system, reactive oxygen-derived free radicals, including the superoxide (O₂^-) and hydroxyl (OH) radicals can be easily produced during reperfusion. These free radicals have been suggested to be responsible for reperfusion-induced cardiac stunning and reperfusion-induced arrhythmia. Hydrogen peroxide (H₂O₂) is often used as an experimental source of oxygen-derived free radicals. Using freshly dissociated single rat cardiac myocytes and the rat cardiac myoblast cell line, H9c2, we have shown, for the first time, that an intriguing pH_i acidification (0.24 pH unit) is induced by the addition of 100 µmol/L H₂O₂ and that this dose is without effect on the intracellular free Ca²⁺ levels or viability of the cells. Using H9c2 as a model cardiac cell, we have shown that it is the intracellular production of OH, and not O₂^- or H₂O₂, that results in this acidification. We have excluded any involvement of (1) the three known cardiac pH_i regulators (the Na⁺-H⁺ exchanger, the Cl^--HCO₃ exchanger, and the Na⁺-HCO₃ cotransporter), (2) a rise in intracellular Ca²⁺ levels, and (3) inhibition of oxidative phosphorylation. However, we have found that H₂O₂-induced acidosis is due to inhibition of the glycolytic pathway, with hydrolysis of intracellular ATP and the resultant intracellular acidification. In cardiac muscle and in skinned cardiac muscle fiber, it has been shown that a small intracellular acidification may severely inhibit contractility. Therefore, the sustained pH_i decrease caused by hydroxyl radicals may contribute, in some part, to the well-documented impairment of cardiac mechanical function (ie, reperfusion cardiac stunning) seen during reperfusion ischemia.

Key Words: cardiac myocytes • hydroxyl free radical • reperfusion cardiac stunning • intracellular acidosis • rat H9c2 cardiac cell line

This article has been cited by other articles:

				D. A. Kubli, M. N. Quinsay, C. Huang, Y. Lee, and A. B. Gustafsson Bnip3 functions as a mitochondrial sensor of oxidative stress during myocardial ischemia and reperfusion Am J Physiol Heart Circ Physiol, November 1, 2008; 295(5): H2025 - H2031. [Abstract] [Full Text] [PDF]

				Y. Miyamoto, T. Saiwaki, J. Yamashita, Y. Yasuda, I. Kotera, S. Shibata, M. Shigeta, Y. Hiraoka, T. Haraguchi, and Y. Yoneda Cellular stresses induce the nuclear accumulation of importin {alpha} and cause a conventional nuclear import block J. Cell Biol., June 7, 2004; 165(5): 617 - 623. [Abstract] [Full Text] [PDF]

				D. K. Mulkey, R. A. Henderson III, N. A. Ritucci, R. W. Putnam, and J. B. Dean Oxidative stress decreases pHi and Na+/H+ exchange and increases excitability of solitary complex neurons from rat brain slices Am J Physiol Cell Physiol, April 1, 2004; 286(4): C940 - C951. [Abstract] [Full Text] [PDF]

				D. K. Mulkey, R. A. Henderson III, R. W. Putnam, and J. B. Dean Hyperbaric oxygen and chemical oxidants stimulate CO2/H+-sensitive neurons in rat brain stem slices J Appl Physiol, September 1, 2003; 95(3): 910 - 921. [Abstract] [Full Text] [PDF]

				E. B. Affar, R. G. Shah, A.-K. Dallaire, V. Castonguay, and G. M. Shah Role of poly(ADP-ribose) polymerase in rapid intracellular acidification induced by alkylating DNA damage PNAS, December 21, 2001; (2001) 12460399. [Abstract] [Full Text] [PDF]

				T. R. Krieger and R. Loch-Caruso Antioxidants Prevent {{gamma}}-Hexachlorocyclohexane-Induced Inhibition of Rat Myometrial Gap Junctions and Contractions Biol Reprod, February 1, 2001; 64(2): 537 - 547. [Abstract] [Full Text]

				M.-L. Wu, C.-C. Chan, and M.-J. Su Possible Mechanism(s) of Arachidonic Acid-Induced Intracellular Acidosis in Rat Cardiac Myocytes Circ. Res., February 18, 2000; 86 (3): e55 - e62. [Abstract] [Full Text] [PDF]

				P. Lipton Ischemic Cell Death in Brain Neurons Physiol Rev, October 1, 1999; 79(4): 1431 - 1568. [Abstract] [Full Text] [PDF]

				D. A. Siwik, J. D. Tzortzis, D. R. Pimental, D. L.-F. Chang, P. J. Pagano, K. Singh, D. B. Sawyer, and W. S. Colucci Inhibition of Copper-Zinc Superoxide Dismutase Induces Cell Growth, Hypertrophic Phenotype, and Apoptosis in Neonatal Rat Cardiac Myocytes In Vitro Circ. Res., July 23, 1999; 85(2): 147 - 153. [Abstract] [Full Text] [PDF]

				W. Wang, M. Watanabe, T. Nakamura, Y. Kudo, and R. Ochi Properties and expression of Ca2+-activated K+ channels in H9c2 cells derived from rat ventricle Am J Physiol Heart Circ Physiol, May 1, 1999; 276(5): H1559 - H1566. [Abstract] [Full Text] [PDF]

				Q. Hu, Y. Xia, S. Corda, J. L. Zweier, and R. C. Ziegelstein Hydrogen Peroxide Decreases pHi in Human Aortic Endothelial Cells by Inhibiting Na+/H+ Exchange Circ. Res., September 21, 1998; 83(6): 644 - 651. [Abstract] [Full Text] [PDF]

				J. I. Kourie Interaction of reactive oxygen species with ion transport mechanisms Am J Physiol Cell Physiol, July 1, 1998; 275(1): C1 - C24. [Abstract] [Full Text] [PDF]

				A. Sabri, K. L. Byron, A. M. Samarel, J. Bell, and P. A. Lucchesi Hydrogen Peroxide Activates Mitogen-Activated Protein Kinases and Na+-H+ Exchange in Neonatal Rat Cardiac Myocytes Circ. Res., June 1, 1998; 82(10): 1053 - 1062. [Abstract] [Full Text] [PDF]

				Y.-S. Liaw, P.-C. Yang, C.-J. Yu, S.-H. Kuo, K.-T. Luh, Y.-J. Lin, and M.-L. Wu PKC activation is required by EGF-stimulated Na+-H+ exchanger in human pleural mesothelial cells Am J Physiol Lung Cell Mol Physiol, May 1, 1998; 274(5): L665 - L672. [Abstract] [Full Text] [PDF]

				J. L. Hirpara, M.-V. Clement, and S. Pervaiz Intracellular Acidification Triggered by Mitochondrial-derived Hydrogen Peroxide Is an Effector Mechanism for Drug-induced Apoptosis in Tumor Cells J. Biol. Chem., January 5, 2001; 276(1): 514 - 521. [Abstract] [Full Text] [PDF]

				E. B. Affar, R. G. Shah, A.-K. Dallaire, V. Castonguay, and G. M. Shah Role of poly(ADP-ribose) polymerase in rapid intracellular acidification induced by alkylating DNA damage PNAS, January 8, 2002; 99(1): 245 - 250. [Abstract] [Full Text] [PDF]

				M. Cougnon, S. Benammou, F. Brouillard, P. Hulin, and G. Planelles Effect of reactive oxygen species on NH4+ permeation in Xenopus laevis oocytes Am J Physiol Cell Physiol, June 1, 2002; 282(6): C1445 - C1453. [Abstract] [Full Text] [PDF]