Abstract
Oxidative stress and nitrosative stress are both suggested to be involved in cardiac ischemia-reperfusion (I/R) injury. Using time-lapse confocal microscopy of cardiomyocytes and high-affinity O2−• and Zn2+ probes, this study is the first to show that I/R, reactive oxygen species (ROS), and reactive nitrogen species (RNS) all cause a marked increase in the [O2−•]i, resulting in cytosolic and mitochondrial Zn2+ release. Exposure to a cell-penetrating, high-affinity Zn2+i chelator, TPEN, largely abolished the Zn2+i release and markedly protected myocytes from I/R-, ROS-, RNS-, or Zn2+/K+ (Zn2+i supplementation)-induced myocyte apoptosis for at least 24 h after TPEN removal. Flavonoids and U0126 (a MEK1/2 inhibitor) largely inhibited the myocyte apoptosis and the TPEN-sensitive I/R- or Zn2+i supplement-induced persistent extracellular signal-regulated kinase 1 and 2 (ERK1/2) phosphorylation, dephosphorylation of p-Ser9 on glycogen synthase kinase 3β (GSK-3β), and the translocation into and accumulation of p-Tyr216 GSK-3β and p53 in, the nucleus. Silencing of GSK-3β or p53 expression was cardioprotective, indicating that activation of the ERK–GSK-3β–p53 signaling pathway is involved in Zn2+-sensitive myocyte death. Moreover, the ERK-dependent Noxa–myeloid cell leukemia-1 (Mcl-1) pathway is also involved, as silencing of Noxa expression was cardioprotective and U0126 abolished both the increase in Noxa expression and in Mcl-1 degradation. Thus, acute upstream Zn2+i chelation at the start of reperfusion and the use of natural products, that is, flavonoids, may be beneficial in the treatment of cardiac I/R injury.
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Abbreviations
- I/R injury:
-
ischemia/reperfusion injury
- ROS:
-
reactive oxygen species
- RNS:
-
reactive nitrogen species
- Zn2+i:
-
intracellular free Zn2+ ions
- TPEN, N,N,N′,N′:
-
tetrakis (2-pyridylmethyl) ethylenediaminepentaethylene
- NAC:
-
N-acetylcysteine
- DTT:
-
dithiothreitol
- MT:
-
metallothionein
- mPTP:
-
mitochondrial permeability transition pore
- PFT-α:
-
pifithrin-α
- MEK1/2:
-
kinase mitogen protein kinase kinase 1 and 2
- ERK1/2:
-
extracellular signal-regulated kinase 1 and 2
- GSK-3β:
-
glycogen synthase kinase-3β
- Mcl-1:
-
myeloid cell leukemia-1
- LOX:
-
lipoxygenase
- NFκB:
-
nuclear factor κB
- HIF1α:
-
hypoxia-inducible factor-1
- PKC:
-
protein kinase C
References
Maret W, Vallee BL . Thiolate ligands in metallothionein confor redox activity on Zn2+ chelators. Proc Natl Acad Sci 1998; 95: 3478–3482.
Bozym RA, Thompson RB, Stoddard AK, Fierke CA . Measuring picomolar intracellular exchangeable zinc in PC-12 cells using a ratiometric fluorescence biosensor. ACS Chem Biol 2006; 1: 103–111.
Cheung P-Y, Wang W, Schulz R . Glutathione protects against myocardial ischemia-reperfusion injury by detoxifying peroxynitrite. J Mol Cell Cardiol 2000; 32: 1669–1678.
Juhaszova M, Zorov DB, Kim S-H, Pepe S, Fu Q, Fishbein KW et al. Glycogen synthase kinase-3β mediates convergence of protection signaling to inhibit the mitochondrial permeability transition pore. J Clin Invest 2004; 113: 1535–1549.
Yellon DM, Hausenloy DJ . Myocardial Reperfusion Injury. N Engl J Med 2007; 357: 1121–1135.
Kang YJ, Li G, Saari JT . Metallothionein inhibits ischemia-reperfusion injury in mouse heart. Am J Physiol 1999; 26: H993–H997.
Bossy-Wetzel E, Talantova MV, Lee WD, Schölzke MN, Harrop A, Mathews E et al. Crosstalk between nitric oxide and zinc pathways to neuronal cell death involving mitochondrial dysfunction and p38-activated K+ channels. Neuron 2004; 41: 351–365.
Zhang Y, Wang H, Li J, Dong L, Xu P, Chen W et al. Intracellular zinc release and ERK phosphorylation are required upstream of 12-lipoxygenease activation in peroxynitrite toxicity to mature rat oligodendrocytes. J Biol Chem 2006; 281: 9460–9470.
Mclntosh R, Lee SY, Ghio AJ, Xi J, Zhu M, Shen X et al. The critical role of intracellular zinc in adenosine A2 receptor activation induced cardioprotection against reperfusion injury. J Mol Cell Cardiol 2010; 49: 41–47.
Karagulova G, Yue Y, Moreyra A, Boutjdir M, Korichneva I . Protective role of intracellular zinc in myocardial ischemia/reperfusion is associated with preservation of protein kinase C isoforms. JPET 2007; 321: 517–525.
Sensi SL, Ton-That D, Sullivan PG, Jonas EA, Gee KR, Kaczmarek LK et al. Modulation of mitochondrial function by endogenous Zn2+ pools. Proc Natl Acad Sci 2003; 100: 6157–6162.
Cornell NW, Crivaro KE . Stability constant for the zinc-dithiothreitol complex. Analy Biochem 1972; 47: 203–208.
Das S, Wong R, Rajapakse N, Murphy E, Steenbergen C . Glycogen synthase kinase 3 inhibition slows mitochondrial adenine nucleotide transport and regulates voltage-dependent anion channel phosphorylation. Circ Res 2008; 103: 983–991.
Gross ER, Hsu AK, Gross GJ . Opioid-Induced cardioprotection occurs via glycogen synthase Kinase β inhibition during reperfusion in intact rat hearts. Circ Res 2004; 94: 960–966.
Gomez L, Paillard M, Thibault H, Derumeaux G, Ovize M . Inhibition of GSK-3β by postconditioning is required to prevent opening of the mitochondrial permeability transition pore during reperfusion. Circulation 2008; 117: 2761–2768.
Miura T, Miki T . GSK-3β, a therapeutic target for cardiomyocyte protection. Circ J 2009; 73: 1184–1192.
Yook JI, Li X-Y, Ota I, Hu C, Kim HS, Kim NH et al. A Wnt–Axin2–GSK3β cascade regulates Snail1 activity in breast cancer cells. Nat Cell Biol 2006; 8: 1398–1406.
Beurel E, Jope BS . The paradoxical pro-and anti-apoptotic actions of GSK3 in the intrinsic and extrinsic apoptosis signaling pathways. Prog in Neurobiol 2006; 79: 173–189.
Lin T, Mak NK, Yang MS . MAPK regulate p53-dependent cell death induced by benzo[a]pyrene: involvement of p53 phosphorylation and acetylation. Toxicology 2008; 247: 145–153.
Taylor RC, Cullen SP, Martin SJ . Apoptosis: controlled demolition at the cellular level. Nat Rev Mol Cell Biol 2008; 9: 231–241.
Sheridan C, Brumatti G, Elgendy M, Brunet M, Martin SJ . An ERK-dependent pathway to Noxa expression regulates apoptosis by platinum-based chemotherapeutic drugs. Oncogene 2010; 29: 6428–6441.
Aikawa T, Shinzawa K, Tanaka N, Tsujimoto Y . Noxa is necessary for hydrogen peroxide-induced caspase-dependent cell death. FEBS Letters 2010; 584: 681–688.
Chanoit G, Lee SR, Xi J, Zhu M, McIntosh RA, Mueller RA et al. Exogenous zinc protects cardiac cells from reperfusion injury by targeting mitochondrial permeability transition pore through inactivation of glycogen synthase kinase-3β. Am J Physiol 2008; 295: H1227–H1233.
Lee SY, Chanoit G, McIntosh R, Zvara DA, Xu Z . Molecular mechanism underlying Akt activation in zinc-induced cardioprotection. Am J Physiol 2009; 297: H569–H575.
Eom TY, Roth KA, Jope RS . Neural precursor cells are protected from apoptosis induced by trophic factor withdrawal or genotoxic stress by inhibitors of glycogen synthase kinase 3. J Biol Chem 2007; 282: 22856–22864.
Cagnol S, Chambard J-C . ERK and cell death: mechanisms of ERK-induced cell death–apoptosis, autophagy and senescence. FEBS J 2010; 277: 2–21.
Kohda Y, Matsunaga Y, Shiota R, Satoh T, Kishi Y, Kawai Y et al. Raf-1/Mek/ERK1/2 signaling pathway in zinc-induced injury in rat renal cortical slices. J Toxicol Sci 2006; 31: 207–217.
Maurer U, Charvet C, Wagman AS, Dejardin E, Green DR . Glycogen synthase kinase-3 regulates mitochondrial outer membrane permeabilization and apoptosis by destabilization of Mcl-1. Mol Cell 2006; 21: 749–760.
Ding Q, He X, Hsu J-M, Xia W, Chen C-T, Li L-Y et al. Degradation of Mcl-1 by β-TrCP mediates glycogen synthase kinase 3-induced tumor suppression and chemosensitization. Mol Cell Biol 2007; 27: 4006–4017.
Frederickson CJ, Burdette SC, Frederickson CJ, Sensi SL, Weiss JH, Yin HZ et al. Method for identifying neuronal cells suffering zinc toxicity by use of a novel fluorescent sensor. J Neurosci Methods 2004; 139: 79–89.
Acknowledgements
We gratefully thank the National Science Council of Taiwan for financial support (grants: NSC-98-2320-B-002-042-MY3 and NSC 99-2314-B-166-001-MY3). We also gratefully acknowledge the Second Core Laboratory of Department of Medical Research, National Taiwan University Hospital for expert technical assistance.
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Lin, CL., Tseng, HC., Chen, WP. et al. Intracellular zinc release-activated ERK-dependent GSK-3β–p53 and Noxa–Mcl-1 signaling are both involved in cardiac ischemic-reperfusion injury. Cell Death Differ 18, 1651–1663 (2011). https://doi.org/10.1038/cdd.2011.80
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DOI: https://doi.org/10.1038/cdd.2011.80
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