TPEN, a Specific Zn2+ Chelator, Inhibits Sodium Dithionite and Glucose Deprivation (SDGD)-Induced Neuronal Death by Modulating Apoptosis, Glutamate Signaling, and Voltage-Gated K+ and Na+ Channels
Hypoxia–ischemia-induced neuronal death is an important pathophysiological process that accompanies ischemic stroke and represents a major challenge in preventing ischemic stroke. To elucidate factors related to and a potential preventative mechanism of hypoxia–ischemia-induced neuronal death, primary neurons were exposed to sodium dithionite and glucose deprivation (SDGD) to mimic hypoxic–ischemic conditions. The effects of N,N,N′,N′-tetrakis (2-pyridylmethyl) ethylenediamine (TPEN), a specific Zn2+-chelating agent, on SDGD-induced neuronal death, glutamate signaling (including the free glutamate concentration and expression of α-amino-3-hydroxy-5-methyl-4-isoxazolepropionate (AMPA) receptor (GluR2) and N-methyl-d-aspartate (NMDA) receptor subunits (NR2B), and voltage-dependent K+ and Na+ channel currents were also investigated. Our results demonstrated that TPEN significantly suppressed increases in cell death, apoptosis, neuronal glutamate release into the culture medium, NR2B protein expression, and I K as well as decreased GluR2 protein expression and Na+ channel activity in primary cultured neurons exposed to SDGD. These results suggest that TPEN could inhibit SDGD-induced neuronal death by modulating apoptosis, glutamate signaling (via ligand-gated channels such as AMPA and NMDA receptors), and voltage-gated K+ and Na+ channels in neurons. Hence, Zn2+ chelation might be a promising approach for counteracting the neuronal loss caused by transient global ischemia. Moreover, TPEN could represent a potential cell-targeted therapy.
KeywordsSodium dithionite Glucose deprivation TPEN Neuronal death Glutamate signal path Voltage-gated channel
This work was supported by the National Natural Science Foundation of China (No. 31272317), the Natural Science Foundation of Tianjin City (15JCYBJC24500), and the 111 Project (B08011).
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Conflicts of interest
There are no conflicts of interest to declare.
- Frederickson CJ, Giblin LJ, Krezel A, McAdoo DJ, Muelle RN, Zeng Y, Balaji RV, Masalha R, Thompson RB, Fierke CA (2006) Concentrations of extracellular free zinc (pZn)(e) in the central nervous system during simple anesthetization, ischemia and reperfusion. Exp Neurol 198(2):285–293CrossRefPubMedGoogle Scholar
- Gill R, Alanine A, Bourson A, Buttelmann B, Fischer G, Heitz MP, Kew JNC, Levet-Trafit B, Lorez HP, Malherbe P (2002) Pharmacological characterization of Ro 63-1908 (1-2-(4-hydroxy-phenoxy) -ethyl-4-(4-methyl-benzyl)-piperidin-4-ol), a novel subtype-selective N-methyl-d-aspartate antagonist. J Pharmacol Exp Ther 302(3):940–948CrossRefPubMedGoogle Scholar
- Heimlich G, Bortner CD, Cidlowski JA (2004) Cell volume and signaling. In: Lauf PK, Adragna NC (eds) Apoptosis and cell volume regulation: the importance of ions and ion channels. Springer, New York, pp 189–203Google Scholar
- Hetz C, Vitte PA, Bombrun A, Rostovtseva TK, Montessuit S, Hiver A, Schwarz MK, Church DJ, Korsmeyer SJ, Martinou JC (2005) Bax channel inhibitors prevent mitochondrion-mediated apoptosis and protect neurons in a model of global brain ischemia. J Biol Chem 280(52):42960–42970CrossRefPubMedGoogle Scholar
- Paul D, Saias L, Pedinotti JC, Chabert M, Magnifico S, Pallandre A, De Lambert B, Houdayer C, Brugg B, Peyrin JM, Viovy JL (2011) A “dry and wet hybrid” lithography technique for multilevel replication templates: applications to microfluidic neuron culture and two-phase global mixing. Biomicrofluidics 5:24102CrossRefPubMedGoogle Scholar
- Wang WM, Liu Z, Liu AJ, Wang YX, Wang HG, An D, Heng B, Xie LH, Duan JL, Liu YQ (2015) The zinc ion chelating agent TPEN attenuates neuronal death/apoptosis caused by hypoxia/ischemia via mediating the pathophysiological cascade including excitotoxicity, oxidative stress, and inflammation. CNS Neurosci Ther 21(9):708–717CrossRefPubMedGoogle Scholar