Department of Anesthesiology and Critical Care Medicine (C.L., C.I.S., H.B., M.H.) and Department of General Neurosurgery, Cellular Neurophysiology (T.G.), University Medical Center Freiburg, Freiburg, Germany; and Department of Anatomy and Cell Biology (S.Z., B.H.) and Pharmaceutical Bioinformatics, Institute of Pharmaceutical Sciences (A.E.), Albert-Ludwigs-University of Freiburg, Freiburg, Germany.
J Pharmacol Exp Ther. 2013 Dec;347(3):781-93. doi: 10.1124/jpet.113.205989. Epub 2013 Sep 18.
Oxygen deprivation during ischemic or hemorrhagic stroke results in ATP depletion, loss of ion homeostasis, membrane depolarization, and excitotoxicity. Pharmacologic restoration of cellular energy supply may offer a promising concept to reduce hypoxic cell injury. In this study, we investigated whether carbimazole, a thionamide used to treat hyperthyroidism, reduces neuronal cell damage in oxygen-deprived human SK-N-SH cells or primary cortical neurons. Our results revealed that carbimazole induces an inhibitory phosphorylation of eukaryotic elongation factor 2 (eEF2) that was associated with a marked inhibition of global protein synthesis. Translational inhibition resulted in significant bioenergetic savings, preserving intracellular ATP content in oxygen-deprived neuronal cells and diminishing hypoxic cellular damage. Phosphorylation of eEF2 was mediated by AMP-activated protein kinase and eEF2 kinase. Carbimazole also induced a moderate calcium influx and a transient cAMP increase. To test whether translational inhibition generally diminishes hypoxic cell damage when ATP availability is limiting, the translational repressors cycloheximide and anisomycin were used. Cycloheximide and anisomycin also preserved ATP content in hypoxic SK-N-SH cells and significantly reduced hypoxic neuronal cell damage. Taken together, these data support a causal relation between the pharmacologic inhibition of global protein synthesis and efficient protection of neurons from ischemic damage by preservation of high-energy metabolites in oxygen-deprived cells. Furthermore, our results indicate that carbimazole or other translational inhibitors may be interesting candidates for the development of new organ-protective compounds. Their chemical structure may be used for computer-assisted drug design or screening of compounds to find new agents with the potential to diminish neuronal damage under ATP-limited conditions.
缺血性或出血性中风期间的缺氧会导致 ATP 耗竭、离子动态平衡丧失、膜去极化和兴奋性毒性。细胞能量供应的药理学恢复可能提供一个有前途的概念,以减少缺氧细胞损伤。在这项研究中,我们研究了甲状腺功能亢进的治疗药物甲巯咪唑是否可以减少缺氧的人 SK-N-SH 细胞或原代皮质神经元中的神经细胞损伤。我们的结果表明,甲巯咪唑诱导真核延伸因子 2(eEF2)的抑制性磷酸化,这与全局蛋白质合成的显著抑制有关。翻译抑制导致显著的能量节约,在缺氧神经元细胞中保留细胞内 ATP 含量,并减少缺氧细胞损伤。eEF2 的磷酸化由 AMP 激活的蛋白激酶和 eEF2 激酶介导。甲巯咪唑还诱导适度的钙内流和短暂的 cAMP 增加。为了测试当 ATP 可用性有限时翻译抑制是否普遍减少缺氧细胞损伤,使用了翻译抑制剂环己酰亚胺和放线菌酮。环己酰亚胺和放线菌酮也可在缺氧 SK-N-SH 细胞中保留 ATP 含量,并显著减少缺氧神经元细胞损伤。总之,这些数据支持在缺氧细胞中通过保存高能代谢物来抑制全局蛋白质合成和有效保护神经元免受缺血性损伤之间存在因果关系。此外,我们的结果表明,甲巯咪唑或其他翻译抑制剂可能是开发新的器官保护化合物的有趣候选物。它们的化学结构可用于计算机辅助药物设计或筛选化合物,以寻找具有在 ATP 有限条件下减少神经元损伤潜力的新药物。