Mechanisms of ammonia-induced cell death in rat cortical neurons: Roles of NMDA receptors and glutathione

被引:57
作者
Klejman, A
Wegrzynowicz, M
Szatmari, EM
Mioduszewska, B
Hetman, M
Albrecht, J
机构
[1] Nencki Inst Expt Biol, Dept Mol & Cellular Neurobiol, PL-02093 Warsaw, Poland
[2] Polish Acad Sci, Med Res Ctr, Dept Neurotoxicol, PL-02106 Warsaw, Poland
[3] Univ Louisville, Kentucky Spinal Cord Injury Res Ctr, Dept Neurol Surg, Louisville, KY 40202 USA
[4] Univ Louisville, Kentucky Spinal Cord Injury Res Ctr, Dept Pharmacol & Toxicol, Louisville, KY 40202 USA
关键词
ammonia; cortical neurons; apoptosis; Akt; excitotoxicity; glutathione;
D O I
10.1016/j.neuint.2005.04.006
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
The occurrence, nature and prevention of ammonia-induced cell death were assayed in cultured primary cortical neurons from newborn rats. Treatment with 1-10 mM ammonium chloride for 24 or 48 It, dose-dependently decreased neuronal survival (MTT assay) and GSH/ GSSG ratio in the cultures, whereas total GSH content was significantly reduced only with 10 mM ammonia. Treatment with a glutathione synthesis inhibitor, buthionyl sulfoximine (BSO) (10 mu M), decreased the GSH content and GSH/GSSG ratio to a degree similar to that of 10 mM ammonia, but it did not decrease cell survival in control cells. This indicates that glutathione depletion per se is not a cause of ammonia-induced neuronal death. However, ammonia-induced decrease of cell viability was attenuated by incubation with glutathione diethyl ester (GEE), which transiently increased the intracellular GSH level in both control and ammonia-treated cells. Neuronal survival in the presence of ammonia was partly improved by the NMDA receptor antagonists MK-801 and APV Morphological analysis revealed that ammonia treatment causes both apoptotic and non-apoptotic neuronal death, the former not being inhibited by MK-801. Apoptosis was the dominant type of cell death at 10 mM ammonia, as concluded both from morphologic examination and the absence of survival improvement in the presence of GABA + nipecotic acid or taurine, model anti-excitotoxic treatments of cortical neurons. The mechanism underlying apoptosis may include inhibition of a survival kinase, Akt, whose activatory phosphorylation at Ser473 is reduced in neurons treated with 10 mM, but not I mM ammonia. (c) 2005 Elsevier Ltd. All rights reserved.
引用
收藏
页码:51 / 57
页数:7
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