Necrostatin-1 protects against glutamate-induced glutathione depletion and caspase-independent cell death in HT-22 cells

• Published in: Journal of neurochemistry 2007-12, Vol.103 (5), p.2004-2014
• Main Authors: Xu, Xingshun, Chua, Chu C, Kong, Jiming, Kostrzewa, Richard M, Kumaraguru, Udayasankar, Hamdy, Ronald C, Chua, Balvin H.L
• Format: Article
• Language: English
• Subjects: Activating Transcription Factors - metabolism; Animals; Apoptosis; apoptosis-inducing factor; Bcl-2/adenovirus E1B 19 kDa-interacting protein 3; Biological and medical sciences; Blood Proteins - metabolism; Caspases - metabolism; Cell Death - drug effects; Cell Line, Transformed; Cell Survival - drug effects; Cells; Chemistry; Degenerative and inherited degenerative diseases of the nervous system. Leukodystrophies. Prion diseases; Drug Interactions; Enzymes; glutamate toxicity; Glutamic Acid - pharmacology; Glutathione - metabolism; HT-22 cells; Imidazoles - pharmacology; Indoles - pharmacology; Medical sciences; Membrane Proteins - metabolism; Mice; Mitochondria - drug effects; Mitochondria - metabolism; Mitochondrial Proteins - metabolism; necroptosis; necrostatin-1; Neurological disorders; Neurology; Neurons - drug effects; Neurosciences; Neurotransmitters; Protein Transport - drug effects; Reactive Oxygen Species - metabolism; Signal Transduction - drug effects; Vascular diseases and vascular malformations of the nervous system
• ISSN: 0022-3042; 1471-4159
• DOI: 10.1111/j.1471-4159.2007.04884.x

Glutamate, a major excitatory neurotransmitter in the CNS, plays a critical role in neurological disorders such as stroke and Parkinson's disease. Recent studies have suggested that glutamate excess can result in a form of cell death called glutamate-induced oxytosis. In this study, we explore the protective effects of necrostatin-1 (Nec-1), an inhibitor of necroptosis, on glutamate-induced oxytosis. We show that Nec-1 inhibits glutamate-induced oxytosis in HT-22 cells through a mechanism that involves an increase in cellular glutathione (GSH) levels as well as a reduction in reactive oxygen species production. However, Nec-1 had no protective effect on free radical-induced cell death caused by hydrogen peroxide or menadione, which suggests that Nec-1 has no antioxidant effects. Interestingly, the protective effect of Nec-1 was still observed when cellular GSH was depleted by buthionine sulfoximine, a specific and irreversible inhibitor of glutamylcysteine synthetase. Our study further demonstrates that Nec-1 significantly blocks the nuclear translocation of apoptosis-inducing factor (a marker of caspase-independent programmed cell death) and inhibits the integration of Bcl-2/adenovirus E1B 19 kDa-interacting protein 3 (a pro-death member of the Bcl-2 family) into the mitochondrial membrane. Taken together, these results demonstrate for the first time that Nec-1 prevents glutamate-induced oxytosis in HT-22 cells through GSH related as well as apoptosis-inducing factor and Bcl-2/adenovirus E1B 19 kDa-interacting protein 3-related pathways.