Nitric oxide-mediated mitochondrial damage: A potential neuroprotective role for glutathione
Abstract
In this study we have investigated the mechanisms leading to mitochondrial damage in cultured neurons following sustained exposure to nitric oxide. Thus, the effects upon neuronal mitochondrial respiratory chain complex activity and reduced glutathione concentration following exposure to either the nitric oxide donor, S-nitroso-N-acetylpenicillamine, or to nitric oxide releasing astrocytes were assessed. Incubation with S-nitroso-N-acetylpenicillamine (1 mM) for 24 h decreased neuronal glutathione concentration by 57%, and this effect was accompanied by a marked decrease of complex I (43%), complex II-III (63%), and complex IV (41%) activities. Incubation of neurons with the glutathione synthesis inhibitor, L-buthionine-[S,R]-sulfoximine caused a major depletion of neuronal glutathione (93%), an effect that was accompanied by a marked loss of complex II-III (60%) and complex IV (41%) activities, although complex I activity was only mildly decreased (34%). In an attempt to approach a more physiological situation, we studied the effects upon glutathione status and mitochondrial respiratory chain activity of neurons incubated in coculture with nitric oxide releasing astrocytes. Astrocytes were activated by incubation with lipopolysaccharide/interferon-gamma for 18 h, thereby inducing nitric oxide synthase and, hence, a continuous release of nitric oxide. Coincubation for 24 h of activated astrocytes with neurons caused a limited loss of complex IV activity and had no effect on the activities of complexes I or II-III. However, neurons exposed to astrocytes had a 1.7-fold fold increase in glutathione concentration compared to neurons cultured alone. Under these coculture conditions, the neuronal ATP concentration was modestly reduced (14%). This loss of ATP was prevented by the nitric oxide synthase inhibitor, NG-monomethyl-L-arginine. These results suggest that the neuronal mitochondrial respiratory chain is damaged by sustained exposure to nitric oxide and that reduced glutathione may be an important defence against such damage.
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In co-cultures, the presence of astrocytes increases the GSH content in the cocultured neurons.
“Several groups have shown that in co‐cultures, the presence of astrocytes increases the GSH content in the cocultured neurons (Bolaños et al. 1996; Diaz‐Hernandez et al. 2005; Dringen, Pfeiffer, et al. 1999; Gegg et al. 2005; Pizzurro et al. 2014; Rathinam et al. 2012), demonstrating that the presence of astrocytes has a stimulating positive effect on the GSH content of neurons.”
From Glutathione Metabolism of the Brain—The Role of Astrocytes (2025) · cites this paper as Bolaños et al. 1996
A high astrocytic GSH content is essential to provide neighboring neurons with amino acid precursors for neuronal GSH synthesis.
“A high astrocytic GSH content of astrocytes is essential to provide neighboring neurons with amino acid precursors for neuronal GSH synthesis.”
From Glutathione Metabolism of the Brain—The Role of Astrocytes (2025) · cites this paper as Bolaños et al. 1996
The decrease in maximum voluntary contraction for ISGE may result from excess nitric oxide disrupting mitochondrial oxidative capacity.
“may result from the potential for excess nitric oxide to disrupt mitochondrial oxidative capacity (Bolanos et al. 1996; Clementi et al. 1998)”
From Oral nitrate and citrulline decrease blood pressure and increase vascular conductance in young adults: a potential therapy for heart failure (2016) · cites this paper as Bolanos et al. 1996
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- Authors
- Bolaños, J. P., Heales, S. J. R., Peuchen, S., Barker, J. E., Land, J. M., & Clark, J. B.
- Affiliations
- UCL Queen Square Institute of Neurology (GB)
- University of London (GB)
- Corresponding author
- Bolaños, Juan P.
- Venue
- Free Radical Biology and Medicine
- Publisher
- Elsevier BV
- Published
- 1996
- Volume
- 21
- Issue
- 7
- Pages
- 995-1001
- ISSN
- 0891-5849 (ISSN-L)
- OpenAlex
- W2093283930
- Cited by
- 263
- References
- 48
- Citations by year
- 2026: 2 · 2025: 6 · 2024: 5 · 2023: 2 · 2022: 2 · 2021: 4 · 2020: 9 · 2019: 3 · 2018: 6 · 2017: 2 · 2016: 6 · 2015: 6 · 2014: 9 · 2013: 4 · 2012: 8
- FWCI
- 4.64
- Type
- journal article
- Language
- en
- Open access
- closed
- Locations
- Free Radical Biology and Medicine (publishedVersion)
- PubMed (publishedVersion)
- UCL Discovery (University College London) (submittedVersion)
- Funders
- Wellcome Trust
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Mitochondrial Function and Pathology
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Redox biology and oxidative stress
- Life Sciences › Neuroscience › Neurology › Neurological Disorders and Treatments
- Keywords
- nitric oxide, mitochondrial damage, glutathione, neuronal mitochondria, respiratory chain complexes, S-nitroso-N-acetylpenicillamine, astrocytes, neuroprotection, glutathione synthesis, L-buthionine sulfoximine, nitric oxide synthase, L-NMMA, ATP depletion, astrocyte-neuron co-culture
- MeSH terms
- Animals, Astrocytes, Cell Survival, Cells, Cultured, Citrate (si)-Synthase, Electron Transport, Energy Metabolism, Female, Glutathione, Mitochondria, Neurons, Nitric Oxide, Penicillamine, Rats, Wistar, Coculture Techniques, Nitric Oxide Synthase, S-Nitroso-N-Acetylpenicillamine, Rats
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