Effect of dietary inducer dimethylfumarate on glutathione in cultured human retinal pigment epithelial cells

Kasey C. Nelson, Joanne L. Carlson, Melanie L. Newman, Paul Sternberg, Dean P. Jones, Terrance J. Kavanagh, Dolores Diaz, Jiyang Cai, Mei Wu

Research output: Contribution to journalArticle

72 Citations (Scopus)

Abstract

Purpose. To determine the effect of dimethylfumarate (DMF), an inducer of glutathione (GSH)-dependent detoxification, on intracellular GSH levels in cultured human retinal pigment epithelium (hRPE) cells, its mechanism of action, and its effect on hRPE cells subjected to oxidative injury. Methods. Established hRPE cell lines were treated with DMF and assayed by high- pressure liquid chromatography for intracellular and extracellular GSH levels. Quantification of γ-glutamylcysteine synthetase (GLCL) was determined through northern and western blot analyses, and activity was measured. Effects of pretreatment with DMF on GSH redox status of hRPE cells was determined. Sensitivity of hRPE cells to oxidative stress was determined using tert-butylhydroperoxide as the oxidative agent. Results. Dimethylfumarate caused a transient decrease followed by a significant increase in intracellular GSH. Glutathione increased maximally at 24 hours with 100 to 200 μM DMF. The initial decrease could be accounted for by the formation of a DMF-GSH conjugate. Dimethylfumarate treatment increased the steady state mRNA expression of the regulatory subunit of GLCL, but no increase was seen for the catalytic subunit. However, protein levels were increased for both, and the catalytic activity of GLCL was also increased. Whereas the initial decrease in GSH made hRPE cells more susceptible to oxidative damage, pretreatment with DMF under conditions that increased intracellular GSH protected hRPE cells against oxidative damage. Conclusions. These results suggest a means by which the antioxidant capability of hRPE may be augmented without direct antioxidant supplementation. Specifically, a dietary compound that conjugates with GSH can induce GSH synthesis, increase GSH concentration, and improve protection by GSH-dependent detoxification pathways in hRPE. However, the early depletion of GSH before stimulated synthesis necessitates caution in prevention strategies using dietary inducers.

Original languageEnglish (US)
Pages (from-to)1927-1935
Number of pages9
JournalInvestigative Ophthalmology and Visual Science
Volume40
Issue number9
StatePublished - Aug 1999
Externally publishedYes

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Retinal Pigments
Retinal Pigment Epithelium
Glutathione
Epithelial Cells
Antioxidants
Glutamate-Cysteine Ligase
tert-Butylhydroperoxide
Dimethyl Fumarate
Northern Blotting
Oxidation-Reduction
Catalytic Domain
Oxidative Stress
Western Blotting
High Pressure Liquid Chromatography
Cell Line
Messenger RNA
Wounds and Injuries

ASJC Scopus subject areas

  • Ophthalmology

Cite this

Nelson, K. C., Carlson, J. L., Newman, M. L., Sternberg, P., Jones, D. P., Kavanagh, T. J., ... Wu, M. (1999). Effect of dietary inducer dimethylfumarate on glutathione in cultured human retinal pigment epithelial cells. Investigative Ophthalmology and Visual Science, 40(9), 1927-1935.

Effect of dietary inducer dimethylfumarate on glutathione in cultured human retinal pigment epithelial cells. / Nelson, Kasey C.; Carlson, Joanne L.; Newman, Melanie L.; Sternberg, Paul; Jones, Dean P.; Kavanagh, Terrance J.; Diaz, Dolores; Cai, Jiyang; Wu, Mei.

In: Investigative Ophthalmology and Visual Science, Vol. 40, No. 9, 08.1999, p. 1927-1935.

Research output: Contribution to journalArticle

Nelson, KC, Carlson, JL, Newman, ML, Sternberg, P, Jones, DP, Kavanagh, TJ, Diaz, D, Cai, J & Wu, M 1999, 'Effect of dietary inducer dimethylfumarate on glutathione in cultured human retinal pigment epithelial cells', Investigative Ophthalmology and Visual Science, vol. 40, no. 9, pp. 1927-1935.
Nelson KC, Carlson JL, Newman ML, Sternberg P, Jones DP, Kavanagh TJ et al. Effect of dietary inducer dimethylfumarate on glutathione in cultured human retinal pigment epithelial cells. Investigative Ophthalmology and Visual Science. 1999 Aug;40(9):1927-1935.
Nelson, Kasey C. ; Carlson, Joanne L. ; Newman, Melanie L. ; Sternberg, Paul ; Jones, Dean P. ; Kavanagh, Terrance J. ; Diaz, Dolores ; Cai, Jiyang ; Wu, Mei. / Effect of dietary inducer dimethylfumarate on glutathione in cultured human retinal pigment epithelial cells. In: Investigative Ophthalmology and Visual Science. 1999 ; Vol. 40, No. 9. pp. 1927-1935.
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abstract = "Purpose. To determine the effect of dimethylfumarate (DMF), an inducer of glutathione (GSH)-dependent detoxification, on intracellular GSH levels in cultured human retinal pigment epithelium (hRPE) cells, its mechanism of action, and its effect on hRPE cells subjected to oxidative injury. Methods. Established hRPE cell lines were treated with DMF and assayed by high- pressure liquid chromatography for intracellular and extracellular GSH levels. Quantification of γ-glutamylcysteine synthetase (GLCL) was determined through northern and western blot analyses, and activity was measured. Effects of pretreatment with DMF on GSH redox status of hRPE cells was determined. Sensitivity of hRPE cells to oxidative stress was determined using tert-butylhydroperoxide as the oxidative agent. Results. Dimethylfumarate caused a transient decrease followed by a significant increase in intracellular GSH. Glutathione increased maximally at 24 hours with 100 to 200 μM DMF. The initial decrease could be accounted for by the formation of a DMF-GSH conjugate. Dimethylfumarate treatment increased the steady state mRNA expression of the regulatory subunit of GLCL, but no increase was seen for the catalytic subunit. However, protein levels were increased for both, and the catalytic activity of GLCL was also increased. Whereas the initial decrease in GSH made hRPE cells more susceptible to oxidative damage, pretreatment with DMF under conditions that increased intracellular GSH protected hRPE cells against oxidative damage. Conclusions. These results suggest a means by which the antioxidant capability of hRPE may be augmented without direct antioxidant supplementation. Specifically, a dietary compound that conjugates with GSH can induce GSH synthesis, increase GSH concentration, and improve protection by GSH-dependent detoxification pathways in hRPE. However, the early depletion of GSH before stimulated synthesis necessitates caution in prevention strategies using dietary inducers.",
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T1 - Effect of dietary inducer dimethylfumarate on glutathione in cultured human retinal pigment epithelial cells

AU - Nelson, Kasey C.

AU - Carlson, Joanne L.

AU - Newman, Melanie L.

AU - Sternberg, Paul

AU - Jones, Dean P.

AU - Kavanagh, Terrance J.

AU - Diaz, Dolores

AU - Cai, Jiyang

AU - Wu, Mei

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N2 - Purpose. To determine the effect of dimethylfumarate (DMF), an inducer of glutathione (GSH)-dependent detoxification, on intracellular GSH levels in cultured human retinal pigment epithelium (hRPE) cells, its mechanism of action, and its effect on hRPE cells subjected to oxidative injury. Methods. Established hRPE cell lines were treated with DMF and assayed by high- pressure liquid chromatography for intracellular and extracellular GSH levels. Quantification of γ-glutamylcysteine synthetase (GLCL) was determined through northern and western blot analyses, and activity was measured. Effects of pretreatment with DMF on GSH redox status of hRPE cells was determined. Sensitivity of hRPE cells to oxidative stress was determined using tert-butylhydroperoxide as the oxidative agent. Results. Dimethylfumarate caused a transient decrease followed by a significant increase in intracellular GSH. Glutathione increased maximally at 24 hours with 100 to 200 μM DMF. The initial decrease could be accounted for by the formation of a DMF-GSH conjugate. Dimethylfumarate treatment increased the steady state mRNA expression of the regulatory subunit of GLCL, but no increase was seen for the catalytic subunit. However, protein levels were increased for both, and the catalytic activity of GLCL was also increased. Whereas the initial decrease in GSH made hRPE cells more susceptible to oxidative damage, pretreatment with DMF under conditions that increased intracellular GSH protected hRPE cells against oxidative damage. Conclusions. These results suggest a means by which the antioxidant capability of hRPE may be augmented without direct antioxidant supplementation. Specifically, a dietary compound that conjugates with GSH can induce GSH synthesis, increase GSH concentration, and improve protection by GSH-dependent detoxification pathways in hRPE. However, the early depletion of GSH before stimulated synthesis necessitates caution in prevention strategies using dietary inducers.

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