Involvement of Oxidative Stress on the Impairment of Energy Metabolism Induced by Aβ Peptides on PC12 Cells: Protection by Antioxidants

doi:10.1006/nbdi.1999.0241

Neurobiology of Disease

Volume 6, Issue 3, June 1999, Pages 209-219

https://doi.org/10.1006/nbdi.1999.0241 Get rights and content

Abstract

Alzheimer's disease is widely held to be associated with oxidative stress due, in part, to the membrane action of amyloid β-peptide (Aβ) aggregates. In this study, the involvement of oxidative stress on Aβ-induced energy metabolism dysfunction was evaluated on PC12 cells. It was shown that Aβ peptides (Aβ25–35 and Aβ1–40) induce a concentration-dependent accumulation of reactive oxygen species (ROS), decrease the cellular redox activity, and lead to the depletion of ATP levels. The observed inhibition by Aβ of mitochondrial function and of glycolysis is blocked by the antioxidants vitamin E, idebenone, and GSH ethyl ester. Taken together, these data suggest that exposure of PC12 cells to Aβ results in an impairment of energy metabolism, leading to a deficit in ATP levels and to the compromise of cellular viability. Furthermore, the generation of ROS seems to be a crucial event responsible for the energetic metabolic dysfunction induced by Aβ.

References (63)

C. Behl et al.
Vitamin E protects nerve cells from amyloid β protein toxicity
Biochem. Biophys. Res. Commun.
(1992)
C. Behl et al.
Hydrogen peroxide mediates amyloid β protein toxicity
Cell
(1994)
G. Benzi et al.
Age- and peroxidative stress-related modifications of the cerebral enzymatic activities linked to mitochondria and the glutathione system
Free Radical Biol. Med.
(1995)
J.P.V. Bolaños et al.
Nitric oxide-mediated mitochondrial damage: A possible neuroprotective role for glutathione
Free Radical Biol. Med.
(1996)
A.C. Bowling et al.
Bioenergetic and oxidative stress in neurodegenerative diseases
Life Sci.
(1995)
D.L. Brautigan et al.
Mitochondrial cytochrome c: Preparation and activity of native and chemically modified cytochrome c
Methods Enzymol.
(1978)
D.A. Butterfield et al.
β-Amyloid peptide free radical fragments initiate synaptosomal lipoperoxidation in a sequence-specific fashion: Implications to Alzheimer's disease
Biochem. Biophys. Res. Commun.
(1994)
R. Cathcart et al.
Detection of picomole levels of hydroperoxides using a fluorescent dichlorofluorescein assay
Anal. Biochem.
(1983)
J.J. Chen et al.
Alterations in mitochondrial membrane fluidity by lipid peroxidation products
Free Radical Biol. Med.
(1994)
H. Esterbauer et al.
Chemistry and biochemistry of 4-hydroxynonenal, malonaldehyde and related aldehydes
Free Radical Biol. Med.
(1991)

Y. Goodman et al.

Secreted forms of β-amyloid precursor protein protect hippocampal neurons against amyloid β-peptide-induced oxidative injury

Exp. Neurol.

(1994)

J.T. Greenamyre et al.

Excitatory amino acids and Alzheimer's disease

Neurobiol. Aging

(1989)

D.P. Hartley et al.

The hepatocellular metabolism of 4-hydroxynonenal by alcohol dehydrogenase, aldehyde dehydrogenase and glutathione S-transferase

Arch. Biochem. Biophys.

(1995)

J.N. Keller et al.

4-Hydroxynonenal, an aldehydic product of membrane lipid peroxidation, impairs glutamate transport and mitochondrial function in synaptosomes

Neuroscience

(1997)

A.M. Kennedy et al.

Deficits in cerebral glucose metabolism demonstrated by positron emission tomography in individuals at risk of familiar Alzheimer's disease

Neurosci. Lett.

(1995)

W.R. Markesbery et al.

Four-hydroxynonenal, a product of lipid peroxidation, is increased in the brain in Alzheimer's disease

Neurobiol. Aging

(1998)

M.P. Mattson et al.

β-Amyloid precursor protein metabolites and loss of neuronal Ca²⁺ homeostasis in Alzheimer's disease

Trends Neurosci.

(1993)

M.P. Mattson et al.

Calcium, free radicals, and excitotoxic neuronal death in primary cell culture

Methods Cell Biol.

(1995)

I. Miwa et al.

4-Hydroxy-2-nonenal hardly affects glycolysis

Free Radical Biol. Med.

(1997)

T. Mosmann

Rapid colorimetric assay for cellular growth and survival

J. Immunol. Methods

(1983)

J.J. Sedmak et al.

A rapid sensitive and versatile assay for protein using Coomassie blue G 250

Ann. Biochem.

(1977)

M.A. Smith et al.

Radical ageing in Alzheimer's disease

Trends Neurosci.

(1995)

V. Stocchi et al.

Simultaneous extraction and reverse-phase high-performance liquid chromatography determination of adenine and pyridine nucleotides in human red blood cells

Anal. Biochem.

(1985)

H.D. Tisdale

Preparation and properties of succinic-cytochrome c reductase (complex II and III)

Methods Enzymol.

(1967)

D.A. Bass et al.

Flow cytometric studies of oxidative product formation by neutrophils: A graded response to membrane stimulation

J. Immunol.

(1983)

C. Behl et al.

Protection against oxidative stress-induced neuronal cell death—A novel role for RU486

Eur. J. Neurosci.

(1997)

E.M. Blanc et al.

4-Hydroxynonenal, a lipid peroxidation product, impairs glutamate transport in cortical astrocytes

Glia

(1998)

E.M. Blanc et al.

Amyloid β-peptide induces cell monolayer albumin permeability, impairs glucose transport, and induces apoptosis in vascular endothelial cell

J. Neurochem.

(1997)

A.J. Bruce et al.

β-Amyloid toxicity in organotypic cultures: Protection by EUK-8, a synthetic catalytic free radical scavenger

Proc. Natl. Acad. Sci. USA

(1996)

S. Desagher et al.

Pyruvate protects neurons against hydrogen peroxide-induced toxicity

J. Neurosci.

(1997)

L.L. Dyrks et al.

Amyloidogenicity of βA4 and βA4-bearing amyloid protein precursor fragments by metal-catalyzed oxidation

J. Biol. Chem.

(1992)

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Regular ArticleInvolvement of Oxidative Stress on the Impairment of Energy Metabolism Induced by Aβ Peptides on PC12 Cells: Protection by Antioxidants

Abstract

Biochem. Biophys. Res. Commun.

Cell

Free Radical Biol. Med.

Free Radical Biol. Med.

Life Sci.

Methods Enzymol.

Biochem. Biophys. Res. Commun.

Anal. Biochem.

Free Radical Biol. Med.

Free Radical Biol. Med.

Exp. Neurol.

Neurobiol. Aging

Arch. Biochem. Biophys.

Neuroscience

Neurosci. Lett.

Neurobiol. Aging

Trends Neurosci.

Methods Cell Biol.

Free Radical Biol. Med.

J. Immunol. Methods

Ann. Biochem.

Trends Neurosci.

Anal. Biochem.

Methods Enzymol.

Flow cytometric studies of oxidative product formation by neutrophils: A graded response to membrane stimulation

J. Immunol.

Protection against oxidative stress-induced neuronal cell death—A novel role for RU486

Eur. J. Neurosci.

4-Hydroxynonenal, a lipid peroxidation product, impairs glutamate transport in cortical astrocytes

Glia

Amyloid β-peptide induces cell monolayer albumin permeability, impairs glucose transport, and induces apoptosis in vascular endothelial cell

J. Neurochem.

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Proc. Natl. Acad. Sci. USA

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J. Neurosci.

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J. Biol. Chem.

Regular Article
Involvement of Oxidative Stress on the Impairment of Energy Metabolism Induced by Aβ Peptides on PC12 Cells: Protection by Antioxidants