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Metabolic and Antioxidant System Alterations in an Astrocytoma Cell Line Challenged with Mitochondrial DNA Deletion

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Abstract

Oxidative stress can induce mitochondrial dysfunction, mitochondrial DNA (mtDNA) depletion, and neurodegeneration, although the underlying mechanisms are poorly understood. The major mitochondrial antioxidant system that protects cells consists of manganese superoxide dismutase (MnSOD), glutathione peroxidase (GPx) and glutathione (GSH). To investigate the putative adaptive changes in antioxidant enzyme protein expression and targeting to mitochondria as mtDNA depletion occurs, we progressively depleted U87 astrocytoma cells of mtDNA by chronic treatment with ethidium bromide (EB, 50 ng/ml). Cellular MnSOD protein expression was markedly increased in a time-related manner while that of GPx showed time-related decreases. The mtDNA depletion also altered targeting or subcellular distribution of GPx, suggesting the importance of intact mtDNA in mitochondrial genome-nuclear genome signaling/communication. Cellular NADP+-ICDH activity also showed marked, time-related increases while their GSH content decreased. Thus, our findings suggest that interventions to elevate MnSOD, GPx, NADP+-ICDH, and GSH levels may protect brain cells from oxidative stress.

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Abbreviations

AD:

Alzheimer’s disease

AZT:

Azidothymidine

BCA:

Bicinchoninic acid

BSA:

Bovine serum albumin

DMEM:

Dulbecco’s modified essential medium

ETC:

Electron transport chain

EB:

Ethidium bromide

GPx:

Glutathione peroxidase

GSH:

Glutathione

HEPES:

N-2 hydroxyethylpiperazine-N`-2 ethane sulfonic acid

ICDH:

Isocitrate dehydrogenase

LDH:

Lactate dehydrogenase

MnSOD:

Manganese superoxide dismutase

mtDNA:

Mitochondrial DNA

NADP+ :

Nicotinamide adenine dinucleotide phosphate

nDNA:

Nuclear DNA

PVDF:

Polyvinylidene fluoride

PD:

Parkinson’s disease

ROS:

Reactive oxygen species

SDS:

Sodium dodecyl sulfate

TCA:

Tricarboxylic acid

U87:

A human astrocytoma cell line

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Acknowledgements

Our studies were supported by a grant from Idaho Biomedical Research Infrastructure Network (NIH NCRR BRINIP20RR016454), and a small project grant from the Mountain States Tumor and Medical Research Institute.

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Authors and Affiliations

  1. Department of Pharmaceutical Sciences, College of Pharmacy and Biomedical Research Institute, Idaho State University, Pocatello, ID, 83209, USA

    Alfred Orina Isaac, Vikas V. Dukhande & James C. K. Lai

  2. Mountain States Tumor and Medical Research Institute, Boise, ID, 83712, USA

    James C. K. Lai

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  1. Alfred Orina Isaac
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  2. Vikas V. Dukhande
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Correspondence to James C. K. Lai.

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Isaac, A.O., Dukhande, V.V. & Lai, J.C.K. Metabolic and Antioxidant System Alterations in an Astrocytoma Cell Line Challenged with Mitochondrial DNA Deletion. Neurochem Res 32, 1906–1918 (2007). https://doi.org/10.1007/s11064-007-9380-3

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