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Dopamine as a Potent Inducer of Cellular Glutathione and NAD(P)H:Quinone Oxidoreductase 1 in PC12 Neuronal Cells: A Potential Adaptive Mechanism for Dopaminergic Neuroprotection

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Abstract

Dopamine auto-oxidation and the consequent formation of reactive oxygen species and electrophilic quinone molecules have been implicated in dopaminergic neuronal cell death in Parkinson’s disease. We reported here that in PC12 dopaminergic neuronal cells dopamine at noncytotoxic concentrations (50–150 μM) potently induced cellular glutathione (GSH) and the phase 2 enzyme NAD(P)H:quinone oxidoreductase 1 (NQO1), two critical cellular defenses in detoxification of ROS and electrophilic quinone molecules. Incubation of PC12 cells with dopamine also led to a marked increase in the mRNA levels for γ-glutamylcysteine ligase catalytic subunit (GCLC) and NQO1. In addition, treatment of PC12 cells with dopamine resulted in a significant elevation of GSH content in the mitochondrial compartment. To determine whether treatment with dopamine at noncytotoxic concentrations, which upregulated the cellular defenses could protect the neuronal cells against subsequent lethal oxidative and electrophilic injury, PC12 cells were pretreated with dopamine (150 μM) for 24 h and then exposed to various cytotoxic concentrations of dopamine or 6-hydroxydopamine (6-OHDA). We found that pretreatment of PC12 cells with dopamine at a noncytotoxic concentration led to a remarkable protection against cytotoxicity caused by dopamine or 6-OHDA at lethal concentrations, as detected by 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium reduction assay. In view of the critical roles of GSH and NQO1 in protecting against dopaminergic neuron degeneration, the above findings implicate that upregulation of both GSH and NQO1 by dopamine at noncytotoxic concentrations may serve as an important adaptive mechanism for dopaminergic neuroprotection.

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Abbreviations

CDNB:

1-Chloro-2,4-dinitrobenzene

DCIP:

2,6-Dichloroindophenol

DMEM:

Dulbecco’s modified Eagle’s medium

FBS:

Fetal bovine serum

GAPD:

Hglyceraldehyde-3-phosphate dehydrogenase

GCLC:

γ-Glutamylcysteine ligase catalytic subunit

GPx:

Glutathione peroxidase

GR:

Glutathione reductase

GSH:

Reduced glutathione

GSSG:

Oxidized form of glutathione

GST:

Glutathione S-transferase

MTT:

3-[4,5-Dimethylthiazol-2-yl]-2,5-diphenyltetrazolium

NQO1:

NAD(P)H:quinone oxidoreductase 1

Nrf2:

Nuclear factor E2-related factor 2

6-OHDA:

6-Hydroxydopamine

PD:

Parkinson’s disease

PBS:

Phosphate buffered saline

ROS:

Reactive oxygen species

SOD:

Superoxide dismutase

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Acknowledgments

This work was supported in part by NIH grant HL71190 (Y.L.), and a grant from Harvey Peters Research Foundation (H.P.M.).

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Authors and Affiliations

  1. Division of Biomedical Sciences, Edward Via Virginia College of Osteopathic Medicine, Virginia Tech Corporate Research Center, 2265 Kraft Drive, Blacksburg, VA, 24060, USA

    Zhenquan Jia, Hong Zhu, Bhaba R. Misra, Yunbo Li & Hara P. Misra

  2. Department of Biomedical Sciences and Pathobiology, Virginia-Maryland Regional College of Veterinary Medicine, Virginia Tech, Blacksburg, VA, 24061, USA

    Yunbo Li & Hara P. Misra

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  1. Zhenquan Jia
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  2. Hong Zhu
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  3. Bhaba R. Misra
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  4. Yunbo Li
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Correspondence to Yunbo Li or Hara P. Misra.

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Jia, Z., Zhu, H., Misra, B.R. et al. Dopamine as a Potent Inducer of Cellular Glutathione and NAD(P)H:Quinone Oxidoreductase 1 in PC12 Neuronal Cells: A Potential Adaptive Mechanism for Dopaminergic Neuroprotection. Neurochem Res 33, 2197–2205 (2008). https://doi.org/10.1007/s11064-008-9670-4

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  • DOI: https://doi.org/10.1007/s11064-008-9670-4

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