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Melatonin Antagonizes Mn-Induced Oxidative Injury Through the Activation of Keap1–Nrf2–ARE Signaling Pathway in the Striatum of Mice

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Abstract

Excessive manganese (Mn) exposure can lead to oxidative injury. Nuclear factor erythroid 2-related factor 2 (Nrf2) exerts an antioxidant response toward various environmental toxicants in the brain. However, the role of Nrf2 against Mn-induced oxidative injury remains largely unexplored. This study investigated the role of melatonin (MLT), an agent that was recently shown to induce the activation of the Kelch-like ECH-associated protein 1 (Keap1)–Nrf2–antioxidant response elements (ARE) pathway against manganism. Mice were randomly divided into six groups, including control, 12.5, 25, 50 mg/kg MnCl2, MLT control, and MLT + 50 mg/kg MnCl2. The following were determined: behavioral activity; pathological changes; immunofluorescence staining of Neuronal Nuclei and glial fibrillary acidic protein; cell apoptosis; the levels of reactive oxygen species (ROS), malondialdehyde (MDA), and glutathione (GSH); the immunohistochemical expression; and the protein levels of Nrf2, Keap1, and downstream enzymes. Mn-induced motor disorders, pathological damage, neuron degeneration, astrocytes activation, apoptosis, ROS and MDA generation, and GSH depletion. Nrf2, keap1, heme oxygenase-1 and NAD(P)H dehydrogenase, and quinone 1 showed a biphasic expression trend, which was most evidenced in the 12.5 mg/kg MnCl2 group. Changes in γ-glutamylcysteine synthetase, glutathione peroxidase 1, glutathionine S-transferase, glutathione reductase, and superoxide dismutase decreased in a concentration-dependent manner as a result of Mn exposure. MLT antagonized oxidative injury through the activation of the Keap1–Nrf2–ARE signaling pathway. In conclusion, disturbance of the Keap1–Nrf2–ARE signaling pathway partly caused oxidative injury. MLT can activate Nrf2 and its downstream enzymes and reverse Mn-induced oxidative injury.

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Abbreviations

Mn:

Manganese

MnO2 :

Manganese dioxide

PD:

Parkinson’s disease

MMT:

Methylcyclopentadienyl manganese tricarbonyl

HD:

Huntington’s disease

AD:

Alzheimer’s disease

ALS:

Amyotrophic lateral sclerosis

BBB:

Blood–brain barrier

ROS:

Reactive oxygen species

ATP:

Adenosine triphosphate

Nrf2:

Nuclear factor erythroid 2-related factor 2

Keap1:

Kelch-like ECH-associated protein 1

ARE:

Antioxidant response elements

HO-1:

Heme oxygenase-1

NQO1:

NAD(P)H dehydrogenase, quinone 1

SOD:

Superoxide dismutase

γ-GCS:

γ-Glutamylcysteine synthetase

GPx-1:

Glutathione peroxidase 1

GR:

Glutathione reductase

GST:

Glutathionine S-transferase

MLT:

N-Acetyl-5-methoxytryptamine

–HO· :

Hydroxyl radical

NO· :

Nitric oxide

O −·2 :

Superoxide anion

GSH:

Glutathione

MDA:

Malondialdehyde

MnCl2·4H2O:

Manganese chloride

DCFH-DA:

2,7-Dichlorofluorescin-diacetate

SABC:

Strept avidin–biotin complex

DAB:

Diaminobenzidine

HE:

Hematoxylin and eosin

BSA:

Bovine serum albumin

DCF:

Dichlorofluorescein

PBS:

Phosphate-buffered saline

TCA:

Trichloroacetic acid

TBA:

Thiobarbituric acid

TEP:

Tetraethoxypropane

DTNB:

1,2-Dithio-bis-nitrobenzoic acid

IOD:

Integral optical density

PVDF:

Polyvinylidene difluoride

ANOVA:

One-way analysis of variance

bZIP:

Basic region leucine zipper

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Acknowledgments

The Grants from the National Natural Science Foundation of China (No. 81302406) and the Specialized Research Fund of New Teachers for the Doctoral Program of Higher Education of China (20112104120017) supported this work financially.

Conflict of interest

The authors declare that there are no conflicts of interest.

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Authors and Affiliations

  1. Department of Environmental Health, School of Public Health, China Medical University, Shenyang, 110001, Liaoning, People’s Republic of China

    Yu Deng, Jiayu Zhu, Bin Xu, Congcong Jiao, Yuehui Li, Wei Liu & Zhaofa Xu

  2. Department of School Health Supervision, Institute of Shenyang Health Inspection, Shenyang, 110001, Liaoning, People’s Republic of China

    Chao Mi

  3. Department of Pharmacy, First Affiliated Hospital of China Medical University, Shenyang, People’s Republic of China

    Donghui Xu

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  1. Yu Deng
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  2. Jiayu Zhu
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Correspondence to Yu Deng.

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Deng, Y., Zhu, J., Mi, C. et al. Melatonin Antagonizes Mn-Induced Oxidative Injury Through the Activation of Keap1–Nrf2–ARE Signaling Pathway in the Striatum of Mice. Neurotox Res 27, 156–171 (2015). https://doi.org/10.1007/s12640-014-9489-5

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