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Excitotoxicity Induced by Realgar in the Rat Hippocampus: the Involvement of Learning Memory Injury, Dysfunction of Glutamate Metabolism and NMDA Receptors

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Abstract

Realgar is a type of mineral drug containing arsenic. The nervous system toxicity of realgar has received extensive attention. However, the underlying mechanisms of realgar-induced neurotoxicity have not been clearly elucidated. To explore the mechanisms that contribute to realgar-induced neurotoxicity, weanling rats were exposed to realgar (0, 0.3, 0.9, 2.7 g/kg) for 6 weeks, and cognitive ability was tested using the Morris water maze (MWM) test and object recognition task (ORT). The levels of arsenic in the blood and hippocampus were monitored. The ultrastructures of hippocampal neurons were observed. The levels of glutamate (Glu) and glutamine (Gln) in the hippocampus and hippocampal CA1 region; the activities of glutamine synthetase (GS) and phosphate-activated glutaminase (PAG); the mRNA and protein expression of glutamate transporter 1 (GLT-1), glutamate/aspartate transporter (GLAST), and N-methyl-d-aspartate (NMDA) receptors; and the level of intracellular Ca2+ were also investigated. The results indicate that the rats developed deficiencies in cognitive ability after a 6-week exposure to realgar. The arsenic contained in realgar and the arsenic metabolites passed through the blood-brain barrier (BBB) and accumulated in the hippocampus, which resulted in the excessive accumulation of Glu in the extracellular space. The excessive accumulation of Glu in the extracellular space induced excitotoxicity, which was shown by enhanced GS and PAG activities, inhibition of GLT-1 mRNA and protein expression, alterations in NMDA receptor mRNA and protein expression, disturbance of intracellular Ca2+ homeostasis, and ultrastructural changes in hippocampal neurons. In conclusion, the findings from our study indicate that exposure to realgar induces excitotoxicity and that the mechanism by which this occurs may be associated with disturbances in Glu metabolism and transportation and alterations in NMDA receptor expression.

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Abbreviations

TCMs:

Traditional Chinese medicines

BBB:

Blood-brain barrier

MWM:

Morris water maze

ORT:

Object recognition task

Glu:

Glutamate

CNS:

Central nervous system

GLT-1:

Glutamate transporter 1

GLAST:

Glutamate/aspartate transporter

GS:

Glutamine synthetase

Gln:

Glutamine

PAG:

Phosphate-activated glutaminase

NMDA:

N-Methyl-d-aspartate

iAs:

Inorganic arsenic

MMA:

Methyl arsine

DMA:

Dimethyl arsine

ECL:

Enhanced chemiluminescence

CMC-Na:

Sodium carboxymethylcellulose

MD:

Microdialysis

NE:

Northeast

NW:

Northwest

SE:

Southeast

SW:

Southwest

AAS:

Atomic absorption spectrophotometry

TAs:

Total arsenic

HPLC:

High-performance liquid chromatography

PVDF:

Polyvinylidene difluoride

TBS:

Tris-buffered saline

PBS:

Phosphate-buffered saline

NOAEL:

No observed adverse effect level

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Acknowledgments

The work is financially supported by the National Natural Science Foundation of China (81073144), the China Postdoctoral Science Foundation on the 51th grant program (2012M510848), and the Higher Education Department of Liaoning Province (L2010708).

Conflict of Interest

The authors declare that they have no conflicts of interest.

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Authors and Affiliations

  1. Department of Health Laboratory Technology, School of Public Health, China Medical University, No. 92 of Bei Er Road, Shenyang, 110001, People’s Republic of China

    Tao-guang Huo, Wei-kai Li, Ying-hua Zhang, Jie Yuan, Lan-yue Gao, Yuan Yuan, Hui-lei Yang & Hong Jiang

  2. Liaoning Provincial Key Laboratory of Arsenic Biological Effect and Poisoning, China Medical University, No. 92 of Bei Er Road, Shenyang, 110001, People’s Republic of China

    Tao-guang Huo, Hong Jiang & Gui-fan Sun

  3. Department of Occupational and Environmental Health, School of Public Health, China Medical University, No. 92 of Bei Er Road, Shenyang, 110001, People’s Republic of China

    Gui-fan Sun

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  1. Tao-guang Huo
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  2. Wei-kai Li
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  4. Jie Yuan
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  8. Hong Jiang
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Correspondence to Hong Jiang.

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Huo, Tg., Li, Wk., Zhang, Yh. et al. Excitotoxicity Induced by Realgar in the Rat Hippocampus: the Involvement of Learning Memory Injury, Dysfunction of Glutamate Metabolism and NMDA Receptors. Mol Neurobiol 51, 980–994 (2015). https://doi.org/10.1007/s12035-014-8753-2

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  • DOI: https://doi.org/10.1007/s12035-014-8753-2

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