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Mitochondrion-Mediated Apoptosis Induced by Acrylamide is Regulated by a Balance Between Nrf2 Antioxidant and MAPK Signaling Pathways in PC12 Cells

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Abstract

Acrylamide (ACR) is a potent toxin that affects the human nervous system. However, the underlying mechanism of ACR neurotoxicity remains poorly understood. In the present study, we investigated whether ACR induces mitochondrion-dependent apoptosis and the involved signaling pathways in PC12 cells. ACR exposure activated the mitochondrial apoptotic pathway in PC12 cells and triggered the up-regulation of Bax/Bcl-2 ratio, excessive release of cytochrome c, cleavage of capase-9 and caspase-3, depolarization of the mitochondrial membrane, structural damages to the mitochondria, and compaction of nuclear heterochromatin. ACR-induced oxidative stress was also observed based on distinct increase in cellular reactive oxygen species (ROS) and malondialdehyde (MDA), and significant decrease in glutathione (GSH). Mitogen-activated protein kinases (MAPK) signaling including extracellular signal-regulated protein kinases (ERK), c-Jun N-terminal kinases (JNK), and p38 were phosphorylated by ROS overproduction in PC12 cells in a time-and dose-dependent manner. ACR promoted the translocation of nuclear factor E2-related factor 2 (Nrf2) from the cytosol to the nucleus, thereby enhancing the expression of downstream γ-glutamyl-cysteine synthetase (γ-GCS). The regulation of Nrf2 activation by MAPK pathways was confirmed by the blockade of MAPK pathways. The suppression of JNK and p38 pathways showed a protective effect on ACR-induced mitochondrial dysfunction and apoptosis. Nrf2 knockdown further enhanced MDA production and reduced GSH generation induced by ACR. These results suggest that MAPK and Nrf2 signaling pathways contribute to mitochondrion-mediated apoptosis induced by ACR in PC12 cells.

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Abbreviations

ACR:

Acrylamide

Nrf2:

Nuclear transcription factor E2-related factor 2

MAPK:

Mitogen activated protein kinase

ERK:

Extracellular signal regulated protein kinase

JNK:

c-Jun N-terminal kinase

ROS:

Reactive oxygen species

MDA:

Malondialdehvde

GSH:

Glutathione

γ-GCS:

γ-Glutamyl-cysteine synthetase

MMP:

Mitochondrial membrane potential

DMSO:

Dimethylsulfoxide

FBS:

Fetal bovine serum

PI:

Propidium iodide

MTT:

3–4, 5-Dimethylthiazol-2-yl-2, 5-diphenyltetrazoliumbromide

JC-1:

5–5, 6–6-Tetrachloro-1, 1, 3, 3-tetra-ethylbenzimidazolcarbocya-nine iodide

DAPI:

4′,6-Diamidino-2-phenylindole

DCFH-DA:

2, 7-Dichlorofluorescein diacetate

TBA:

Thiobarbituric acid;

BSA:

Bull serum albumin

PBS:

Phosphate buffer solution

TEM:

Transmission electron microscopy

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Acknowledgments

We appreciate the contribution of all the members participating in this study. The work was financially supported by the National Natural Science Foundation of China (No. 81373042).

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Authors and Affiliations

  1. Department of Health Toxicology, MOE Key Lab of Environment and Health, School of Public Health, Tongji Medical College, Huazhong University of Science and Technology, 13 Hangkong-Road, Wuhan, Hubei, 430030, People’s Republic of China

    Xiaoqi Pan, Dandan Yan, Dun Wang, Xu Wu, Wanyun Zhao, Qing Lu & Hong Yan

  2. School of Public Health, Chengdu University of Traditional Chinese Medicine, Chengdu, 611130, China

    Xiaoqi Pan

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  1. Xiaoqi Pan
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  2. Dandan Yan
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Correspondence to Hong Yan.

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Pan, X., Yan, D., Wang, D. et al. Mitochondrion-Mediated Apoptosis Induced by Acrylamide is Regulated by a Balance Between Nrf2 Antioxidant and MAPK Signaling Pathways in PC12 Cells. Mol Neurobiol 54, 4781–4794 (2017). https://doi.org/10.1007/s12035-016-0021-1

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  • DOI: https://doi.org/10.1007/s12035-016-0021-1

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