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Cyanidin-3-Glucoside Reverses Ethanol-Induced Inhibition of Neurite Outgrowth: Role of Glycogen Synthase Kinase 3 Beta

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Abstract

Ethanol is a potent teratogen for the developing central nervous system (CNS), and fetal alcohol syndrome (FAS) is the most common nonhereditary cause of mental retardation. Ethanol disrupts neuronal differentiation and maturation. It is important to identify agents that provide neuroprotection against ethanol neurotoxicity. Using an in vitro neuronal model, mouse Neuro2a (N2a) neuroblastoma cells, we demonstrated that ethanol inhibited neurite outgrowth and the expression of neurofilament (NF) proteins. Glycogen synthase kinase 3β (GSK3β), a multifunctional serine/threonine kinase negatively regulated neurite outgrowth of N2a cells; inhibiting GSK3β activity by retinoic acid (RA) and lithium induced neurite outgrowth, while over-expression of a constitutively active S9A GSK3β mutant prevented neurite outgrowth. Ethanol inhibited neurite outgrowth by activating GSK3β through the dephosphorylation of GSK3β at serine 9. Cyanidin-3-glucoside (C3G), a member of the anthocyanin family rich in many edible berries and other pigmented fruits, enhanced neurite outgrowth by promoting p-GSK3β(Ser9). More importantly, C3G reversed ethanol-mediated activation of GSK3β and inhibition of neurite outgrowth as well as the expression of NF proteins. C3G also blocked ethanol-induced intracellular accumulation of reactive oxygen species (ROS). However, the antioxidant effect of C3G appeared minimally involved in its protection. Our study provides a potential avenue for preventing or ameliorating ethanol-induced damage to the developing CNS.

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Abbreviations

C3G:

Cyanidin-3-glucoside

FASD:

Fetal alcohol spectrum disorders

FAS:

Fetal alcohol syndrome

GSK3β:

Glycogen synthase kinase 3β

ROS:

Reactive oxygen species

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Acknowledgments

This research was supported by grants from the National Institutes of Health (AA015407), and the National Natural Science Foundation of China (30470544, 30471452, and 30570580). Dr. Z-J. Ke was also supported by the One Hundred Talents Program of the Chinese Academy of Sciences.

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Authors and Affiliations

  1. Department of Internal Medicine, University of Kentucky College of Medicine, 124C Combs Research Building, 800 Rose Street, Lexington, KY, 40536, USA

    Gang Chen, Kimberly A. Bower, Mei Xu & Jia Luo

  2. National Institute for Occupational Safety and Health, Morgantown, WV, 26505, USA

    Min Ding

  3. Graduate Center for Toxicology, University of Kentucky, 232 Bosomworth, Lexington, KY, 40536, USA

    Xianglin Shi

  4. Institute for Nutritional Sciences, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai, 200031, People’s Republic of China

    Zun-Ji Ke

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  1. Gang Chen
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  2. Kimberly A. Bower
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  3. Mei Xu
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  4. Min Ding
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  5. Xianglin Shi
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  6. Zun-Ji Ke
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  7. Jia Luo
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Correspondence to Jia Luo.

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Chen, G., Bower, K.A., Xu, M. et al. Cyanidin-3-Glucoside Reverses Ethanol-Induced Inhibition of Neurite Outgrowth: Role of Glycogen Synthase Kinase 3 Beta. Neurotox Res 15, 321–331 (2009). https://doi.org/10.1007/s12640-009-9036-y

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  • DOI: https://doi.org/10.1007/s12640-009-9036-y

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