Vitamin C protects against ethanol and PTZ-induced apoptotic neurodegeneration in prenatal rat hippocampal neurons

Synapse. 2011 Jul;65(7):562-71. doi: 10.1002/syn.20875. Epub 2010 Dec 3.


Exposure to alcohol during brain development may cause a neurological syndrome called fetal alcohol syndrome, characterized by pre- and postnatal growth deficiencies, craniofacial anomalies, and evidence of CNS dysfunction. The objective of this study was to evaluate pentylenetetrazol (PTZ) and ethanol effects on Bax, Bcl-2 expression, which further induced activation of caspase-3, release of cytochrome-c from mitochondria, and to observe the protective effects of vitamin C (vit-C) against PTZ and ethanol-induced apoptotic neurodegeneration in primary-cultured neuronal cells at gestational day 17.5. Apoptotic neurodegeneration and neuroprotective effect of vit-C were measured by using 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide assay, Western blot analysis, which further conformed by the measurement of mitochondrial membrane potential using JC-1 detection kit and immunofluorescence analysis. The results showed that PTZ and ethanol produced extensive Bax-dependent caspase-9 and caspase-3 activation and caused neuronal apoptosis. Furthermore, the cotreatment of vit-C along with ethanol and PTZ showed significantly decreased expression of Bax, caspase-9, caspase-3, cytochrome-c, and significantly increased expression of antiapoptotic Bcl-2 protein when compared with control group. Our findings indicate that PTZ and ethanol activate an intrinsic apoptotic death program in neurons that is likely to contribute to the neuropathologic effects in fetal alcohol exposure, and vit-C can prevent some of the deleterious effects of PTZ and ethanol on the developing brain. The available experimental evidence and the safety of vit-C in pregnancy suggest the experimental use of ascorbic acid as a new and effective protective agent ethanol and PTZ-induced apoptotic neurodegeneration.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Apoptosis / drug effects*
  • Ascorbic Acid / pharmacology*
  • Blotting, Western
  • Cells, Cultured
  • Central Nervous System Depressants / toxicity
  • Ethanol / toxicity
  • Female
  • Fetal Alcohol Spectrum Disorders / prevention & control
  • Fluorescent Antibody Technique
  • GABA Antagonists / toxicity
  • Hippocampus / drug effects
  • Hippocampus / pathology
  • Nerve Degeneration / prevention & control*
  • Neurons / drug effects*
  • Pentylenetetrazole / toxicity
  • Pregnancy
  • Rats
  • Rats, Sprague-Dawley
  • Vitamins / pharmacology*


  • Central Nervous System Depressants
  • GABA Antagonists
  • Vitamins
  • Ethanol
  • Ascorbic Acid
  • Pentylenetetrazole