Ethanol reduces expression of apoptotic proteins after hypoxia/reoxygenation in a brain slice model.
OBJECTIVE
Acute administration of ethanol is associated with neuroprotection in rat with transient cerebral ischemia. To investigate the molecular mechanism of ethanol-induced neuroprotection, we determined the effect of ethanol on expression levels of apoptotic proteins, including  caspase-3, Bcl-2-associated X protein (Bax), and apoptosis-inducing factor (AIF). To assess overall cell viability following ethanol treatment, ADP/ATP ratio was measured.


METHODS
Brain slice cultures were prepared using postnatal 10-day-old Sprage-Dawley rats. Brain slices  were divided into control and hypoxia groups. Hypoxia groups include a non-treatment group and three treatment groups (10, 30, or 90 mM ethanol). Levels of caspase-3, Bax, and AIF were determined by western blot. ADP/ATP ratio was assessed using ADP/ATP assay kit.


RESULTS
Ethanol  administration reduced ADP/ATP ratio in all three treatment groups (10, 30, and 90 mM). A reduction in caspase-3, BAX, and AIF expression was observed with all three treatment groups in conjunction with decreased ADP/ATP levels. The three treatment groups showed similar levels of reduction  in ADP/ATP ratio and apoptotic protein expression.


CONCLUSIONS
Ethanol-induced neuroprotection involves inhibition of apoptotic pathways, including Bax, caspase-3, and AIF. Dose range of 10-90 mM ethanol provides similar level of protection compared to 10 mM ethanol.
Department of Neurological Surgery, Wayne State University School of Medicine, Detroit, MI 48201, USA.
Yu
Y
Yuan
Changya
C
Peng
Kevin
K
Li
Mohammed
M
Hussain
Chaitanya
C
Sikharam
Murali
M
Guthikonda
Yuchuan
Y
Ding
D000818:Animals; D017209:Apoptosis / Q000187:drug effects*; D051033:Apoptosis Inducing Factor / Q000378:metabolism; D015153:Blotting, Western; D001921:Brain / Q000187:drug effects; D053148:Caspase 3 / Q000378:metabolism; D002470:Cell Survival / Q000187:drug effects; D000431:Ethanol / Q000494:pharmacology*; D020925:Hypoxia-Ischemia, Brain / Q000378:metabolism*; D018696:Neuroprotective Agents / Q000494:pharmacology*; D009924:Organ Culture Techniques; D051381:Rats; D017207:Rats, Sprague-Dawley; D051028:bcl-2-Associated X Protein / Q000378:metabolism
D051033:Apoptosis Inducing Factor; D018696:Neuroprotective Agents; D051028:bcl-2-Associated X Protein; D000431:Ethanol; D053148:Caspase 3