Markers of oxidative stress in placental villi exposed to ethanol.
OBJECTIVE
Ethanol exposure during pregnancy may result in fetal alcohol syndrome (FAS). The mechanism by which this occurs is unknown. Recent studies in several organ systems, including the placenta, suggest that oxidative stress is involved. In this study we investigated the presence and levels of three oxidative stress markers in placental villous tissue exposed to ethanol.


METHODS
Villous tissues from normal placentas were perfused with Dulbeco's modified Eagle's medium (DMEM) with HEPES buffer, sodium bicarbonate, and glucose at pH 7.4. After stabilization, 100 mM ethanol was added to the perfusate. After 2 hours of perfusion, the tissue was removed, fixed and stained for nitrotyrosine, 4-hydroxy-2-nonenal (4HNE) and 8-hydroxyguanosine (8-OHDG). Staining within the trophoblasts was quantified with densitometry.


RESULTS
Nitrotyrosine and 4HNE immunostaining was seen in the trophoblasts. 4HNE was also seen in the stroma. In contrast, 8-OHDG was seen only in the stroma and endothelial cells in the fetal circulation. Ethanol exposure significantly increased nitrotyrosine levels in the trophoblasts beyond levels in the control tissue. Nitrotyrosine and 8-OHDG levels were also increased in stroma.


CONCLUSIONS
Within the placental villi, markers of oxidative stress are present in the trophoblasts and stroma after a short period of ethanol exposure. There is an increase in oxidative stress, primarily involving the nitric oxide pathway, in the trophoblasts as well as DNA damage in the stroma. Lipid peroxidation is not acutely changed in our 2-hour exposure window.
Kay
Helen H
HH
Department of Obstetrics and Gynecology, University of Arkansas for Medical Sciences, Little Rock, Arkansas 722205, USA. kayhelenh@uams.edu
Tsoi
Stephen
S
Grindle
Kreg
K
Magness
Ronald R
RR
D000447:Aldehydes / Q000032:analysis; D015415:Biomarkers / Q000032:analysis*; D000431:Ethanol / Q000494:pharmacology*; D005260:Female; D006151:Guanosine / Q000031:analogs & derivatives / Q000032:analysis; D006801:Humans; D018384:Oxidative Stress; D010920:Placenta / Q000737:chemistry* / Q000187:drug effects*; D011247:Pregnancy; D014327:Trophoblasts / Q000737:chemistry; D014443:Tyrosine / Q000031:analogs & derivatives / Q000032:analysis
D000447:Aldehydes; D015415:Biomarkers; D006151:Guanosine; C002744:3-nitrotyrosine; C046215:8-hydroxyguanosine; D000431:Ethanol; D014443:Tyrosine; C027576:4-hydroxy-2-nonenal