Formation of 4-hydroxynonenal adducts with cytochrome c oxidase in rats following short-term ethanol intake.
This study addresses the role of the lipid peroxidation product, 4-hydroxynonenal (HNE), in ethanol-related damage of cytochrome c oxidase (COX) in vivo. It utilizes an animal model with acute ethanol exposure in which HNE levels in liver mitochondria are strikingly increased. Pregnant female Sprague-Dawley rats were administered 5 doses of ethanol (4 gm/kg, po at 12-hour intervals) beginning on day 17 of gestation and were sacrificed on day 19. Controls were pair-fed and received dextrose isocaloric to ethanol. Mitochondria were isolated from maternal and fetal livers and COX activities were measured spectrophotometrically. Compared with the pair-fed controls, COX activity was decreased with exposure to ethanol by 25% in maternal rats and 43% in fetal rats (P<.05). Western Blot with an HNE-Histidine antibody showed enhanced formation of HNE adducts with COX from ethanol-exposed rats, which was more pronounced in fetal than in adult livers. The HNE adducts were mainly with subunit IV of COX. The cause and effect relationship between HNE adduct formation and COX inhibition was examined in vitro by incubating purified COX with HNE. COX inhibition was accompanied by concentration-dependent HNE adduct formation that was consistent with those found in in vivo ethanol-exposed samples. These results suggest that the ethanol-related decreases in COX activity found in liver mitochondria could be attributable to HNE adduct formation with the enzyme complex. This could be an important mechanism by which modification of proteins occur in in vivo oxidative stress.
Chen
J
J
the Division of Gastroenterology and Nutrition of the Department of Medicine, the University of Texas Health Science Center at San Antonio, San Antonio, Texas, USA. chenj1@uthcsa.edu
Robinson
N C
NC
Schenker
S
S
Frosto
T A
TA
Henderson
G I
GI
D000435:Alcoholic Intoxication / Q000378:metabolism*; D000447:Aldehydes / Q000378:metabolism*; D000818:Animals; D001665:Binding Sites; D003576:Electron Transport Complex IV / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D005260:Female; D005333:Fetus / Q000378:metabolism*; D006639:Histidine / Q000378:metabolism; D007700:Kinetics; D008099:Liver / Q000196:embryology; D046911:Macromolecular Substances; D008930:Mitochondria, Liver / Q000201:enzymology*; D011247:Pregnancy; D011248:Pregnancy Complications / Q000378:metabolism*; D051381:Rats; D017207:Rats, Sprague-Dawley
D000447:Aldehydes; D046911:Macromolecular Substances; D006639:Histidine; D003576:Electron Transport Complex IV; C027576:4-hydroxy-2-nonenal