Role of free radicals in primary nonfunction of marginal fatty grafts from rats treated acutely with ethanol.
Acute treatment with one large dose of ethanol, which mimics binge drinking, causes marginal fatty liver and decreases survival significantly after liver transplantation in rats, yet mechanisms remain unclear. Therefore, we evaluated the possible role of free radicals in primary nonfunction caused by acute ethanol. Female donor rats were administered ethanol (5 g/kg orally) 20 hr before explantation, and grafts were stored in UW cold storage solution for 24-42 hr before implantation. Free radicals were trapped with alpha-(4-pyridyl 1-oxide)-N-tert-butylnitrone after transplantation, and adducts were detected using electron spin resonance spectrometry. Ethanol increased a carbon-centered radical adduct in bile approximately 2-fold and elevated serum lipid hydroperoxides approximately 4-fold. Ethanol also increased transaminase release 3.7-fold and decreased bile production by 55%. Catechin, a free radical scavenger, minimized the increase in free radicals, blunted transaminase release, and elevated bile production significantly, indicating that free radical production plays an important role in ethanol-induced fatty graft injury. GdCl3 (20 mg/kg intravenously), a selective Kupffer cell toxicant, largely blocked the increases in free radical and lipid hydroperoxide production caused by ethanol. In addition, ethanol nearly doubled white blood cell adhesion after transplantation, leading to increased superoxide production in fatty grafts. GdCl3 largely blocked leukocyte adhesion as well as superoxide production. Allopurinol, an inhibitor of xanthine oxidase, also diminished free radical production, blunted transaminase release, and improved bile production in fatty grafts significantly. Taken together, we conclude that free radical formation increases in ethanol-induced fatty grafts due mainly to activation of Kupffer cells and increased adhesion of white blood cells. Antioxidants can effectively block free radical formation and minimize injury to marginal fatty grafts caused by binge drinking.
Zhong
Z
Z
Department of Pharmacology, University of North Carolina, Chapel Hill, North Carolina 27599-7365, USA.
Connor
H
H
Stachlewitz
R F
RF
Frankenberg
M
M
Mason
R P
RP
Lemasters
J J
JJ
Thurman
R G
RG
D000818:Animals; D000975:Antioxidants / Q000494:pharmacology; D001646:Bile / Q000378:metabolism; D002392:Catechin / Q000494:pharmacology; D002448:Cell Adhesion / Q000187:drug effects; D000431:Ethanol / Q000494:pharmacology*; D005260:Female; D005609:Free Radicals / Q000378:metabolism; D006085:Graft Survival / Q000187:drug effects*; D006861:Hydrogen Peroxide / Q000378:metabolism; D007728:Kupffer Cells / Q000187:drug effects* / Q000201:enzymology; D007962:Leukocytes / Q000187:drug effects / Q000502:physiology; D015227:Lipid Peroxidation; D016031:Liver Transplantation; D008315:Malondialdehyde / Q000378:metabolism; D008930:Mitochondria, Liver / Q000187:drug effects / Q000378:metabolism; D051381:Rats; D013481:Superoxides / Q000378:metabolism; D014969:Xanthine Oxidase / Q000037:antagonists & inhibitors
D000975:Antioxidants; D005609:Free Radicals; D013481:Superoxides; D000431:Ethanol; D008315:Malondialdehyde; D002392:Catechin; D006861:Hydrogen Peroxide; D014969:Xanthine Oxidase