Radiochemical quantitation of conjugated dienes in rat hepatocytes exposed to oxyradicals.
Conjugated dienes are fingerprint signatures of oxidant damage in cells. We used a radiochemical method based on the Diels-Alder reaction of 14C-labeled tetracyanoethylene with conjugated dienes to delineate the changes of its levels in ischemia-reperfusion in the rat liver. To more directly illustrate the kinetics of diene appearance in hepatocytes, we have applied the same radiochemical assay to rat hepatocytes exposed to xanthine oxidase and hypoxanthine. We observed that the conjugated dienes rose to a maximum under our condition at approximately 10 min, while Trolox--an antioxidant derived from vitamin E found previously to protect rat hepatocytes from oxyradical damage (2)--markedly reduced the formation of conjugated dienes.
Wu
J
J
Department of Clinical Biochemistry, University of Toronto, Ontario, Canada.
Sugiyama
H
H
Fung
K P
KP
Carey
D
D
Wu
T W
TW
D000466:Alkadienes / Q000032:analysis*; D000818:Animals; D000975:Antioxidants / Q000494:pharmacology; D005030:Ethylenes / Q000032:analysis; D005609:Free Radicals / Q000494:pharmacology; D019271:Hypoxanthine; D007042:Hypoxanthines / Q000494:pharmacology; D008099:Liver / Q000737:chemistry* / Q000166:cytology; D008297:Male; D009570:Nitriles / Q000032:analysis; D010100:Oxygen / Q000494:pharmacology*; D011854:Radiochemistry / Q000379:methods; D051381:Rats; D017207:Rats, Sprague-Dawley; D013997:Time Factors; D014969:Xanthine Oxidase / Q000494:pharmacology
D000466:Alkadienes; D000975:Antioxidants; D005030:Ethylenes; D005609:Free Radicals; D007042:Hypoxanthines; D009570:Nitriles; D019271:Hypoxanthine; C003393:tetracyanoethylene; D014969:Xanthine Oxidase; D010100:Oxygen