Formation of 8-hydroxy-2'-deoxyguanosine and 4-hydroxy-2-nonenal-modified proteins in rat liver after ischemia-reperfusion: distinct localization of the two oxidatively modified products.
Ischemia-reperfusion (IR) injury is an intractable process associated not only with therapeutic recanalization of vessels, but also with partial resection or transplantation of solid organs including liver. To develop methods for predicting the degree of hepatic IR injury and further to identify injured cells, we studied the formation of 8-hydroxy-2'-deoxy-guanosine (8-OHdG) and 4-hydroxy-2-nonenal (HNE)-modified proteins in the normothermic hepatic IR model of rats using immunohistochemistry, high-performance liquid chromatography (HPLC) determination and Western blot. The Pringle maneuver for either 15 or 30 min duration produced reversible or lethal damage, respectively. The levels of both products were significantly increased in proportion to ischemia duration 40 min after reperfusion, suggesting the involvement of hydroxyl radicals. Increased immunoreactivity of 8-OHdG was observed not only in the nuclei of hepatocytes but also in those of bile canalicular and endothelial cells. However, immunoreactivity of HNE-modified proteins was detected in the cytoplasm of hepatocytes, which was confirmed by Western blot, and in addition, in the nuclei of hepatocytes after severe injury. Thus, localization of the two oxidatively modified products was not identical. Our data suggest that these two products could be used for the assessment of hepatic IR injury in tissue, but that the biological significance of the two products might be different.
Department of Gastroenterological Surgery, Graduate School of Medicine, Kyoto University, Japan.
K
K
Yamagami
Y
Y
Yamamoto
M
M
Kume
Y
Y
Ishikawa
Y
Y
Yamaoka
H
H
Hiai
S
S
Toyokuni
D000080242:8-Hydroxy-2'-Deoxyguanosine; D000447:Aldehydes / Q000032:analysis*; D000818:Animals; D015153:Blotting, Western; D001831:Body Temperature; D002851:Chromatography, High Pressure Liquid; D018736:DNA Adducts / Q000032:analysis*; D004249:DNA Damage; D003849:Deoxyguanosine / Q000031:analogs & derivatives* / Q000032:analysis*; D022781:Hepatocytes / Q000378:metabolism*; D017665:Hydroxyl Radical; D007124:Immunoenzyme Techniques; D007511:Ischemia / Q000378:metabolism*; D008099:Liver / Q000098:blood supply / Q000378:metabolism*; D008111:Liver Function Tests; D008297:Male; D008970:Molecular Weight; D009687:Nuclear Proteins / Q000737:chemistry; D010084:Oxidation-Reduction; D018384:Oxidative Stress; D011506:Proteins / Q000737:chemistry*; D051381:Rats; D017208:Rats, Wistar; D015427:Reperfusion Injury / Q000378:metabolism*; D013047:Specific Pathogen-Free Organisms; D013997:Time Factors
D000447:Aldehydes; D018736:DNA Adducts; D009687:Nuclear Proteins; D011506:Proteins; D017665:Hydroxyl Radical; D000080242:8-Hydroxy-2'-Deoxyguanosine; D003849:Deoxyguanosine; C027576:4-hydroxy-2-nonenal