Measurement of intracellular biomolecular oxidation in liver ischemia-reperfusion injury via immuno-spin trapping.
Hepatic ischemia-reperfusion (I/R) can lead to liver failure in association with remote organ damage, both of which have significant rates of morbidity and mortality. In this study, novel spin trapping and histopathological techniques have been used to investigate in vivo free radical formation in a rat model of warm liver I/R injury. 5,5-Dimethyl-1-pyrroline N-oxide (DMPO) was administered to rats via intraperitoneal injection at a single dose of 1.5g of pure DMPO/kg body wt 2h before the initiation of liver ischemia. Blood vessels supplying the median and left lateral hepatic lobes were occluded with an arterial clamp for 60min, followed by 60min reperfusion. The effects of DMPO on I/R injury were evaluated by assessing the hepatic ultrastructure via transmission electron microscopy and by histopathological scoring. Immunoelectron microscopy was performed to determine the cellular localization of DMPO nitrone adducts. Levels of nitrone adducts were also measured to determine in situ scavenging of protein and DNA radicals. Total histopathological scoring of cellular damage was significantly decreased in hepatic I/R injury after DMPO treatment. DMPO treatment significantly decreased the hepatic conversion of xanthine oxidase and 4-hydroxynonenal formation in I/R injury compared to the untreated I/R group. The distribution of gold-nanoparticle-labeled DMPO nitrone adducts was observed in mitochondria, cytoplasm, and nucleus of hepatocytes. The formation of protein- and DNA-nitrone adducts was increased in DMPO-treated I/R livers compared to DMPO controls, indicating increased in situ protein and DNA radical formation and scavenging by DMPO. These results suggest that DMPO reduces I/R damage via protection against oxidative injury.
Department of Medical Biochemistry, Akdeniz University Medical School, Antalya, Turkey.
Serdar
S
Dogan
Gulsum
G
Ozlem Elpek
Esma
E
Kirimlioglu Konuk
Nejdet
N
Demir
Mutay
M
Aslan
D000410:Alanine Transaminase / Q000097:blood; D000818:Animals; D003497:Cyclic N-Oxides / Q000008:administration & dosage* / Q000493:pharmacokinetics; D016166:Free Radical Scavengers / Q000008:administration & dosage* / Q000493:pharmacokinetics; D005609:Free Radicals; D007150:Immunohistochemistry; D007511:Ischemia / Q000201:enzymology / Q000378:metabolism / Q000473:pathology; D015227:Lipid Peroxidation; D008099:Liver / Q000098:blood supply / Q000378:metabolism* / Q000473:pathology; D008297:Male; D008315:Malondialdehyde / Q000378:metabolism; D046529:Microscopy, Electron, Transmission; D010084:Oxidation-Reduction; D051381:Rats; D017208:Rats, Wistar; D015427:Reperfusion Injury / Q000097:blood / Q000188:drug therapy / Q000378:metabolism*; D019097:Spin Trapping; D014018:Tissue Distribution; D014968:Xanthine Dehydrogenase / Q000378:metabolism; D014969:Xanthine Oxidase / Q000378:metabolism
D003497:Cyclic N-Oxides; D016166:Free Radical Scavengers; D005609:Free Radicals; D008315:Malondialdehyde; C017245:5,5-dimethyl-1-pyrroline-1-oxide; D014968:Xanthine Dehydrogenase; D014969:Xanthine Oxidase; D000410:Alanine Transaminase