Apoptosis and necrosis after warm ischemia-reperfusion injury of the pig liver and their inhibition by ONO-1714.
BACKGROUND
It is still controversial whether a major mode of cell death during hepatic ischemia-reperfusion (I/R) injuries is apoptosis or necrosis. Moreover, the correlation between these cell deaths and the effects of a novel inducible nitric oxide synthase inhibitor (ONO-1714) has not been studied before.


METHODS
Pigs were subjected to 180 min of hepatic warm I/R under extracorporeal circulation. The control group was not administered ONO-1714. In the ONO-1714 group, ONO-1714 was administered 5 min before ischemia at a dose of 0.05 mg/kg through a portal vein catheter. The apoptotic and necrotic changes after reperfusion were examined by terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling and hematoxylin-eosin staining. Nitrotyrosine, active caspase-3, and cytochrome c were examined by immunohistochemistry. The plasma NO + NO, aspartate aminotransferase, and lactate dehydrogenase levels were also examined.


RESULTS
In the control group, the frequency of apoptotic cells was only 2.6%; nevertheless, that of necrotic cells was 37% at 24 hr after reperfusion. ONO-1714 significantly attenuated apoptosis and necrosis, the expression of nitrotyrosine, and the increases of the plasma aspartate aminotransferase, lactate dehydrogenase, and NO(2)- + NO(3)- levels in the reperfusion phase.


CONCLUSIONS
A major mode of cell death during hepatic warm I/R injury was necrosis, and apoptosis was not dominant. These necrotic changes were caused by the excess production of peroxynitrite, and ONO-1714 greatly attenuated I/R injuries as the result of inhibition of the peroxynitrite production.
Meguro
Makoto
M
Department of Surgery (Section 1), Sapporo Medical University School of Medicine, Japan. meguro@sapmed.ac.jp
Katsuramaki
Tadashi
T
Kimura
Hitoshi
H
Isobe
Masato
M
Nagayama
Minoru
M
Kukita
Kazuma
K
Nui
Akihiro
A
Hirata
Koichi
K
D000578:Amidines / Q000494:pharmacology; D000818:Animals; D017209:Apoptosis; D001219:Aspartate Aminotransferases / Q000097:blood; D004247:DNA / Q000235:genetics; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005260:Female; D022781:Hepatocytes / Q000473:pathology; D006574:Heterocyclic Compounds, 2-Ring / Q000494:pharmacology; D006358:Hot Temperature; D020287:In Situ Nick-End Labeling; D007511:Ischemia / Q000473:pathology* / Q000503:physiopathology*; D007770:L-Lactate Dehydrogenase / Q000097:blood; D008099:Liver / Q000378:metabolism; D008102:Liver Circulation; D018613:Microscopy, Confocal; D008854:Microscopy, Electron; D009336:Necrosis; D009566:Nitrates / Q000097:blood; D019001:Nitric Oxide Synthase / Q000037:antagonists & inhibitors; D009573:Nitrites / Q000097:blood; D015427:Reperfusion Injury / Q000473:pathology* / Q000503:physiopathology*; D013552:Swine; D013997:Time Factors; D014443:Tyrosine / Q000031:analogs & derivatives* / Q000378:metabolism
C407360:7-chloro-3-imino-5-methyl-2-azabicyclo(4.1.0)heptane; D000578:Amidines; D004791:Enzyme Inhibitors; D006574:Heterocyclic Compounds, 2-Ring; D009566:Nitrates; D009573:Nitrites; C002744:3-nitrotyrosine; D014443:Tyrosine; D004247:DNA; D007770:L-Lactate Dehydrogenase; D019001:Nitric Oxide Synthase; D001219:Aspartate Aminotransferases