Meguro Makoto, Katsuramaki Tadashi, Kimura Hitoshi, Isobe Masato, Nagayama Minoru, Kukita Kazuma, Nui Akihiro, Hirata Koichi
Department of Surgery (Section 1), Sapporo Medical University School of Medicine, Japan.
Transplantation. 2003 Mar 15;75(5):703-10. doi: 10.1097/01.TP.0000053400.42842.5C.
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.
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.
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.
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.
在肝脏缺血再灌注（I/R）损伤期间，细胞死亡的主要模式是凋亡还是坏死仍存在争议。此外，此前尚未研究过这些细胞死亡与新型诱导型一氧化氮合酶抑制剂（ONO-1714）效应之间的相关性。
猪在体外循环下接受180分钟的肝脏温热I/R。对照组未给予ONO-1714。在ONO-1714组中，在缺血前5分钟通过门静脉导管以0.05mg/kg的剂量给予ONO-1714。再灌注后通过末端脱氧核苷酸转移酶介导的dUTP缺口末端标记和苏木精-伊红染色检查凋亡和坏死变化。通过免疫组织化学检查硝基酪氨酸、活性半胱天冬酶-3和细胞色素c。还检测了血浆NO + NO、天冬氨酸转氨酶和乳酸脱氢酶水平。
在对照组中，凋亡细胞的频率仅为2.6%；然而，再灌注后24小时坏死细胞的频率为37%。ONO-1714显著减轻了再灌注期的凋亡和坏死、硝基酪氨酸的表达以及血浆天冬氨酸转氨酶、乳酸脱氢酶和NO(2)- + NO(3)-水平的升高。
肝脏温热I/R损伤期间细胞死亡的主要模式是坏死，凋亡不占主导。这些坏死变化是由过氧亚硝酸盐的过量产生引起的，并且ONO-1714由于抑制了过氧亚硝酸盐的产生而大大减轻了I/R损伤。
