Hepatic ischemia/reperfusion (I/R) injury is an inevitable consequence during liver surgery. I/R injury induces serious hepatic dysfunction and failure. In this study, we identified proteins that were differentially expressed between sham and I/R injured livers. Animals were subjected to hepatic ischemia for 1 hr and were sacrificed at 3hr after reperfusion. Serum ALT and AST levels were significantly increased in I/R-operated animals compared to those of sham-operated animals. Ischemic hepatic lobes of I/R-operated animals showed the hepatic lesion with unclear condensation and sinusoidal congestion. Proteins from hepatic tissue were separated using two dimensional gel electrophosresis. Protein spots with a greater than 2.5-fold change in intensity were identified by mass spectrometry. Among these proteins, glutaredoxin-3, peroxiredoxin-3, glyoxalase I, spermidine synthase, dynamin-1-like protein, annexin A4, eukaryotic initiation factor 3, eukaryotic initiation factor 4A-I, 26S proteasome, proteasome alpha 1, and proteasome beta 4 levels were significantly decreased in I/R-operated animals compared to those of sham-operated animals. These proteins are related to protein synthesis, cellular growth and stabilization, anti-oxidant action. Moreover, Western blot analysis confirmed that dynamin-1-like protein levels were decreased in I/R-operated animals. Our results suggest that hepatic I/R induces the hepatic cells damage by regulation of several proteins.
Animals ; Annexin A4 ; Blotting, Western ; Estrogens, Conjugated (USP) ; Eukaryotic Initiation Factor-3 ; Hepatocytes ; Ischemia ; Lactoylglutathione Lyase ; Liver ; Mass Spectrometry ; Mice ; Peptide Initiation Factors ; Proteasome Endopeptidase Complex ; Proteins ; Reperfusion ; Reperfusion Injury ; Salicylamides ; Spermidine Synthase

Nitric oxide synthases (NOSs) play complex roles in the regulation of cardiac excitation contraction coupling under basal and stressed conditions. Herein, using the recording approach for intracellular calcium transient and synchronous myocyte contraction, the potential mechanism for NOSs-mediated cardiomyocyte contraction was explored. We found that selective inhibition of neuronal NOS (nNOS) with 100 µmol/L spermidine markedly enhanced the cardiomyocyte twitch [control: (10.5 ± 0.21)%; nNOS inhibition: (12.4 ± 0.18)%] and calcium transient [control: (0.27 ± 0.03)%; nNOS inhibition: (0.42 ± 0.01)%], but slowed the relengthening of twitch [control: (25.2 ± 1.3) ms; nNOS inhibition: (53 ± 2.8) ms] and the calcium transient decay [control: (129 ± 4.3) ms; nNOS inhibition: (176 ± 7.1) ms], which was similar to that by dynamin inhibition with 30 µmol/L dynasore. The nNOS inhibition- or dynasore-mediated effects could be rescued by an NO donor, S-Nitroso-N-acetylpenicillamine (SNAP). Our data suggest that the selective nNOS-mediated regulation of cardiac contractile activity may partly involve the dynamin-mediated endocytic mechanism.
Animals ; Biological Transport ; Calcium Signaling ; Dynamins ; antagonists & inhibitors ; physiology ; Endocytosis ; physiology ; Female ; Hydrazones ; pharmacology ; Male ; Myocardial Contraction ; physiology ; Nitric Oxide Synthase Type I ; physiology ; Rats ; Rats, Sprague-Dawley ; Spermidine ; pharmacology ; Transport Vesicles ; physiology