Lab Anim Res.  2010 Mar;26(1):69-74. 10.5625/lar.2010.26.1.69.

Proteomic Analysis of Hepatic Ischemia and Reperfusion Injury in Mice

• Cho EH
• Sung JH
• Koh PO

• KMID: 1458942
• DOI: http://doi.org/10.5625/lar.2010.26.1.69

Abstract

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.

Keyword

Ischemia; liver; reperfusion

MeSH Terms

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
Annexin A4
Estrogens, Conjugated (USP)
Eukaryotic Initiation Factor-3
Lactoylglutathione Lyase
Peptide Initiation Factors
Proteasome Endopeptidase Complex
Proteins
Salicylamides
Spermidine Synthase

Figure

• 

  Figure 1. Serum ALT and AST levels in sham-operated (sham) and ischemia/reperfusion-operated (I/R) animals. ALT and AST levels were significantly increased in I/R-operated animals. Data (n=5) are represented as mean±SEM. ∗P<0.01.

• 

  Figure 2. Histopathological photos of the liver in sham-operated (A) and ischemia/reperfusion-operated (B) animals. There is no lesion in the normal liver of sham-operated animals. Ischemic hepatic lobes of I/R-operated animals showed the hepatic lesion with nuclear condensation (arrows) and sinusoidal congestion (open arrows). Hematoxylin and Eosin stain. Scale bar=100 µm.

• 

  Figure 3. Two-dimensional SDS-PAGE analysis of proteins in liver tissue of sham-operated (A) and ischemia/reperfusion-operated (B) animals. Isoelectric focusing was performed at pH 4-7 using IPG strips, followed by second-dimensional separation on 7.5-17.5% gradient SDS gels stained with silver. Squares indicate the protein spots that were differentially expressed between sham-and I/R-operated animals.

• 

  Figure 4. Western blot analysis of dynamin-1-like protein (DLP-1) in liver tissue of sham-operated and ischemia/reperfusion-operated animals. Each lane represents an individual experimental animal. Densitometric analysis is represented as an arbitrary unit (A.U.), normalized by tubulin. Data (n=5) are represented as mean±SEM. ∗P<0.05.

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