J Korean Surg Soc.  2012 Nov;83(5):298-306. 10.4174/jkss.2012.83.5.298.

Immediately transcripted genes in various hepatic ischemia models

• Choi KK
• Cho JA
• Kim SH
• Lee SW
• Min SO
• Kim KS

• KMID: 2145007
• DOI: http://doi.org/10.4174/jkss.2012.83.5.298

Abstract

PURPOSE
To elucidate the characteristic gene transcription profiles among various hepatic ischemia conditions, immediately transcribed genes and the degree of ischemic injury were compared among total ischemia (TI), intermittent clamping (IC), and ischemic preconditioning (IPC).
METHODS
Sprague-Dawley rats were equally divided into control (C, sham-operated), TI (ischemia for 90 minutes), IC (ischemia for 15 minutes and reperfusion for 5 minutes, repeated six times), and IPC (ischemia for 15 minutes, reperfusion for 5 minutes, and ischemia again for 90 minutes) groups. A cDNA microarray analysis was performed using hepatic tissues obtained by partial hepatectomy after occluding hepatic inflow.
RESULTS
The cDNA microarray revealed the following: interleukin (IL)-1beta expression was 2-fold greater in the TI group than in the C group. In the IC group, IL-1alpha/beta expression increased by 2.5-fold, and Na+/K+ ATPase beta1 expression decreased by 2.4-fold. In the IPC group, interferon regulatory factor-1, osteoprotegerin, and retinoblastoma-1 expression increased by approximately 2-fold compared to that in the C group, but the expression of Na+/K+ ATPase beta1 decreased 3-fold.
CONCLUSION
The current findings revealed characteristic gene expression profiles under various ischemic conditions. However, additional studies are needed to clarify the mechanism of protection against IPC.

Keyword

Reperfusion injury; Ischemic preconditioning; Necrosis; Apoptosis; Microarray analysis

MeSH Terms

Adenosine Triphosphatases
Apoptosis
Constriction
Hepatectomy
Interferon Regulatory Factor-1
Interleukins
Ischemia
Ischemic Preconditioning
Microarray Analysis
Necrosis
Oligonucleotide Array Sequence Analysis
Osteoprotegerin
Rats, Sprague-Dawley
Reperfusion
Reperfusion Injury
Transcriptome
Adenosine Triphosphatases
Interferon Regulatory Factor-1
Interleukins
Osteoprotegerin

Figure

• 

  Fig. 1 Experimental design. The white area represents reperfusion time, and the black area represents ischemia time. C, control; TI, total ischemia; IC, intermittent clamping; IPC, ischemic preconditioning. The C group underwent sham operations (laparotomy with exposure of the liver but without vascular occlusion), the TI group underwent continuous clamping with 90 minutes of ischemia, the IC group underwent six cycles of intermittent clamping with 15 minutes of ischemia and 5 minutes of reperfusion, and the IPC group underwent clamping with 90 minutes of ischemia after IPC (15 minutes of clamping followed by 5 minutes reperfusion).

• 

  Fig. 2 Hematoxylin-eosin staining of resected livers (×40 or ×400 magnification). The degree of liver damage was determined at ×400 magnification by using a point-counting method as follows: grade 0, minimal or no evidence of injury; grade 1, mild injury consisting of cytoplasm vacuolation and focal nuclear pyknosis; grade 2, moderate to severe injury with extensive nuclear pyknosis, cytoplasmichypereosinophilia, and loss of intercellular borders; grade 3, severe necrosis with disintegration of hepatic cords, hemorrhage, and neutrophil infiltration. The C group had minimal or no evidence of injury (grade 0). All three treatment groups exhibited grade 1 liver damage. C, control; TI, total ischemia; IC, intermittent clamping; IPC, ischemic preconditioning.

• 

  Fig. 3 Terminal deoxyuridine triphosphate nick end labeling (TUNEL) staining (×40 or ×400 magnification). The TI group had significantly more TUNEL-positive hepatocytes than the C and IC groups. There were no significant differences in TUNEL-positive hepatocytes among the C, IC, and IPC groups. C, control; TI, total ischemia; IC, intermittent clamping; IPC, ischemic preconditioning.

• 

  Fig. 4 Number of terminal deoxyuridine triphosphate nick end labeling (TUNEL)-positive cells. The number of TUNEL-positive cells was significantly higher in the experimental groups than in the C group. Among the experimental groups, the total ischemia (TI) group had significantly more positive hepatocytes than the other two groups. IC, intermittent clamping; IPC, ischemic preconditioning. *Statistical significance was tested using Kruskal-Wallis test of variance among groups. The comparison between the groups was performed using Tukey's post hoc test.

• 

  Fig. 5 Hierarchical clustering of the intermittent clamping (IC), ischemic preconditioning (IPC), and total ischemia (TI) groups. The hierarchical clustering of the IC, IPC, and TI group reveals different genetic expression patterns among the groups regarding immediate gene transcription in the livers of rats.

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