Cathepsin B inactivation attenuates the apoptotic injury induced by ischemia/reperfusion of mouse liver |
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Authors: | Email author" target="_blank">Z?Ben-AriEmail author E?Mor D?Azarov J?Sulkes R?Tor Y?Cheporko E?Hochhauser O?Pappo |
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Institution: | (1) Liver institute and Department of Medicine D, Israel;(2) Department of Transplantation, Israel;(3) Epidemiology Unit, Israel;(4) Clinical Biochemistry Laboratory, Rabin Medical Center, Tel Aviv University, Beilinson Campus, Petah Tiqva, Tel Aviv, Israel;(5) The Cardiac Research Laboratory of the Department of Cardiothoracic Surgery, Felsenstein Medical Research Center, Sackler School of Medicine, Tel Aviv University, Petah Tiqva, Tel Aviv, Israel;(6) Department of Histopathology, Rabin Medical Center, Tel Aviv University, Beilinson Campus, Petah Tiqva, Tel Aviv, Israel;(7) Liver institute and Department of Medicine D, Rabin Medical Center, Beilinson Campus, Petah Tiqva 49100, Israel |
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Abstract: | Background: A major mechanism underlying warm ischemia/reperfusion (I/R) injury during liver transplantation is the activation of the
caspase chain, which leads to apoptosis. Recently, it was demonstrated that the release of cathepsin B, a cysteine protease,
from the cytosol in liver injury induces mitochondrial release of cytochrome c and the activation of caspase-3 and -9, thereby
leading to apoptosis. The aim of this study was to ascertain if cathepsin B inactivation attenuates the apoptotic injury due
to I/R in mouse liver.
Methods: A model of segmental (70%) hepatic ischemia was used. Eighteen mice were anesthetized and randomly divided into three groups:
(1) Control group: sham operation (laparotomy); (2) Ischemic group: midline laparotomy followed by occlusion of all structures
in the portal triad to the left and median lobes for 60 min (ischemic period); (3) Study group: like group 2, but with intraperitoneal
administration of a pharmacological inhibitor of cathepsin B (4 mg/100 g) 30 min before induction of ischemia. Serum liver
enzyme levels were measured by biochemical analysis, and intrahepatic caspase-3 activity was measured by fluorometric assay;
apoptotic cells were identified by morphological criteria, the terminal deoxynucleotidyl transferase-mediated dUTP nick-end
labeling (TUNEL) fluorometric assay, and immunohistochemistry for caspase-3. Results: Showed that at 6 h of reperfusion, there was a statistically significant reduction in liver enzyme levels in the animals
pretreated with cathepsin B inhibitor (p < 0.05). On fluorometric assay, caspase-3 activity was significantly decreased in group 3 compared to group 2 (p < 0.0001). The reduction in postischemic apoptotic hepatic injury in the cathepsin B inhibitor -treated group was confirmed
morphologically, by the significantly fewer apoptotic hepatocyte cells detected (p < 0.05); immunohistochemically, by the significantly weaker activation of caspase-3 compared to the ischemic group (p < 0.05); and by the TUNEL assay (p < 0.05). Conclusion: The administration of cathepsin B inhibitor before induction of ischemia can attenuate postischemic hepatocyte apoptosis
and thereby minimize liver damage. Apoptotic hepatic injury seems to be mediated through caspase-3 activity. These findings
have important implications for the potential use of cathepsin B inhibitors in I/R injury during liver transplantation. |
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Keywords: | apoptosis cathepsin B injury ischemia/reperfusion liver rat |
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