Basic Study
Copyright ©The Author(s) 2017. Published by Baishideng Publishing Group Inc. All rights reserved.
World J Hepatol. Dec 8, 2017; 9(34): 1261-1269
Published online Dec 8, 2017. doi: 10.4254/wjh.v9.i34.1261
Morphological alterations and redox changes associated with hepatic warm ischemia-reperfusion injury
Rim Jawad, Melroy D’souza, Lisa Arodin Selenius, Marita Wallenberg Lundgren, Olof Danielsson, Greg Nowak, Mikael Björnstedt, Bengt Isaksson
Rim Jawad, Lisa Arodin Selenius, Marita Wallenberg Lundgren, Olof Danielsson, Mikael Björnstedt, Division of Pathology, Department of Laboratory Medicine, Karolinska Institutet, Stockholm S-141 86, Sweden
Melroy D’souza, Bengt Isaksson, Department of Clinical Science, Intervention, and Technology (CLINTEC), Division of Surgery, Karolinska Institutet, Karolinska University Hospital, Huddinge, Stockholm S-141 86, Sweden
Greg Nowak, Department of Clinical Science, Intervention, and Technology (CLINTEC), Division of Transplantation Surgery, Karolinska Institutet, Karolinska University Hospital, Huddinge, Stockholm S-141 86, Sweden
Author contributions: D’souza M, Nowak G, Björnstedt M and Isaksson B contributed to concept and design; Jawad R, D’souza M and Selenius LA contributed to acquisition of data; Jawad R, D’souza M, Lundgren MW, Danielsson O, Nowak G, Björnstedt M and Isaksson B contributed to analysis and interpretation of data; Jawad R, D’souza M and Selenius LA contributed to drafting of the manuscript; Jawad R, D’souza M, Selenius LA, Lundgren MW, Danielsson O, Nowak G, Björnstedt M and Isaksson B contributed to critical revision of the manuscript for important intellectual content; Jawad R, D’souza M and Selenius LA contributed to statistical analysis.
Supported by Swedish Cancer society (Cancerfonden) and the Swedish Cancer and Allergy fund (Cancer-och Allergifonden).
Institutional review board statement: This study was reviewed and approved by the Regional Ethics Committee for human studies, Stockholm.
Informed consent statement: All biopsy specimens and patient data were taken with informed consent for participation in the study.
Conflict-of-interest statement: No conflict of interest exists.
Data sharing statement: No additional data available.
Open-Access: This article is an open-access article which was selected by an in-house editor and fully peer-reviewed by external reviewers. It is distributed in accordance with the Creative Commons Attribution Non Commercial (CC BY-NC 4.0) license, which permits others to distribute, remix, adapt, build upon this work non-commercially, and license their derivative works on different terms, provided the original work is properly cited and the use is non-commercial. See: http://creativecommons.org/licenses/by-nc/4.0/
Correspondence to: Melroy D’souza, MS, DNB MRCSEd, Department of Clinical Science, Intervention, and Technology (CLINTEC), Division of Surgery, Karolinska Institutet, Karolinska University Hospital, Huddinge, Stockholm S-141-86, Sweden. melroy.dsouza@karolinska.se
Telephone: +46-76-2884290 Fax: +46-85-8582340
Received: May 30, 2017
Peer-review started: June 1, 2017
First decision: July 20, 2017
Revised: August 10, 2017
Accepted: October 15, 2017
Article in press: October 15, 2017
Published online: December 8, 2017
Processing time: 188 Days and 9.8 Hours
Abstract
AIM

To study the effects of warm ischemia-reperfusion (I/R) injury on hepatic morphology at the ultrastructural level and to analyze the expression of the thioredoxin (TRX) and glutaredoxin (GRX) systems.

METHODS

Eleven patients undergoing liver resection were subjected to portal triad clamping (PTC). Liver biopsies were collected at three time points; first prior to PTC (baseline), 20 min after PTC (post-ischemia) and 20 min after reperfusion (post-reperfusion). Electron microscopy and morphometry were used to study and quantify ultrastructural changes, respectively. Additionally, gene expression analysis of TRX and GRX isoforms was performed by quantitative PCR. For further validation of redox protein status, immunogold staining was performed for the isoforms GRX1 and TRX1.

RESULTS

Post-ischemia, a significant loss of the liver sinusoidal endothelial cell (LSEC) lining was observed (P = 0.0003) accompanied by a decrease of hepatocyte microvilli in the space of Disse. Hepatocellular morphology was well preserved apart from the appearance of crystalline mitochondrial inclusions in 7 out of 11 patients. Post-reperfusion biopsies had similar features as post-ischemia with the exception of signs of a reactivation of the LSECs. No changes in the expression of redox-regulatory genes could be observed at mRNA level of the isoforms of the TRX family but immunoelectron microscopy indicated a redistribution of TRX1 within the cell.

CONCLUSION

At the ultrastructural level, the major impact of hepatic warm I/R injury after PTC was borne by the LSECs with detachment and reactivation at ischemia and reperfusion, respectively. Hepatocytes morphology were well preserved. Crystalline inclusions in mitochondria were observed in the hepatocyte after ischemia.

Keywords: Hepatic ischemia-reperfusion injury; Ischemia reperfusion injury; Warm ischemia-reperfusion injury; Glutaredoxins; Thioredoxins; Electron microscopy; Oxidative stress; Portal triad clamping

Core tip: The complex mechanisms of warm Ischemia reperfusion (I/R) injury in the liver are diverse and have been widely studied but poorly understood. This study aims to investigate the ultrastructural changes at warm I/R injury induced by portal triad clamping. The effects were mainly borne by the liver sinusoidal endothelial cells (LSEC) which detached from the sinusoidal wall after ischemia. Interestingly we found that the LSECs reattached after reperfusion. Hepatocytes were unaffected except for the appearance of crystalline inclusions in the mitochondria. Investigation of redox related proteins showed no changes within our time frame.