Basic Study
Copyright ©The Author(s) 2017. Published by Baishideng Publishing Group Inc. All rights reserved.
World J Hepatol. Nov 18, 2017; 9(32): 1227-1238
Published online Nov 18, 2017. doi: 10.4254/wjh.v9.i32.1227
Liver atrophy after percutaneous transhepatic portal embolization occurs in two histological phases: Hepatocellular atrophy followed by apoptosis
Yasuhito Iwao, Hidenori Ojima, Tatsushi Kobayashi, Yoji Kishi, Satoshi Nara, Minoru Esaki, Kazuaki Shimada, Nobuyoshi Hiraoka, Minoru Tanabe, Yae Kanai
Yasuhito Iwao, Hidenori Ojima, Nobuyoshi Hiraoka, Yae Kanai, Division of Molecular Pathology, National Cancer Center Research Institute, Tokyo 104-0045, Japan
Yasuhito Iwao, Yoji Kishi, Satoshi Nara, Minoru Esaki, Kazuaki Shimada, Hepatobiliary and Pancreatic Surgery Division, National Cancer Center Hospital, Tokyo 104-0045, Japan
Yasuhito Iwao, Minoru Tanabe, Hepatobiliary and Pancreatic Surgery, Tokyo Medical and Dental University, Tokyo 113-8510, Japan
Hidenori Ojima, Yae Kanai, Department of Pathology, Keio University School of Medicine, Tokyo 160-8582, Japan
Tatsushi Kobayashi, Diagnostic Radiology Division, National Cancer Center Hospital East, Chiba 277-8577, Japan
Author contributions: Iwao Y and Ojima H performed the experiments, analyzed the data and wrote the paper; Kobayashi T obtained the pig specimens; Kishi Y, Nara S, Esaki M and Shimada K obtained the human surgical liver specimens and determined the clinical data; Hiraoka N, Tanabe M and Kanai Y coordinated the research.
Supported by National Cancer Center Research and Development Fund (23-A-35).
Institutional review board statement: All patients gave written informed consent for inclusion in this study (2007-022).
Institutional animal care and use committee statement: All animal experiment protocols were approved by the Committee for Ethics in Animal Experimentation and were conducted in accordance with the Guidelines for Animal Experiments of our institution (K03-004).
Conflict-of-interest statement: The authors have no conflict of interest to disclose.
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: Hidenori Ojima, MD, PhD, Division of Molecular Pathology, National Cancer Center Research Institute, 5-1-1, Tsukiji, Chuo-ku, Tokyo 104-0045, Japan. hojima@ncc.go.jp
Telephone: +81-3-53633764 Fax: +81-3-3353-3290
Received: August 7, 2017
Peer-review started: August 7, 2017
First decision: September 13, 2017
Revised: October 9, 2017
Accepted: October 30, 2017
Article in press: October 30, 2017
Published online: November 18, 2017
Processing time: 101 Days and 0 Hours
ARTICLE HIGHLIGHTS
Research background

The interruption of portal blood flow by portal vein embolization or tumor thrombosis, for example, causes liver atrophy. However, the mechanisms responsible for this effect have not been fully elucidated.

Research motivation

The previous study suggested that the mechanisms responsible for liver atrophy likely commence soon after the disruption of portal blood flow. Consequently, histopathological changes would likely also be observed soon after percutaneous transhepatic portal embolization (PTPE). Recently, the relationship between apoptosis and autophagy has been extensively reported. Autophagy in the liver is reportedly caused by starvation and is related to hepatocellular atrophy, and, moreover, interruption of the portal blood flow, which contains a wealth of nutrients, is considered a form of starvation. Therefore, autophagy may be related to both cellular shrinking and apoptosis. However, the relationship between portal venous obstruction and autophagy has not been reported. To clarify the mechanisms responsible for liver atrophy, histopathological analysis should be carried out repeatedly within the first few weeks after PTPE. However, to the best of our knowledge, such time-course studies have not yet been carried out. The results and hypotheses will provide a basis for understanding the mechanism of liver atrophy after interruption of the portal blood flow and will facilitate further study.

Research objectives

The aim of this study was to investigate, using specimens from a previously reported porcine PTPE model, the microscopic changes associated with apoptosis and autophagy in the days and weeks following portal venous obstruction and to clarify the mechanism by which interrupted portal blood flow causes liver atrophy. Furthermore, to understand the mechanism of liver atrophy in humans after PTPE, the authors sought to verify the integrity of the pig results by performing the same histopathological investigations in specimens resected from human patients who had undergone PTPE.

Research methods

The authors performed histopathological examinations of liver specimens from five pigs that had undergone PTPE in a time-dependent model of liver atrophy. In specimens from embolized lobes (EMB) and nonembolized lobes (controls), the authors measured the portal vein to central vein distance (PV-CV), the area and number of hepatocytes per lobule, and apoptotic activity using the terminal deoxynucleotidyl transferase dUTP nick-end labeling assay. Immunohistochemical reactivities were evaluated for light chain 3 (LC3) and lysosomal-associated membrane protein 2 (LAMP2) as autophagy markers and for glutamine synthetase and cytochrome P450 2E1 (CYP2E1) as metabolic zonation markers. Samples from ten human livers taken 20-36 d after PTPE were similarly examined.

Research results

PV-CVs and lobule areas did not differ between EMB and controls at day 0, but were lower in EMB than in controls at weeks 2, 4, and 6. Hepatocyte numbers were not significantly reduced in EMB at day 0 and week 2 but were reduced at weeks 4 and 6. Apoptotic activity was higher in EMB than in controls at day 0 and week 4. LC3 and LAMP2 staining peaked in EMB at week 2, with no significant difference between EMB and controls at weeks 4 and 6. Glutamine synthetase and CYP2E1 zonation in EMB at weeks 2, 4, and 6 were narrower than those in controls. Human results were consistent with those of porcine specimens. However the number of pigs was insufficient for a detailed histopathological study to provide unequivocal evidence of the relationship between hepatocellular atrophy and autophagy.

Research conclusions

To investigate the mechanism by which portal vein obstruction causes liver atrophy, the authors examined the histological changes in pig livers following PTPE and observed two distinct phases. The first phase, termed the hepatocellular atrophic phase, is characterized by lobular shrinkage without hepatocyte loss and with high levels of LC3 and LAMP2 expression. This phase lasted for the first 2 wk following PTPE. The second phase, which occurs between weeks 2 and 4, is termed the apoptotic phase and is characterized by a reduction in hepatocyte numbers without a reduction in lobular size. This is accompanied by reduced LC3 and LAMP2 expression and increased TUNEL staining. Human liver specimens resected after PTPE had many similar characteristics to specimens collected from pigs at week 4. Despite liver atrophy appearing to be mostly resolved 2 wk after embolization, the period after PTPE could beneficially be extended to 4 wk to ensure contralateral hypertrophy and to allow the completion of liver atrophy.

Research perspectives

Histopathological analysis is the best way to clarify the mechanisms responsible for liver atrophy. To assess microscopic changes in liver tissues, it is important to study liver lobules, the smallest functional units of the liver. The observation of clear histological changes would be expected. To clarify the more detailed mechanism of liver atrophy after interruption of the portal blood flow, the authors have to study the histopathological changes using not only the pig model but also small animal models, e.g., mouse models, because such animals are easy to handle. After such detailed studies, future research will hopefully provide a basis for understanding the mechanism of liver atrophy after interruption of the portal blood flow and also give a sound theoretical basis for planning treatment strategies for acute portal obstruction-related liver dysfunction or disease and chronic ischemic-related liver diseases with liver atrophy.