Basic Study
Copyright ©The Author(s) 2017. Published by Baishideng Publishing Group Inc. All rights reserved.
World J Gastroenterol. Feb 7, 2017; 23(5): 830-841
Published online Feb 7, 2017. doi: 10.3748/wjg.v23.i5.830
Remote ischemic perconditioning prevents liver transplantation-induced ischemia/reperfusion injury in rats: Role of ROS/RNS and eNOS
Ning He, Jun-Jun Jia, Jian-Hui Li, Yan-Fei Zhou, Bing-Yi Lin, Yi-Fan Peng, Jun-Jie Chen, Tian-Chi Chen, Rong-Liang Tong, Li Jiang, Hai-Yang Xie, Lin Zhou, Shu-Sen Zheng
Ning He, Jun-Jun Jia, Li Jiang, Yan-Fei Zhou, Bing-Yi Lin, Yi-Fan Peng, Jun-Jie Chen, Tian-Chi Chen, Rong-Liang Tong, Jian-Hui Li, Hai-Yang Xie, Lin Zhou, Shu-Sen Zheng, Key Lab of Combined Multi-Organ Transplantation, Ministry of Public Health, Hangzhou 310003, Zhejiang Province, China
Jian-Hui Li, Hai-Yang Xie, Lin Zhou, Shu-Sen Zheng, Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases, Hangzhou 310003, Zhejiang Province, China
Shu-Sen Zheng, Division of Hepatobiliary and Pancreatic Surgery, Department of Surgery, First Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou 310003, Zhejiang Province, China
Author contributions: He N and Jia JJ contributed equally to this work; Zhou L and Zheng SS conceived the experiments; He N, Jia JJ, Li JH, Zhou YF, Lin BY, Peng YF, Chen JJ, and Jiang L conducted the experiments; Xie HY contributed reagents/materials/analysis tools; Chen TC and Tong RL analyzed the results; He N and Jia JJ wrote the paper; All authors reviewed the manuscript to be published.
Supported by National Natural Science Foundation of China, No. 81421062, the Science and Technology Bureau of Zhejiang Province, China, No. 2016C33145; the National Natural Science Foundation of China, No. 81470891, and the 863 National High Technology Research and Development Program of China for young scientist No. 2015AA020923.
Institutional review board statement: The study was reviewed and approved by the Experimental Animal Center of the First Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou, China.
Institutional animal care and use committee statement: All procedures involving animals were reviewed and approved by the Institutional Animal Care and Use Committee of the First Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou, China.
Conflict-of-interest statement: The authors declare there is no conflict of interest related to this study.
Data sharing statement: Technical appendix, statistical code and dataset are available from the corresponding author at shusenzheng@zju.edu.cn.
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: Shu-Sen Zheng, MD, PhD, FACS, Division of Hepatobiliary and Pancreatic Surgery, Department of Surgery, First Affiliated Hospital, Zhejiang University School of Medicine, No. 79 Qingchun Road, Hangzhou 310003, Zhejiang Province, China. shusenzheng@zju.edu.cn
Telephone: +86-571-87236567 Fax: +86-571-87236884
Received: September 25, 2016
Peer-review started: September 26, 2016
First decision: December 2, 2016
Revised: December 8, 2016
Accepted: December 21, 2016
Article in press: December 21, 2016
Published online: February 7, 2017
Processing time: 118 Days and 19.6 Hours
Abstract
AIM

To investigate the underlying mechanisms of the protective role of remote ischemic perconditioning (RIPerC) in rat liver transplantation.

METHODS

Sprague-Dawley rats were subjected to sham, orthotopic liver transplantation (OLT), ischemic postconditioning (IPostC) or RIPerC. After 3 h reperfusion, blood samples were taken for measurement of alanine aminotransferase, aspartate aminotransferase, creatinine (Cr) and creatinine kinase-myocardial band (CK-MB). The liver lobes were harvested for the following measurements: reactive oxygen species (ROS), H2O2, mitochondrial membrane potential (ΔΨm) and total nitric oxide (NO). These measurements were determined using an ROS/H2O2, JC1 and Total NOx Assay Kit, respectively. Endothelial NO synthase (eNOS) was analyzed by reverse transcription-polymerase chain reaction (RT-PCR) and western blotting, and peroxynitrite was semi-quantified by western blotting of 3-nitrotyrosine.

RESULTS

Compared with the OLT group, the grafts subjected to RIPerC showed significantly improved liver and remote organ functions (P < 0.05). ROS (P < 0.001) including H2O2 (P < 0.05) were largely elevated in the OLT group as compared with the sham group, and RIPerC (P < 0.05) reversed this trend. The collapse of ΔΨm induced by OLT ischemia/reperfusion (I/R) injury was significantly attenuated in the RIPerC group (P < 0.001). A marked increase of NO content and phosphoserine eNOS, both in protein and mRNA levels, was observed in liver graft of the RIPerC group as compared with the OLT group (P < 0.05). I/R-induced 3-nitrotyrosine content was significantly reduced in the RIPerC group as compared with the OLT group (P < 0.05). There were no significant differences between the RIPerC and IPostC groups for all the results except Cr. The Cr level was lower in the RIPerC group than in the IPostC group (P < 0.01).

CONCLUSION

Liver graft protection by RIPerC is similar to or better than that of IPostC, and involves inhibition of oxidative stress and up-regulation of the PI3K/Akt/eNOS/NO pathway.

Keywords: Liver transplantation; Ischemia/reperfusion injury; Remote ischemic perconditioning; Endothelial nitric oxide synthase; Reactive oxygen species

Core tip: This study is believed to be the first to investigate remote ischemic conditioning using a novel model of remote ischemic perconditioning (RIPerC) in liver transplantation and to identify the PI3K/Akt/endothelial nitric oxide (NO) synthase/NO axis involved. Compared to the traditional method of ischemic postconditioning, RIPerC works similar to or better than it and overcomes the main concern of increasing total ischemic time, which may lead to problems. RIPerC appears as the most promising technique to avoid ischemia/reperfusion injury in liver transplantation and is convenient clinically.