Protective effects of cyclosporine and its analog NIM-811 in a murine model of hepatic ischemia-reperfusion injury^☆

Abstract

Background and aim: The liver is susceptible to ischemia-reperfusion injury (IRI) during hepatic surgery, when the vessels are compressed to control bleeding, or liver transplantation, when there is an obligate period of ischemia. The hallmark of IRI comprises mitochondrial dysfunction, which generates reactive oxygen species, and cell death through necrosis or apoptosis. Cyclosporine (CsA), which is a well-known immunosuppressive agent that inhibits calcineurin, has the additional effect of inhibiting the mitochondrial permeability transition pore (mPTP), thereby, preventing mitochondrial swelling and injury. NIM-811, which is the nonimmunosuppressive analog of CsA, has a similar effect on the mPTP. In this study, we tested the effect of both agents on mitigating warm hepatic IRI in a murine model.

Materials and methods: Before ischemic insult, the mice were administered with intraperitoneal normal saline (control); CsA at 2.5, 10, or 25 mg/kg; or NIM-811 at 10 mg/kg. Thereafter, the mice were subjected to partial warm hepatic ischemia by selective pedicle clamping for 60 min, followed by 6 h of recovery after reperfusion. Serum alanine transaminase (ALT) was measured, and the liver tissue was examined histologically for the presence of apoptosis and the levels of inflammatory cytokines.

Results: Compared with the control mice, the mice treated with 10 and 25 mg/kg of CsA and NIM-811 had significantly lower ALT levels (P < 0.001, 0.007, and 0.031, respectively). Moreover, the liver tissue showed reduced histological injury scores after treatment with CsA at 2.5, 10, and 25 mg/kg and NIM-811 (P = 0.041, <0.001, 0.003, and 0.043, respectively) and significant decrease in apoptosis after treatment with CsA at all doses (P = 0.012, 0.007, and <0.001, respectively). Levels of the pro-inflammatory cytokines, particularly interleukin (IL)-1β, IL-2, IL-4, IL-10, and keratinocyte chemoattractant/human growth-regulated oncogene significantly decreased in the mice treated with the highest dose of CsA (25 mg/kg) than those in the control mice.

Conclusions: Premedication with CsA or NIM-811 mitigated hepatic IRI in mice, as evidenced by the decreased ALT and reduced injury on histology. These results have potential implications on mitigating IRI during liver transplantation and resection.

Keywords

Cyclosporine (CsA) / Cyclosporine analogue / Liver surgery / Ischemia-reperfusion injury (IRI) / Animal model / NIM-811

Data availability statement

The data presented in this study are available from the authors upon request to the corresponding author. The study protocols and standard operating procedures are also available upon request. The data must not be processed for purposes other than statistical and scientific studies.

Authors’ contributions

Joshua Hefler contributed to research design, performance of the research, data analysis, and writing of the manuscript. Rena Pawlick, Braulio A. Marfil-Garza, Nerea Cuesta-Gomez, and Sanaz Hatami contributed to performance of the research, data analysis, and editing of the manuscript. Aducio Thiesen contributed to data analysis and editing of the manuscript. Darren H. Freed, Con-stantine Karvellas, and David L. Bigam contributed to research design and editing of the manuscript. A.M. James Shapiro contributed to research design, supervision, and editing of the manuscript. All authors read and approved the final version of this manuscript.

Declaration of competing interest

A.M. James Shapiro reports a consulting or advisory role with ViaCyte Inc., Hemostemix Inc., and Aspect Biosystems Ltd. All remaining authors declare no conflicts of interest. The authors have no other relevant affiliations or financial involvement with any organization or entity with a financial interest in or financial con-flict with the subject matter or materials discussed in the manu-script apart from those disclosed. No writing assistance was utilized in the production of this manuscript.

Acknowledgements

This research did not receive any specific grant from funding agencies in the public, commercial, or not-for-profit sectors.

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