Heavy Water (D2O) Containing Preservation Solution Reduces Hepatic Cold Preservation and Reperfusion Injury in an Isolated Perfused Rat Liver (IPRL) Model.

Shingo Shimada,Kenji Wakayama,Akinobu Taketomi,Sodai Sakamoto,Tsuyoshi Shimamura,Kengo Shibata,Masato Fujiyoshi,Takahisa Ishikawa,Norio Kawamura,Moto Fukai

doi:10.3390/jcm8111818

Shingo Shimada, Kenji Wakayama + Show 8 more

Open Access

https://doi.org/10.3390/jcm8111818

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Abstract

Background: Heavy water (D2O) has many biological effects due to the isotope effect of deuterium. We previously reported the efficacy of D2O containing solution (Dsol) in the cold preservation of rat hearts. Here, we evaluated whether Dsol reduced hepatic cold preservation and reperfusion injury. Methods: Rat livers were subjected to 48-hour cold storage in University of Wisconsin (UW) solution or Dsol, and subsequently reperfused on an isolated perfused rat liver. Graft function, injury, perfusion kinetics, oxidative stress, and cytoskeletal integrity were assessed. Results: In the UW group, severe ischemia and reperfusion injury (IRI) was shown by histopathology, higher liver enzymes leakage, portal resistance, and apoptotic index, oxygen consumption, less bile production, energy charge, and reduced glutathione (GSH)/oxidized glutathione (GSSG) ratio (versus control). The Dsol group showed that these injuries were significantly ameliorated (versus the UW group). Furthermore, cytoskeletal derangement was progressed in the UW group, as shown by less degradation of α-Fodrin and by the inactivation of the actin depolymerization pathway, whereas these changes were significantly suppressed in the Dsol group. Conclusion: Dsol reduced hepatic IRI after extended cold preservation and subsequent reperfusion. The protection was primarily due to the maintenance of mitochondrial function, cytoskeletal integrity, leading to limiting oxidative stress, apoptosis, and necrosis pathways.

Highlights

Cold preservation and subsequent reperfusion causes early graft dysfunction [1] and resulting poor outcome in liver transplantation [2]
We have reported the efficacy of D2O containing solution (Dsol) in a model of a prolonged cold storage and transplantation of rat hearts, shown by the inhibition of Ca2+ overload during cold preservation and by less activation of proteases thereafter [10]
Antibodies, phosphorylated-LIM kinase (LIMK)1, Glyceraldehyde 3-phosphate dehydrogenase (GAPDH), and the horseradish peroxidase-conjugated anti-rabbit IgG secondary antibody were purchased from Cell Signaling Technology (Beverly, MA, USA)

Summary

Introduction

Cold preservation and subsequent reperfusion causes early graft dysfunction [1] and resulting poor outcome in liver transplantation [2]. Cold preservation primarily causes mitochondrial dysfunction and energy depletion [3], initiating oxidative stress [4], Ca2+ overload [5], cellular swelling [6], and cytoskeletal disruption [7]. These harmful responses are interrelated and further propagated during the early phase of reperfusion (1–3 hours), and reach apoptosis and/or necrosis thereafter [8]. We have reported the efficacy of Dsol in a model of a prolonged cold storage and transplantation of rat hearts, shown by the inhibition of Ca2+ overload during cold preservation and by less activation of proteases thereafter [10]. The protection was primarily due to the maintenance of mitochondrial function, cytoskeletal integrity, leading to limiting oxidative stress, apoptosis, and necrosis pathways

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