Post-reperfusion hydrogen gas treatment ameliorates ischemia reperfusion injury in rat livers from donors after cardiac death: a preliminary study

Abstract

Background and purpose

We reported previously that hydrogen gas (H2) reduced hepatic ischemia and reperfusion injury (IRI) after prolonged cold storage (CS) of livers retrieved from heart-beating donors. The present study was designed to assess whether H2 reduced hepatic IRI during donation of a cardiac death (DCD) graft with subsequent CS.

Methods

Rat livers were harvested after 30-min cardiac arrest and stored for 4 h in University of Wisconsin solution. The graft was reperfused with oxygenated buffer, with or without H2 (H2 or NT groups, respectively), at 37° for 90 min on isolated perfused rat liver apparatus.

Results

In the NT group, liver enzyme leakage, apoptosis, necrosis, energy depletion, redox status, impaired microcirculation, and bile production were indicative of severe IRI, whereas in the H2 group these impairments were significantly suppressed. The phosphorylation of cytoplasmic MKK4 and JNK were enhanced in the NT group and suppressed in the H2 group. NFkB-p65 and c-Fos in the nucleus were unexpectedly unchanged by IRI regardless of H2 treatment, indicating the absence of inflammation in this model.

Conclusion

H2 was observed to ameliorate IRI in the DCD liver by maintaining microcirculation, mitochondrial functions, and redox status, as well as suppressing the cytoplasmic MKK4–JNK-mediated cellular death pathway.

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Abbreviations

ATP:

Adenosine triphosphate

ALT:

Alanine aminotransferase

AST:

Aspartate aminotransferase

CPA:

Cardiopulmonary arrest

DCD:

Donation after cardiac death

DBD:

Donation after brain death

ECD:

Expanded criteria donor

GSSG:

Oxidized glutathione

GSH:

Reduced glutathione

HPFs:

High power fields

H2 :

Hydrogen gas

4-HNE:

4-Hydroxy-2-nonenal

IRI:

Ischemia and reperfusion injury

IPRL:

Isolated perfused rat liver

JNK:

c-jun N-terminal kinase

KHB:

Krebs–Henseleit bicarbonate buffer

LDH:

Lactate dehydrogenase

LPO:

Lipid peroxidation

MDA:

Malondialdehyde

MKK4:

Mitogen-activated protein kinase 4

NADPH:

Nicotinamide adenine dinucleotide phosphate

NMP:

Normothermic machine perfusion

OCR:

Oxygen consumption rate

PNF:

Primary non-function

PVP:

Portal vein pressure

PVR:

Portal vein resistance

ROS:

Reactive oxygen species

SCD:

Standard criteria donor

SDS-PAGE:

SDS polyacrylamide gel electrophoresis

TUNEL:

Terminal deoxynucleotidyl transferase

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Acknowledgements

We thank Mr. Masatoshi Horigome for the animal care, Ms. Sayaka Miyoshi for technical assistance, and the staff of the Department of Gastroenterological Surgery I, Hokkaido University Graduate School of Medicine, for their kind cooperation.

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Correspondence to Moto Fukai.

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Ishikawa, T., Shimada, S., Fukai, M. et al. Post-reperfusion hydrogen gas treatment ameliorates ischemia reperfusion injury in rat livers from donors after cardiac death: a preliminary study. Surg Today 48, 1081–1088 (2018). https://doi.org/10.1007/s00595-018-1693-0

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Keywords

  • Hydrogen gas
  • Donation after cardiac death
  • Liver
  • Rat
  • Ischemia reperfusion