[D-Ala 2, D-Leu 5] Enkephalin Improves Liver Preservation During Normothermic Ex Vivo Perfusion

J Surg Res. 2019 Sep;241:323-335. doi: 10.1016/j.jss.2019.04.010. Epub 2019 May 6.


Background: Meeting the metabolic demands of donor livers using normothermic ex vivo liver perfusion (NEVLP) preservation technology is challenging. The delta opioid agonist [D-Ala2, D-Leu5] enkephalin (DADLE) has been reported to decrease the metabolic demand in models of ischemia and cold preservation. We evaluated the therapeutic potential of DADLE by investigating its ability to protect against oxidative stress and hepatic injury during normothermic perfusion.

Materials and methods: Primary rat hepatocytes were used in an in vitro model of oxidative stress to determine the minimum dose of DADLE needed to induce protection and the mechanisms associated with protection. NEVLP was then used to induce injury in rat livers and determine the effectiveness of DADLE in preventing liver injury.

Results: In hepatocytes, DADLE was protective against oxidative stress and led to a decrease in phosphorylation of JNK and p38. Naltrindole, a δ-opioid receptor antagonist, blocked this effect. DADLE also activated the PI3K/Akt signaling pathway, and PI3K/Akt inhibition decreased the protective effects of DADLE treatment. In addition, DADLE treatment during NEVLP resulted in lower perfusate alanine aminotransferase and tissue malondialdehyde and better tissue adenosine triphosphate and glutathione. Furthermore, perfusion with DADLE compared with perfusate alone preserved tissue architecture.

Conclusions: DADLE confers protection against oxidative stress in hepatocytes and during NEVLP. These data suggest that the mechanism of protection involved the prevention of mitochondrial dysfunction by opioid receptor signaling and subsequent increased expression of prosurvival/antiapoptotic signaling pathways. Altogether, data suggest that opioid receptor agonism may serve as therapeutic target for improved liver protection during NEVLP.

Keywords: D-Ala2,D-Leu5 enkephalin (DADLE); Liver; Naltrindole; Normothermic ex-vivo liver perfusion; Transplantation.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Allografts / drug effects*
  • Allografts / metabolism
  • Allografts / pathology
  • Animals
  • Disease Models, Animal
  • Enkephalin, Leucine-2-Alanine / pharmacology*
  • Hepatocytes
  • Humans
  • Liver / drug effects*
  • Liver / metabolism
  • Liver / pathology
  • Male
  • Mitochondria / drug effects
  • Mitochondria / metabolism
  • Organ Preservation Solutions / pharmacology*
  • Oxidative Stress / drug effects
  • Perfusion / adverse effects
  • Perfusion / methods
  • Primary Cell Culture
  • Rats
  • Receptors, Opioid, delta / metabolism
  • Reperfusion Injury / etiology
  • Reperfusion Injury / pathology
  • Reperfusion Injury / prevention & control*
  • Tissue and Organ Harvesting / adverse effects
  • Tissue and Organ Harvesting / methods


  • Organ Preservation Solutions
  • Receptors, Opioid, delta
  • Enkephalin, Leucine-2-Alanine