Amino Acids

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Glycine protects partial liver grafts from Kupffer cell-dependent ischemia–reperfusion injury without negative effect on regeneration

  • Mohammed Al-Saeedi
  • Rui Liang
  • Daniel P. Schultze
  • Arash Nickkholgh
  • Ingrid Herr
  • Markus Zorn
  • Peter SchemmerEmail author
Original Article


Donor preconditioning with glycine prevents Kupffer cell-dependent reperfusion injury to liver grafts. Partial liver grafts need to regenerate and grow in size after transplantation; however, glycine inactivates Kupffer cells, which are important for hepatic regeneration. Thus, this study was designed to evaluate the impact of donor preconditioning with glycine after partial liver transplantation (pLTx). PLTx was performed in 28 female Sprague–Dawley rats. Glycine (1.5 ml, 300 mM; i.v.) was given to 14 live donors before organ procurement. Liver enzymes and histology were investigated 8 h after reperfusion to index liver injury and leukocyte infiltration. Hepatic microperfusion and leukocyte–endothelium interaction were assessed using the in vivo fluorescence microscopy method. Ki-67 and TNF-α were detected by immunohistochemistry for regeneration and Kupffer cell activation. Glycine significantly increased survival from 0% in controls to 40%, while both liver enzyme levels and necrosis were decreased to about 50% of controls (p < 0.05). Sinusoidal blood flow increased by 40–80%, while leukocyte–endothelium interaction decreased to 30% of control values (p < 0.05). While Kupffer cell-derived TNF-α decreased to 70% of controls, there was no difference between groups in Ki-67 expression. Data presented here clearly demonstrate that glycine protects partial liver grafts from reperfusion injury without effects on regeneration.


Partial liver transplantation Regeneration Ischemia-/reperfusion injury Glycine TNF-α Ki-67 Kupffer cells 


Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical approval

All applicable international, national and/or institutional guidelines for the care and use of animals were followed.


  1. Al-Saeedi M et al (2018a) Neutralization of CD95 ligand protects the liver against ischemia-reperfusion injury and prevents acute liver failure. Cell Death Dis 9(2):132Google Scholar
  2. Al-Saeedi M et al (2018b) Glycine protects the liver from reperfusion injury following pneumoperitoneum. Eur Surg Res 59(1–2):91–99Google Scholar
  3. Benko T et al (2010) Glycine pretreatment ameliorates liver injury after partial hepatectomy in the rat. J Invest Surg 23(1):12–20Google Scholar
  4. Bergmeyer HU, Scheibe P, Wahlefeld AW (1978) Optimization of methods for aspartate aminotransferase and alanine aminotransferase. Clin Chem 24(1):58–73Google Scholar
  5. Byun SH, Yang HS, Kim JH (2016) Liver graft hyperperfusion in the early postoperative period promotes hepatic regeneration 2 weeks after living donor liver transplantation: a prospective observational cohort study. Medicine (Baltimore) 95(46):e5404Google Scholar
  6. Conzelmann L et al (2002) Orthotopic liver transplantation in knockout mice: is TNFalpha involved in early graft injury and regeneration? Transplant Proc 34(6):2299–2300Google Scholar
  7. Conzelmann LO et al (2003) Reduced-size liver transplantation in the mouse. Transplantation 76(3):496–501Google Scholar
  8. Czigany Z et al (2015) Improving research practice in rat orthotopic and partial orthotopic liver transplantation: a review, recommendation, and publication guide. Eur Surg Res 55(1–2):119–138Google Scholar
  9. den Butter G et al (1993) Effect of glycine in dog and rat liver transplantation. Transplantation 56(4):817–822Google Scholar
  10. Enomoto N et al (1999) Development of a new, simple rat model of early alcohol-induced liver injury based on sensitization of Kupffer cells. Hepatology 29(6):1680–1689Google Scholar
  11. Fausto N (2000) Liver regeneration. J Hepatol 32(1 Suppl):19–31Google Scholar
  12. Gao W, Lemasters JJ, Thurman RG (1993) Development of a new method for hepatic rearterialization in rat orthotopic liver transplantation. Reduction of liver injury and improvement of surgical outcome by arterialization. Transplantation 56(1):19–24Google Scholar
  13. Genoves P et al (2014) Pentoxifylline in liver ischemia and reperfusion. J Invest Surg 27(2):114–124Google Scholar
  14. Ikejima K et al (1997) Kupffer cells contain a glycine-gated chloride channel. Am J Physiol 272(6 Pt 1):G1581–G1586Google Scholar
  15. Inoue A et al (2017) Regulation of matrix metalloproteinase-1 and alpha-smooth muscle actin expression by interleukin-1 alpha and tumour necrosis factor alpha in hepatic stellate cells. Cytotechnology 69(3):461–468Google Scholar
  16. Ito K et al (2008) Effect of non-essential amino acid glycine administration on the liver regeneration of partially hepatectomized rats with hepatic ischemia/reperfusion injury. Clin Nutr 27(5):773–780Google Scholar
  17. Kasprzak A et al (2004) Expression of cytokines (TNF-alpha, IL-1alpha, and IL-2) in chronic hepatitis C: comparative hybridocytochemical and immunocytochemical study in children and adult patients. J Histochem Cytochem 52(1):29–38Google Scholar
  18. Liu G et al (2017) Splenectomy after partial hepatectomy accelerates liver regeneration in mice by promoting tight junction formation via polarity protein Par 3-aPKC. Life Sci 192:91–98Google Scholar
  19. Lopez BG et al (2011) Characterization of Kupffer cells in livers of developing mice. Comp Hepatol 10(1):2Google Scholar
  20. Luntz SP et al (2005) HEGPOL: randomized, placebo controlled, multicenter, double-blind clinical trial to investigate hepatoprotective effects of glycine in the postoperative phase of liver transplantation [ISRCTN69350312]. BMC Surg 5:18Google Scholar
  21. Mehrabi A et al (2004) Long-term results of paediatric kidney transplantation at the University of Heidelberg: a 35 year single-centre experience. Nephrol Dial Transplant 19(Suppl 4):iv69–iv74Google Scholar
  22. Muller SA et al (2007) Partial liver transplantation-living donor liver transplantation and split liver transplantation. Nephrol Dial Transplant 22(Suppl 8):viii13–viii22Google Scholar
  23. Nichols JC et al (1994) Inhibition of nonlysosomal calcium-dependent proteolysis by glycine during anoxic injury of rat hepatocytes. Gastroenterology 106(1):168–176Google Scholar
  24. Okamura Y et al (2018) Coexistence of bilirubin ≥ 10 mg/dL and prothrombin time-international normalized ratio ≥ 1.6 on Day 7: a strong predictor of early graft loss after living donor liver transplantation. Transplantation 102(3):440–447Google Scholar
  25. Petrat F et al (2012) Glycine, a simple physiological compound protecting by yet puzzling mechanism(s) against ischaemia–reperfusion injury: current knowledge. Br J Pharmacol 165(7):2059–2072Google Scholar
  26. Pomposelli JJ et al (2016) Patterns of early allograft dysfunction in adult live donor liver transplantation: the A2ALL experience. Transplantation 100(7):1490–1499Google Scholar
  27. Post S et al (1993) Differential impact of Carolina rinse and University of Wisconsin solutions on microcirculation, leukocyte adhesion, Kupffer cell activity and biliary excretion after liver transplantation. Hepatology 18(6):1490–1497Google Scholar
  28. Sauer P et al (2004) Living-donor liver transplantation: evaluation of donor and recipient. Nephrol Dial Transplant Suppl 4:iv11–iv15Google Scholar
  29. Schemmer P et al (1998) Gentle in situ liver manipulation during organ harvest decreases survival after rat liver transplantation: role of Kupffer cells. Transplantation 65(8):1015–1020Google Scholar
  30. Schemmer P et al (1999) Intravenous glycine improves survival in rat liver transplantation. Am J Physiol 276(4 Pt 1):G924–G932Google Scholar
  31. Schemmer P et al (2001) Glycine reduces reperfusion injury in human liver transplantation: our first patients. Transplant Proc 33(7–8):3750–3752Google Scholar
  32. Schemmer P et al (2002) Extended experience with glycine for prevention of reperfusion injury after human liver transplantation. Transplant Proc 34(6):2307–2309Google Scholar
  33. Schemmer P et al (2005a) Living related liver transplantation: the ultimate technique to expand the donor pool? Transplantation 80(1 Suppl):S138–S141Google Scholar
  34. Schemmer P et al (2005b) Taurine improves graft survival after experimental liver transplantation. Liver Transpl 11(8):950–959Google Scholar
  35. Seabra V, Stachlewitz RF, Thurman RG (1998) Taurine blunts LPS-induced increases in intracellular calcium and TNF-alpha production by Kupffer cells. J Leukoc Biol 64(5):615–621Google Scholar
  36. Taub R, Greenbaum LE, Peng Y (1999) Transcriptional regulatory signals define cytokine-dependent and -independent pathways in liver regeneration. Semin Liver Dis 19(2):117–127Google Scholar
  37. Thurman RG, Bunzendahl H, Lemasters JJ (1993) Role of sinusoidal lining cells in hepatic reperfusion injury following cold storage and transplantation. Semin Liver Dis 13(1):93–100Google Scholar
  38. Toniutto P et al (2017) Current challenges and future directions for liver transplantation. Liver Int 37(3):317–327Google Scholar
  39. Uhlmann S, Uhlmann D, Spiegel HU (1999) Evaluation of hepatic microcirculation by in vivo microscopy. J Invest Surg 12(4):179–193Google Scholar
  40. Wheeler MD, Thurman RG (1999) Production of superoxide and TNF-alpha from alveolar macrophages is blunted by glycine. Am J Physiol 277(5):L952–L959Google Scholar
  41. Wheeler MD et al (1999) Glycine: a new anti-inflammatory immunonutrient. Cell Mol Life Sci 56(9–10):843–856Google Scholar
  42. Zhong Z et al (1996) Destruction of Kupffer cells increases survival and reduces graft injury after transplantation of fatty livers from ethanol-treated rats. Liver Transpl Surg 2(5):383–387Google Scholar
  43. Zhong Z et al (1999) Glycine improves survival after hemorrhagic shock in the rat. Shock 12(1):54–62Google Scholar
  44. Zhong Z et al (2003) L-Glycine: a novel antiinflammatory, immunomodulatory, and cytoprotective agent. Curr Opin Clin Nutr Metab Care 6(2):229–240Google Scholar

Copyright information

© Springer-Verlag GmbH Austria, part of Springer Nature 2019

Authors and Affiliations

  • Mohammed Al-Saeedi
    • 1
  • Rui Liang
    • 2
  • Daniel P. Schultze
    • 1
  • Arash Nickkholgh
    • 1
  • Ingrid Herr
    • 1
  • Markus Zorn
    • 3
  • Peter Schemmer
    • 4
    • 5
    Email author
  1. 1.Department of General, Visceral, and Transplant SurgeryHeidelberg University HospitalHeidelbergGermany
  2. 2.Department of the Second Affiliated Hospital DalianDalian Medical UniversityDalianPeople’s Republic of China
  3. 3.Department of Gastroenterology, Intoxications, and Infectious DiseasesHeidelberg University HospitalHeidelbergGermany
  4. 4.Department of General, Visceral, and Transplant Surgery, University Hospital GrazMedical University of GrazGrazAustria
  5. 5.Transplant Center Graz, University Hospital GrazMedical University of GrazGrazAustria

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