Journal of Zhejiang University SCIENCE B

, Volume 8, Issue 5, pp 289–295 | Cite as

A novel way of liver preservation improves rat liver viability upon reperfusion

  • Kebis Anton 
  • Kukan Marián 
  • Grančič Peter 
  • Jakubovský Ján 
Science Letters

Abstract

Background/aim

Currently, the liver is cold-preserved at 0∼4 °C for experimental and clinical purposes. Here, we investigated whether milder hypothermia during the initial phase of the preservation period was beneficial for liver viability upon reperfusion.

Methods

In the first set of experiments, rat livers were preserved either conventionally in clinically used histidine-trypthopan-ketoglutarate (HTK) solution (Group A: 45 min and Group B: 24 h) or by slow cooling HTK solution (from 13 °C to 3 °C) during the initial 45 min of preservation (Group C: 24 h). In the second set of experiments, additional groups of livers were evaluated: Group BB—preservation according to Group B and Group CC—preservation according to Group C. Further, some livers were preserved at 13 °C for 24 h. Livers were then reperfused using a blood-free perfusion model.

Results

Bile production was approximately 2-fold greater in Group C compared to Group B. Alanine transaminase (ALT) and aspartate transaminase (AST) release into perfusate were 2∼3-fold higher in Group B compared to Group C. No significant differences were found in ALT and AST release between Group C and Group A. Livers in Group CC compared to Group BB exhibited significantly lower portal resistance, greater oxygen consumption and bromosulfophthalein excretion into bile and lower lactate dehydrogenase (LDH) release into perfusate. Histological evaluation of tissue sections in Group BB showed parenchymal dystrophy of hepatocytes, while dystrophy of hepatocytes was absent in Group CC. Livers preserved at 13 °C for 24 h exhibited severe ischemic injury.

Conclusion

These results suggest that the conventional way of liver preservation is not suitable at least for rat livers and that slow cooling of HTK solution during the initial phase of cold storage can improve liver viability during reperfusion.

Key words

Rat Hepatic graft Cold ischemia Liver protection Histidine-trypthopan-ketoglutarate solution (HTK) 

CLC number

R322.4+

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. Borghi-Scoazec, G., Scoazec, J.Y., Durand, F., Bernuau, J., Belghiti, J., Feldmann, G., Henin, D., Degott, C., 1997. Apoptosis after ischemia-reperfusion in human liver allografts. Liver Transplantation, 3(4):407–415. [doi:10.1002/lt.500030408]CrossRefGoogle Scholar
  2. Bowers, B.A., Branum, G.D., Rotolo, F.S., Watters, C.R., Meyers, W.C., 1987. Bile flow—an index of ischemic injury. J. Surg. Res., 42(5):565–569. [doi:10.1016/00224804(87)90033-3]PubMedCrossRefGoogle Scholar
  3. Drews, G., Deckert, F., Witzigmann, H., Hauss, J., Spiegel, H.U., 1998. Effect of rapid cooling on microperfusion of donor livers. Zentralbl. Chir., 123(3):280–284 (in German).PubMedGoogle Scholar
  4. Fujita, S., Hamamoto, I., Nakamura, K., Tanaka, K., Ozawa, K., 1993. Isolated perfusion of rat livers: effect of temperature on O2 consumption, enzyme release, energy store, and morphology. Nippon Geka Hokan., 62(2):58–70.PubMedGoogle Scholar
  5. Hertl, M., Chartrand, P.B., West, D.D., Harvey, P.R., Strasberg, S.M., 1994. The effects of hepatic preservation at 0 °C compared to 5 °C: influence of antiproteases and periodic flushing. Cryobiology, 31(5):434–440. [doi:10.1006/cryo.1994.1053]PubMedCrossRefGoogle Scholar
  6. Hertl, M., Howard, T.K., Lowell, J.A., Shenoy, S., Robert, P., Harvey, C., Strasberg, S.M., 1996. Changes in liver core temperature during preservation and rewarming in human and porcine liver allografts. Liver Transplantation, 2(2):111–117. [doi:10.1002/lt.500020205]CrossRefGoogle Scholar
  7. Kebis, A., Zvrškovec, J., 2002. Preservation/Perfusion Chamber. Patent of Slovak Republic 3151 U, A 61 G 10/02.Google Scholar
  8. Kebis, A., Kukan, M., 2006. Effect of conductive heat transfer on cold ischemia-reperfusion injury of isolated rat liver. Čes. a Slov. Gastroent. a Hepatol., 60(1):36–42 (in Slovak).Google Scholar
  9. Kebis, A., Lutterová, M., Kukan, M., Kuba, D., 2004. Protection of hypothermic rat liver against gentle manipulation injury. Československá Fyziologie, 53(4):136–141 (in Slovak).PubMedGoogle Scholar
  10. Kukan, M., 1999. The Isolated Perfused Liver as a Tool in Drug Metabolism Studies. In: Woolf, T.F. (Ed.), Handbook of Drug Metabolism. Marcel Dekker, New York, p.425–442.Google Scholar
  11. Kukan, M., Haddad, P.S., 2001. Role of hepatocytes and bile duct cells in preservation-reperfusion injury of liver grafts. Liver Transplantation, 7(5):381–400. [doi:10.1053/jlts.2001.23913]PubMedCrossRefGoogle Scholar
  12. Lemasters, J.J., Thurman, R.G., 1997. Reperfusion injury after liver preservation for transplantation. Annu. Rev. Pharmacol. Toxicol., 37(1):327–338. [doi:10.1146/annurev.pharmtox.37.1.327]PubMedCrossRefGoogle Scholar
  13. Lutterová, M., Kukan, M., Vajdová, K., Kuba, D., Mišlanová, C., Kebis, A., Danninger, F., Mráz, P., 2001. Protection of the rat liver against rewarming ischemic injury by University of Wisconsin solution. Langenbecks Arch. Surg., 386(1):31–37. [doi:10.1007/s004230000179]PubMedCrossRefGoogle Scholar
  14. Sumimoto, K., Inagaki, K., Yamada, K., Kawasaki, T., Dohi, K., 1988. Reliable indices for the determination of viability of grafted liver immediately after orthotopic transplantation. Bile flow rate and cellular adenosine triphosphate level. Transplantation, 46(4):506–509. [doi:10.1097/00007890-198810000-00007]PubMedCrossRefGoogle Scholar
  15. Yoshida, K., Matsui, Z., Wei, T., Kaibori, M., Kwon, A.H., Yamane, A., Kamiyama, Y., 1999. A novel conception for liver preservation at a temperature just above freezing point. J. Surg. Res., 81(2):216–223. [doi:10.1006/jsre.1998.5505]PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2007

Authors and Affiliations

  • Kebis Anton 
    • 1
  • Kukan Marián 
    • 1
  • Grančič Peter 
    • 1
  • Jakubovský Ján 
    • 2
  1. 1.Laboratory of Perfused Organs, Slovak Center for Organ TransplantationSlovak Medical UniversityBratislavaSlovakia
  2. 2.Department of Pathology, School of MedicineComenius UniversityBratislavaSlovakia

Personalised recommendations