Skip to main content
NCBI home page
Annals of Surgery logoLink to Annals of Surgery
. 1998 Jun;227(6):832–840. doi: 10.1097/00000658-199806000-00006

Reduction of hepatic ischemia/reperfusion injury by a soluble P-selectin glycoprotein ligand-1.

T S Dulkanchainun 1, J A Goss 1, D K Imagawa 1, G D Shaw 1, D M Anselmo 1, F Kaldas 1, T Wang 1, D Zhao 1, A A Busuttil 1, H Kato 1, N G Murray 1, J W Kupiec-Weglinski 1, R W Busuttil 1
PMCID: PMC1191386  PMID: 9637546

Abstract

OBJECTIVE: The authors' goal was to determine the effects of specific binding and blockade of P- and E-selectins by a soluble P-selectin glycoprotein ligand-1 (PSGL-1) in rat models of hepatic in vivo warm ischemia and ex vivo cold ischemia. The authors also sought to determine the effect of selectin blockade on isograft survival in a syngeneic rat orthotopic liver transplant model. SUMMARY BACKGROUND DATA: Ischemia/reperfusion (I/R) injury is a major factor in poor graft function after liver transplantation, which may profoundly influence early graft function and late changes. It is hypothesized that I/R injury leads to the upregulation of P-selectin, which is then rapidly translocated to endothelial cell surfaces within 5 minutes of reperfusion of the liver, initiating steps leading to tethering of polymorphonuclear neutrophil leukocytes to the vascular intima. Local production by leukocytes of interleukin-1, tumor necrosis factor-alpha, or both induces P-selectin expression on the endothelium and continues the cascade of events, which increases cell adherence and infiltration of the organ. METHODS: To examine directly the effects of selectins in a warm hepatic I/R injury model, 100 microg of PSGL-1 or saline was given through the portal vein at the time of total hepatic inflow occlusion. The effects of PSGL-1 in cold ischemia were assessed using an isolated perfused rat liver after 6 hours of 4 degrees C storage in University of Wisconsin (UW) solution, with or without the instillation of PSGL-1 before the storage. To evaluate the effect of selectin blockade on liver transplant survival, syngeneic orthotopic liver transplants were performed between inbred male Sprague-Dawley rats after 24 hours of cold ischemic storage in UW solution. A separate group of animals received two doses of 100 microg of PSGL-1 through the portal vein before storage and before reperfusion of the transplanted liver. Recipient survival was assessed at 7 days, and the Kaplan-Meier product limit estimate method was used for univariate calculations of time-dependent recipient survival events. RESULTS: In an in vivo warm rat liver ischemia model, perfusion with PSGL-1 afforded considerable protection from I/R injury, as demonstrated by decreased transaminase release, reduced histologic hepatocyte damage, and suppressed neutrophil infiltration, versus controls (p < 0.05). When cold stored livers were reperfused, PSGL-1 reduced the degree of hepatocyte transaminase release, reduced neutrophil infiltration, and decreased histologic hepatocyte damage (p < 0.05 vs. UW-only controls). On reperfusion, livers treated with PSGL-1 demonstrated increased portal vein blood flow and bile production (p < 0.05 vs. UW-only controls). In addition, 90% of the rats receiving liver isografts stored in UW solution supplemented with PSGL-1 survived 7 days versus 50% of those whose transplanted syngeneic livers had been stored in UW alone (p < 0.05). CONCLUSIONS: Selectins play an important role in I/R injury of the liver. Early modulation of the interaction between P-selectin and its ligand decreases hepatocyte injury, neutrophil adhesion, and subsequent migration in both warm and cold rat liver ischemia models. In addition, the use of PSGL-1 before ischemic storage and before transplantation prevents hepatic injury, as documented by a significant increase in liver isograft survival. These findings have important clinical ramifications: early inhibition of alloantigen-independent mechanisms during the I/R damage may influence both short- and long-term survival of liver allografts.

Full text

PDF
832

Images in this article

836Figure 1.
on p.836
836Figure 2.
on p.836

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Asa D., Raycroft L., Ma L., Aeed P. A., Kaytes P. S., Elhammer A. P., Geng J. G. The P-selectin glycoprotein ligand functions as a common human leukocyte ligand for P- and E-selectins. J Biol Chem. 1995 May 12;270(19):11662–11670. doi: 10.1074/jbc.270.19.11662. [DOI] [PubMed] [Google Scholar]
  2. Bierhuizen M. F., Fukuda M. Expression cloning of a cDNA encoding UDP-GlcNAc:Gal beta 1-3-GalNAc-R (GlcNAc to GalNAc) beta 1-6GlcNAc transferase by gene transfer into CHO cells expressing polyoma large tumor antigen. Proc Natl Acad Sci U S A. 1992 Oct 1;89(19):9326–9330. doi: 10.1073/pnas.89.19.9326. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bradford B. U., Marotto M., Lemasters J. J., Thurman R. G. New, simple models to evaluate zone-specific damage due to hypoxia in the perfused rat liver: time course and effect of nutritional state. J Pharmacol Exp Ther. 1986 Jan;236(1):263–268. [PubMed] [Google Scholar]
  4. Bradley P. P., Priebat D. A., Christensen R. D., Rothstein G. Measurement of cutaneous inflammation: estimation of neutrophil content with an enzyme marker. J Invest Dermatol. 1982 Mar;78(3):206–209. doi: 10.1111/1523-1747.ep12506462. [DOI] [PubMed] [Google Scholar]
  5. Bzeizi K. I., Jalan R., Plevris J. N., Hayes P. C. Primary graft dysfunction after liver transplantation: from pathogenesis to prevention. Liver Transpl Surg. 1997 Mar;3(2):137–148. doi: 10.1002/lt.500030206. [DOI] [PubMed] [Google Scholar]
  6. Colletti L. M., Remick D. G., Burtch G. D., Kunkel S. L., Strieter R. M., Campbell D. A., Jr Role of tumor necrosis factor-alpha in the pathophysiologic alterations after hepatic ischemia/reperfusion injury in the rat. J Clin Invest. 1990 Jun;85(6):1936–1943. doi: 10.1172/JCI114656. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Decker K. Biologically active products of stimulated liver macrophages (Kupffer cells). Eur J Biochem. 1990 Sep 11;192(2):245–261. doi: 10.1111/j.1432-1033.1990.tb19222.x. [DOI] [PubMed] [Google Scholar]
  8. Gao W., Connor H. D., Lemasters J. J., Mason R. P., Thurman R. G. Primary nonfunction of fatty livers produced by alcohol is associated with a new, antioxidant-insensitive free radical species. Transplantation. 1995 Mar 15;59(5):674–679. doi: 10.1097/00007890-199503150-00005. [DOI] [PubMed] [Google Scholar]
  9. Garcia-Criado F. J., Toledo-Pereyra L. H., Lopez-Neblina F., Phillips M. L., Paez-Rollys A., Misawa K. Role of P-selectin in total hepatic ischemia and reperfusion. J Am Coll Surg. 1995 Oct;181(4):327–334. [PubMed] [Google Scholar]
  10. Gotsch U., Jäger U., Dominis M., Vestweber D. Expression of P-selectin on endothelial cells is upregulated by LPS and TNF-alpha in vivo. Cell Adhes Commun. 1994 Apr;2(1):7–14. doi: 10.3109/15419069409014198. [DOI] [PubMed] [Google Scholar]
  11. Grober J. S., Bowen B. L., Ebling H., Athey B., Thompson C. B., Fox D. A., Stoolman L. M. Monocyte-endothelial adhesion in chronic rheumatoid arthritis. In situ detection of selectin and integrin-dependent interactions. J Clin Invest. 1993 Jun;91(6):2609–2619. doi: 10.1172/JCI116500. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Guyer D. A., Moore K. L., Lynam E. B., Schammel C. M., Rogelj S., McEver R. P., Sklar L. A. P-selectin glycoprotein ligand-1 (PSGL-1) is a ligand for L-selectin in neutrophil aggregation. Blood. 1996 Oct 1;88(7):2415–2421. [PubMed] [Google Scholar]
  13. Hamamoto I., Hossain M. A., Mori S., Okano K., Okada S., Wakabayashi H., Maeba T., Maeta H. The selectin family is responsible for microcirculatory disturbances after cold ischemia and reperfusion of the liver. Transplant Proc. 1996 Feb;28(1):68–69. [PubMed] [Google Scholar]
  14. Hamburger S. A., McEver R. P. GMP-140 mediates adhesion of stimulated platelets to neutrophils. Blood. 1990 Feb 1;75(3):550–554. [PubMed] [Google Scholar]
  15. Hayashi M., Tokunaga Y., Fujita T., Tanaka K., Yamaoka Y., Ozawa K. The effects of cold preservation on steatotic graft viability in rat liver transplantation. Transplantation. 1993 Aug;56(2):282–287. doi: 10.1097/00007890-199308000-00005. [DOI] [PubMed] [Google Scholar]
  16. Hertl M., Howard T. K., Lowell J. A., Shenoy S., Robert P., Harvey C., Strasberg S. M. Changes in liver core temperature during preservation and rewarming in human and porcine liver allografts. Liver Transpl Surg. 1996 Mar;2(2):111–117. doi: 10.1002/lt.500020205. [DOI] [PubMed] [Google Scholar]
  17. Husberg B. S., Genyk Y. S., Klintmalm G. B. A new rat model for studies of the ischemic injury after transplantation of fatty livers: improvement after postoperative administration of prostaglandin. Transplantation. 1994 Feb;57(3):457–458. doi: 10.1097/00007890-199402150-00025. [DOI] [PubMed] [Google Scholar]
  18. Jones S. M., Thurman R. G. L-arginine minimizes reperfusion injury in a low-flow, reflow model of liver perfusion. Hepatology. 1996 Jul;24(1):163–168. doi: 10.1002/hep.510240127. [DOI] [PubMed] [Google Scholar]
  19. Kamada N., Calne R. Y. Orthotopic liver transplantation in the rat. Technique using cuff for portal vein anastomosis and biliary drainage. Transplantation. 1979 Jul;28(1):47–50. [PubMed] [Google Scholar]
  20. Kaufman R. J., Davies M. V., Wasley L. C., Michnick D. Improved vectors for stable expression of foreign genes in mammalian cells by use of the untranslated leader sequence from EMC virus. Nucleic Acids Res. 1991 Aug 25;19(16):4485–4490. doi: 10.1093/nar/19.16.4485. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Kukowska-Latallo J. F., Larsen R. D., Nair R. P., Lowe J. B. A cloned human cDNA determines expression of a mouse stage-specific embryonic antigen and the Lewis blood group alpha(1,3/1,4)fucosyltransferase. Genes Dev. 1990 Aug;4(8):1288–1303. doi: 10.1101/gad.4.8.1288. [DOI] [PubMed] [Google Scholar]
  22. Lawrence M. B., Springer T. A. Leukocytes roll on a selectin at physiologic flow rates: distinction from and prerequisite for adhesion through integrins. Cell. 1991 May 31;65(5):859–873. doi: 10.1016/0092-8674(91)90393-d. [DOI] [PubMed] [Google Scholar]
  23. Lemasters J. J., Bunzendahl H., Thurman R. G. Reperfusion injury to donor livers stored for transplantation. Liver Transpl Surg. 1995 Mar;1(2):124–138. doi: 10.1002/lt.500010211. [DOI] [PubMed] [Google Scholar]
  24. Misawa K., Toledo-Pereyra L. H., Phillips M. L., Garcia-Cirado F. J., Lopez-Neblina F., Paez-Rollys A. Role of sialyl Lewis(x) in total hepatic ischemia and reperfusion. J Am Coll Surg. 1996 Mar;182(3):251–256. [PubMed] [Google Scholar]
  25. Mulligan M. S., Polley M. J., Bayer R. J., Nunn M. F., Paulson J. C., Ward P. A. Neutrophil-dependent acute lung injury. Requirement for P-selectin (GMP-140). J Clin Invest. 1992 Oct;90(4):1600–1607. doi: 10.1172/JCI116029. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Olthoff K. M., Wasef E., Seu P., Imagawa D. K., Freischlag J. A., Hart J., Busuttil R. W. PGE1 reduces injury in hepatic allografts following preservation. J Surg Res. 1991 Jun;50(6):595–601. doi: 10.1016/0022-4804(91)90048-q. [DOI] [PubMed] [Google Scholar]
  27. Palabrica T., Lobb R., Furie B. C., Aronovitz M., Benjamin C., Hsu Y. M., Sajer S. A., Furie B. Leukocyte accumulation promoting fibrin deposition is mediated in vivo by P-selectin on adherent platelets. Nature. 1992 Oct 29;359(6398):848–851. doi: 10.1038/359848a0. [DOI] [PubMed] [Google Scholar]
  28. Patel K. D., McEver R. P. Comparison of tethering and rolling of eosinophils and neutrophils through selectins and P-selectin glycoprotein ligand-1. J Immunol. 1997 Nov 1;159(9):4555–4565. [PubMed] [Google Scholar]
  29. Pinsky D. J., Naka Y., Liao H., Oz M. C., Wagner D. D., Mayadas T. N., Johnson R. C., Hynes R. O., Heath M., Lawson C. A. Hypoxia-induced exocytosis of endothelial cell Weibel-Palade bodies. A mechanism for rapid neutrophil recruitment after cardiac preservation. J Clin Invest. 1996 Jan 15;97(2):493–500. doi: 10.1172/JCI118440. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Roland C. R., Mangino M. J., Duffy B. F., Flye M. W. Lymphocyte suppression by Kupffer cells prevents portal venous tolerance induction: a study of macrophage function after intravenous gadolinium. Transplantation. 1993 May;55(5):1151–1158. doi: 10.1097/00007890-199305000-00041. [DOI] [PubMed] [Google Scholar]
  31. Roland C. R., Walp L., Stack R. M., Flye M. W. Outcome of Kupffer cell antigen presentation to a cloned murine Th1 lymphocyte depends on the inducibility of nitric oxide synthase by IFN-gamma. J Immunol. 1994 Dec 15;153(12):5453–5464. [PubMed] [Google Scholar]
  32. Sako D., Chang X. J., Barone K. M., Vachino G., White H. M., Shaw G., Veldman G. M., Bean K. M., Ahern T. J., Furie B. Expression cloning of a functional glycoprotein ligand for P-selectin. Cell. 1993 Dec 17;75(6):1179–1186. doi: 10.1016/0092-8674(93)90327-m. [DOI] [PubMed] [Google Scholar]
  33. Sako D., Comess K. M., Barone K. M., Camphausen R. T., Cumming D. A., Shaw G. D. A sulfated peptide segment at the amino terminus of PSGL-1 is critical for P-selectin binding. Cell. 1995 Oct 20;83(2):323–331. doi: 10.1016/0092-8674(95)90173-6. [DOI] [PubMed] [Google Scholar]
  34. Serizawa A., Nakamura S., Suzuki, Baba S., Nakano M. Involvement of platelet-activating factor in cytokine production and neutrophil activation after hepatic ischemia-reperfusion. Hepatology. 1996 Jun;23(6):1656–1663. doi: 10.1002/hep.510230649. [DOI] [PubMed] [Google Scholar]
  35. Suzuki S., Toledo-Pereyra L. H., Rodriguez F. J., Cejalvo D. Neutrophil infiltration as an important factor in liver ischemia and reperfusion injury. Modulating effects of FK506 and cyclosporine. Transplantation. 1993 Jun;55(6):1265–1272. doi: 10.1097/00007890-199306000-00011. [DOI] [PubMed] [Google Scholar]
  36. Takada M., Nadeau K. C., Shaw G. D., Marquette K. A., Tilney N. L. The cytokine-adhesion molecule cascade in ischemia/reperfusion injury of the rat kidney. Inhibition by a soluble P-selectin ligand. J Clin Invest. 1997 Jun 1;99(11):2682–2690. doi: 10.1172/JCI119457. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Teramoto K., Bowers J. L., Kruskal J. B., Clouse M. E. Hepatic microcirculatory changes after reperfusion in fatty and normal liver transplantation in the rat. Transplantation. 1993 Nov;56(5):1076–1082. doi: 10.1097/00007890-199311000-00005. [DOI] [PubMed] [Google Scholar]
  38. Tomita I., Sawa M., Munakata T., Tanaka K., Kasai S. The beneficial effect of dibutyryl cyclic adenosine monophosphate on warm ischemic injury of the rat liver induced by cardiac arrest. Transplantation. 1996 Jul 27;62(2):167–173. doi: 10.1097/00007890-199607270-00004. [DOI] [PubMed] [Google Scholar]
  39. Trevisani F., Colantoni A., Caraceni P., Van Thiel D. H. The use of donor fatty liver for liver transplantation: a challenge or a quagmire? J Hepatol. 1996 Jan;24(1):114–121. doi: 10.1016/s0168-8278(96)80195-4. [DOI] [PubMed] [Google Scholar]
  40. Vachino G., Chang X. J., Veldman G. M., Kumar R., Sako D., Fouser L. A., Berndt M. C., Cumming D. A. P-selectin glycoprotein ligand-1 is the major counter-receptor for P-selectin on stimulated T cells and is widely distributed in non-functional form on many lymphocytic cells. J Biol Chem. 1995 Sep 15;270(37):21966–21974. doi: 10.1074/jbc.270.37.21966. [DOI] [PubMed] [Google Scholar]
  41. Weyrich A. S., Ma X. Y., Lefer D. J., Albertine K. H., Lefer A. M. In vivo neutralization of P-selectin protects feline heart and endothelium in myocardial ischemia and reperfusion injury. J Clin Invest. 1993 Jun;91(6):2620–2629. doi: 10.1172/JCI116501. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Zhong Z., Connor H., Mason R. P., Qu W., Stachlewitz R. F., Gao W., Lemasters J. J., Thurman R. G. Destruction of Kupffer cells increases survival and reduces graft injury after transplantation of fatty livers from ethanol-treated rats. Liver Transpl Surg. 1996 Sep;2(5):383–387. doi: 10.1002/lt.500020509. [DOI] [PubMed] [Google Scholar]

Articles from Annals of Surgery are provided here courtesy of Lippincott, Williams, and Wilkins

RESOURCES