Increased Urokinase-Type Plasminogen Activator (u-PA) Levels in Graft Liver Perfusate and Decreased Single Chain u-PA Activation with Higher Levels of Aprotinin

Summary

In orthotopic liver transplantation (OLT) the graft liver is perfused with arterial blood prior to the opening of the hepatocaval anastomosis. In the present investigation we focused on the reperfusion of the graft liver in order to study the hepatic influence in the regulation of urokinase-type plasminogen activator (u-PA levels). Two different aprotinin schedules were used in 43 patients. We measured u-PA levels in the perfusate and in the corresponding systemic circulation. u-PA levels were higher in the perfusate as compared to systemic blood samples despite the dilution of the perfusate sample by the preservation fluid. This suggests u-PA secretion by the graft liver. In the presence of lower aprotinin levels signs of single-chain n-PA (scu-PA) activation was in the perfusate more prominent than systemically – a difference which was not seen in the presence of higher aprotinin levels. This seems to be an argument for the effectiveness of higher dosed aprotinin application in preventing scu-PA activation.

• References

• 1 Binnema DJ, Dooijewaard G, Turion PN C. An analysis of the activators of single-chain urokinase-type plasminogen activator (scuPA) in the dextran sulphate euglobulin fraction of normal plasma and plasmas deficient in factor XII and prekallikrein. Thromb Haemostas 1991; 65: 144-8
• 2 Booyse FM, Scheinbuks J, Hua Lin P, Traylor M, Bruce R. Isolation and interrelationships of the multiple molecular tissue-type and urokinase-type plasminogen activator forms produced by cultured human umbilical vein endothelial cells. J Biol Chem 1988; 263: 15129-38
• 3 Haucrt J, Nicoloso G, Schlcuning W-D, Bachmann F, Schapira M. Plasminogen activators in dextran sulphate activated euglobulin frac tion: a molecular analysis of factor XTT- and prekallikrein-dependent fibrinolysis. Blood 1989; 73: 994-9
• 4 Ichiiiose A, Fujikawa K, Suyama T. The activaliun of pro-uiokinase by plasma kallikrein and its inactivation by thrombin. J Biol Chem 1986; 261: 3486-9
• 5 Kluft C, Dooijewaard G, Emeis JJ. Role of the contact system in fibrinolysis. Semin Thromb Haemostas 1987; 13: 50-68
• 6 Bontempo FA, Lewis JH, Van Thiel DH, Spcro JA, Ragni MV, Butler P, Israel L, Starzl TR. The relation of preoperative coagulation findings to diagnosis, blood usage, and survival in adult liver transplantation. Transplantation 1985; 39: 532-6
• 7 Starzl TE, Demetris AJ, Van Thiel D. Liver transplantation. New Engl J Med 1984; 311: 1658-64
• 8 Dzik WH, Arkin CF, Jenkins RL, Stump DC. Fibrinolysis during liver transplantation in humans: role of tissue-type plasminogen activator. Blood 1988; 71: 1090-5
• 9 Himmelreich G, Kierzek B, Neuhaus P, Slama K-J, Riess H. Coagulation changes and the influence of the early perfusate in the course of orthotopic liver transplantation (OLT) when aprotinin is used intraoperatively. Blood Coagulation and Fibrinolysis 1991; 2: 51-9
• 10 Palareti G, De Rosa V, Fortunato G, Grauso F, Legnani C, Maccaferri M, Poggi M, Bianchini B, Bellusci R, Franceschelli N, Coccheri S. Control of hemostasis during orthotopic liver transplantation. Fibrinolysis 1988; (02) suppl (Suppl. 03) 61-6
• 11 Porte RJ, Bontempo FA, Knot EA R, Lewis JH, Kang YG, Starzl TE. Systemic effects of tissue plasminogen activator-associated fibrinolysis and its relation to thrombin generation in orthotopic liver transplantation. Transplantation 1989; 47: 978-84
• 12 Himmelreich G, Dooijewaard G, Breinl P, Bechstein WO, Neuhaus P, Kluft C, Riess H. Evolution of urokinase-type plasminogen activator (u-PA) and tissue-type plasminogen activator (t-PA) in orthotopic liver transplantation (OLT). Thromb Haemostas 1993; 69: 56-9
• 13 Fritz H, Wunderer G, Jochum M. Biochemistry and applications of aprotinin, the kallikrein inhibitor from bovine organs. Arzneim Forsch/Drug Res 1983; 33: 479-94
• 14 Verstraete M. Clinical applications of inhibitors of fibrinolysis. Drugs 1985; 29: 236-61
• 15 Grosse H, Lobbes W, Frambach M, Ringe B, Barthels M. Influence of high dose aprotinin on hemostasis and blood loss in orthotopic liver transplantation (OLT). Semin Thromb Haemostas 1993; 19: 302-5
• 16 Mallett SV, Cox D, Burroughs AK, Rolles K. Aprotinin and reduction of blood loss and transfusion requirements in orthotopic liver transplantation. Lancet 1990; 336: 886
• 17 Neuhaus P, Bechstein WO, Hopf U, Blumhardt G, Steffen R. Indikationen und aktuelle Entwicklung der Lebertransplantation. Leber Magen Darm 1989; 19: 289-308
• 18 Neuhaus P, Blumhardt G, Bechstein WO, Steffen R, Keck H. Side-to-side anastomosis of the common bile duct is the method of choice for biliary tract reconstruction after liver transplantation. Transplant Proc 1990; 22: 1571
• 19 Binnema DJ, Van Iersel JJ L, Dooijewaard G. Quantitation of urokinase antigen in plasma and culture media by use of an ELISA. Thromb Res 1986; 43: 569-77
• 20 van Hinsbergh VW M, van den Berg EA, Fiers W, Dooijewaard G. Tumor necrosis factor induces the production of urokinase-type plasminogen activator by human endothelial cells. Blood 1990; 75: 1991-8
• 21 Vcrhcijcn JH, Mullaart E, Chang GT G, Kluft C, Wijngaards G. A simple, sensitive spectrophotomctric assay for extrinsic (tissuc-typc) plasminogen activator applicable to measurements in plasma. Thromb Haemostas 1982; 48: 266-9
• 22 Kircliheimci JC, Hubei K, Polleiauei P, Binder BR. Urokinase antigen in plasma of patients with liver cirrhosis and hepatoma. Thromb Haemostas 1985; 54: 617-8
• 23 Himmelreich M, Neuhaus P, Bechstein WO, Slama K-J, Jochum M, Riess H. Different aprotinin applications influencing hemosLulie ehanges in orlhotopic liver transplantation. Transplantation 1992; 53: 132-6
• 24 Bachmann F. Fibrinolysis. In: Thrombosis and Haemostasis. International Society on Thrombosis and Haemostasis. Verstraete M, Ver mylen J, Lijnen HR, Arnout J. (eds) Leuven University Press; Leuven, Belgium: 1987: 227-63
• 25 Kluft C. Levels of plasminogen activators in human plasma: new methods to study the intrinsic and extrinsic activators. In: Progress in chemical fibrinolysis and thrombolysis. Davidson JF, Rowen RM, Samama MM, Desnoyers PC. (eds) Raven Press; New York, NY: 1978. (; Vol 3). 141-64
• 26 Lie TS, Seger R, Hong GS, Preissinger H, Ogawa K. Protective effect of aprotinin on ischemic hepatocellular damage. Transplantation 1989; 48: 396-9
• 27 Riess H, Jochum M, Machleidt W, Himmelreich G, Roissaint R, Steffen R. Possible role of extracellularly released phagocyte proteinases in the coagulation disorder during liver transplantation. Transplantation 1991; 52: 482-90
• 28 Kobayasti H, Schmitt M, Goretzki L, Chucholowski N, Calvete J, Kramer M, Giinzler WA, Janicke F, Graeff H. Cathepsin B efficiently activates the soluble and the tumor cell receptor-bound form of the proenzyme urokinase-type plasminogen activator (pro-u-PA). J Biol Chem 1991; 266: 5147-52