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Thromb Haemost 1982; 47(02): 166-172
DOI: 10.1055/s-0038-1657155
Original Article
Schattauer GmbH Stuttgart

A Comparison of Plasminogen Activators Derived from Rat Plasma, Primary Rat Hepatocytes and Isolated Perfused Rat Liver

Yoav Sharoni
The Department of Pharmacology, Wellcome Research Laboratories, Research Triangle Park, North Carolina, U.S.A.
,
Maria C Topal
The Department of Pharmacology, Wellcome Research Laboratories, Research Triangle Park, North Carolina, U.S.A.
,
Patricia R Tuttle
The Department of Pharmacology, Wellcome Research Laboratories, Research Triangle Park, North Carolina, U.S.A.
,
Henry Berger Jr
The Department of Pharmacology, Wellcome Research Laboratories, Research Triangle Park, North Carolina, U.S.A.
› Author Affiliations
Further Information

Publication History

Received 06 October 1981

Accepted 03 March 1982

Publication Date:
13 July 2018 (online)

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Summary

Of the two cell types it was possible to culture from the dissociated rat liver, hepatocytes and Kupffer cells, only the former were fibrinolytically active. Rat hepatocytes during the first 24 hr in culture secreted two plasminogen activators with molecular weights identical to those found in rat plasma, an 80,000-dalton form (PA-80) and a 45,000-dalton form (PA-45). Partially purified preparations of plasminogen activators from both sources were subjected to isoelectric focusing (IEF) to compare characteristics further. There were three distinct peaks of PA-45 in each preparation with isoelectric points of 7.1, 7.2 and 7.4; all electrophoretic forms had the same low affinity to fibrin. PA-80 from both sources displayed similar IEF profiles with forms ranging from pH values of 7 to 8, all with the same high affinity to fibrin. The major form of PA-80 in the plasma preparation had an isoelectric point of 7.9 whereas that in the hepatocyte preparation had an isoelectric point of 7.6. The isolated perfused rat liver was also shown to produce both PA-80 and PA-45 emphasizing the physiological relevance of the findings with hepatocytes. It is concluded that in the rat hepatocytes contribute to the plasma profile with regard to the plasminogen activator content.

Key words

Plasminogen activators - Plasma - Hepatocytes - Isolated liver perfusion - Affinity to fibrin
 

  • References

  • 1 Saito H, Hamilton SM, Tavill AS, Louis L, Ratnoff OD. Production and release of plasminogen by isolated perfused rat liver. Proc Natl Acad Sci USA 1980; 77: 6837-6840
    CrossrefPubMedGoogle Scholar
  • 2 Bohmfalk JF, Fuller GM. Plasminogen is synthesized by primary cultures of rat hepatocytes. Science 1980; 209: 408-410
    CrossrefPubMedGoogle Scholar
  • 3 Raum D, Marcus D, Alper CA, Levey R, Taylor PD, Strazl TE. Synthesis of human plasminogen by the liver. Science 1980; 208: 1036-1037
    CrossrefPubMedGoogle Scholar
  • 4 Todd AS. The histological localization of plasminogen activator. J. Pathol Bacteriol 1959; 78: 281-283
    CrossrefPubMedGoogle Scholar
  • 5 Aoki N, von Kaulla KN. Dissimilarity of human vascular plasminogen activator and human urokinase. J Lab Cin Med 1971; 78: 354-362
    PubMedGoogle Scholar
  • 6 Astrup T, Thorsen S. The physiology of fibrinolysis. Med Clin North Am 1972; 56: 153-162
    CrossrefPubMedGoogle Scholar
  • 7 Levin EG, Loskutoff DJ. Comparative studies of the fibrinolytic activity of cultured vascular cells. Thromb Res 1979; 15: 869-878
    CrossrefPubMedGoogle Scholar
  • 8 Rijken DC, Wijngaards G, Welbergen J. Relationship between tissue plasminogen activator and the activators in blood and vascular wall. Thromb Res 1980; 18: 815-830
    CrossrefPubMedGoogle Scholar
  • 9 Kwaan HC, MacFadzean AJ. On plasma fibrinolytic activity induced by ischemia. Clin Sci 1956; 15: 245-257
    PubMedGoogle Scholar
  • 10 Nilsson IM, Pandolfi M. Fibrinolysis, response of the vascular wall. Thrombos Diathes Haemorrh (Suppl. 00) 1970; 40: 231-242
    PubMedGoogle Scholar
  • 11 Ogston D, Bennett B, Mackie M. Properties of a partially purified preparation of a circulating plasminogen activator. Thromb Res 1976; 8: 275-284
    CrossrefPubMedGoogle Scholar
  • 12 Radcliffe R, Heinze T. Isolation of plasminogen activator from human plasma by chromatography on lysine-Sepharose. Arch Biochem Biophys 1978; 189: 185-194
    CrossrefPubMedGoogle Scholar
  • 13 Laishes BA, Roberts E, Burrows C. Fibrinolytic activity of adult rat liver cells in primary culture and inhibition by glucocorticoids. Biochem Biophys Res Commun 1976; 72: 462-471
    CrossrefPubMedGoogle Scholar
  • 14 Williams GM, Bermudez E, San RHC, Goldblatt PJ, Laspia MF. Rat hepatocyte primary cultures IV. Maintenance in defined medium and the role of production of plasminogen activator and other proteases. In Vitro 1978; 14: 824-837
    CrossrefPubMedGoogle Scholar
  • 15 Miller LL. Technique of isolated rat liver perfusion. In: Isolated Liver Perfusion and Its Applications 1973. 11-52 Bartosek I, Guaitani A, Miller LL. (Eds) Raven Press; New York: 1973
    Google Scholar
  • 16 Frohlich J, Hansen W, Scholz R. Gluconeogenesis in rat livers perfused with nonrecirculating Krebs-Henseleit buffer. In: Isolated Liver Perfusion and Its Applications 1973. 205-210 Bartosek I, Guaitani A, Miller LL. (Eds) Raven Press; New York: 1973
    Google Scholar
  • 17 Williams GM, Bermudez E, Scaramuzzino D. Rat heptocyte primary cell cultures III. Improved dissociation and attachment techniques and the enhancement of survival by culture medium. In Vitro 1977; 13: 809-817
    CrossrefPubMedGoogle Scholar
  • 18 Berg T, Boman D. Distribution of lysosomal enzymes between parenchymal and Kupffer cells of rat liver. Biochim Biophys Acta 1973; 321: 585-596
    CrossrefPubMedGoogle Scholar
  • 19 Seglen PO. Preparation of rat liver cells III. Enzymatic requirements for tissue dispersion. Exp Cell Res 1973; 82: 391-398
    CrossrefPubMedGoogle Scholar
  • 20 Folkman J, Haudenschild CC, Zetter BR. Long-term culture of capillary endothelial cells. Proc Natl Acad Sci USA 1979; 76: 5217-5221
    CrossrefPubMedGoogle Scholar
  • 21 Michalopoulos G, Pitot HC. Primary culture of parenchymal liver cells on collagen membranes. Exp Cell Res 1975; 94: 70-78
    CrossrefPubMedGoogle Scholar
  • 22 Munthe-Kaas AC, Berg T, Seglen PO, Seljelid R. Mass isolation and culture of rat Kupffer cells. J Exp Med 1975; 141: 1-10
    CrossrefPubMedGoogle Scholar
  • 23 Jones P, Benedict W, Strickland S, Reich E. Fibrin overlay methods for the detection of single transformed cells and colonies of transformed cells. Cell 1975; 5: 323-329
    CrossrefPubMedGoogle Scholar
  • 24 Unkeless JC, Tobia A, Ossowski L, Quigley JP, Rifkin DB, Reich E. An enzymatic function associated with transformation of fibroblasts by oncogenic viruses I. Chick embryo fibroblast cultures transformed by avian RNA tumor viruses. J Exp Med 1973; 137: 85-111
    CrossrefPubMedGoogle Scholar
  • 25 Laki K. The polymerization of proteins: the action of thrombin on fibrinogen. Arch Biochem Biophys 1951; 32: 317-324
    CrossrefPubMedGoogle Scholar
  • 26 Moza AK, Kumar M, Sapru RP. Satisfactory radioiodination of rabbit fibrinogen using chloramine-T. J Lab Clin Med 1976; 87: 169-178
    PubMedGoogle Scholar
  • 27 Haverkate F, Timan G. Preparation of highly purified bovine fibrin plates. In: Progress in Chemical Fibrinolysis and Thrombolysis 1976. Davidson JF, Samana MM, Desnoyers PC. (Eds) 2 67-71 Raven Press; New York: 1976
    Google Scholar
  • 28 Deutsch DG, Mertz ET. Plasminogen: purification from human plasma by affinity chromatography. Science 1970; 170: 1095-1096
    CrossrefPubMedGoogle Scholar
  • 29 Philo RD, Gaffney PJ. Assay methodology for urokinase: its use in assessing the composition of mixtures of high and low-molecular weight urokinase. Thromb Res 1981; 21: 81-88
    CrossrefPubMedGoogle Scholar
  • 30 Granelli-Pipemo A, Reich E. A study of protease-inhibitor complexes in biological fluids. J Exp Med 1978; 148: 223-234
    CrossrefPubMedGoogle Scholar
  • 31 Brakman P, Albrechtsen OK, Astrup T. A comparative study of coagulation and fibrinolysis in blood from normal men and women. Br J Haematol 1966; 12: 74-85
    CrossrefPubMedGoogle Scholar
  • 32 Knox AM, Sturton RG, Cooling J, Brindley DN. Control of hepatic triacylglycerol synthesis. Diurnal variations in hepatic phosphatidate phosphohydrolase activity and in the concentrations of circulating insulin and corticosterone in rats. Biochem J 1979; 180: 441-443
    CrossrefPubMedGoogle Scholar
  • 33 Thorsen S. Differences in the binding to fibrin of native plasminogen and plasminogen modified by proteolytic degradation. Influence of omega-amino-carboxylic acids. Biochim Biophys Acta 1975; 393: 55-65
    CrossrefPubMedGoogle Scholar
  • 34 Sodetz JM, Brockway WJ, Castellino FJ. Multiplicity of rabbit plasminogen. Physical characterization. Biochemistry 1972; 11: 4451-4458
    CrossrefPubMedGoogle Scholar
  • 35 Summaria L, Arzadon L, Bernabe P, Robbins KC. Studies on the isolation of the multiple molecular forms of human plasminogen and plasmin by isoelectric focusing methods. J Biol Chem 1972; 247: 4691-4702
    PubMedGoogle Scholar
  • 36 Wallen P, Wiman B. Characterization of human plasminogen II. Separation and partial characterization of different molecular forms of human plasminogen. Biochim Biophys Acta 1972; 257: 122-134
    CrossrefPubMedGoogle Scholar
  • 37 Allen RA, Pepper DS. Isolation and properties of human vascular plasminogen activator. Thromb Haemostas 1981; 45: 43-50
    PubMedGoogle Scholar
  • 38 Wiman B, Collen D. Molecular mechanism of physiological fibrinolysis. Nature 1978; 272: 549-550
    CrossrefPubMedGoogle Scholar
  • 39 Gurewich V, Hyde E, Lipinski B. The resistance of fibrinogen and soluble fibrin monomer in blood to degradation by a potent plasminogen activator derived from cadaver limbs. Blood 1975; 46: 555-565
    PubMedGoogle Scholar
  • 40 Mattsson C, Nyberg-Arrhenius V, Wallen P. Dissolution of thrombi by tissue plasminogen activator, urokinase and streptokinase in an artificial circulating system. Thromb Res 1981; 21: 535-545
    CrossrefPubMedGoogle Scholar
  • 41 Matsuo O, Rijken DC, Collen D. Comparison of the relative fibrinogenolytic, fibrinolytic and thrombolytic properties of tissue plasminogen activator and urokinase in vitro. Thromb Haemostas 1981; 45: 225-229
    PubMedGoogle Scholar
  • 42 Thorsen S, Glas-Greenwalt P, Astrup T. Differences in the binding to fibrin of urokinase and tissue plasminogen activator. Thrombos Diathes Haemorrh 1972; 28: 65-74
    PubMedGoogle Scholar
  • 43 Mattii R, Ambrus JL, Sokal JE, Mink I. Production of members of the blood coagulation and fibrinolysin systems by the isolated perfused liver. Proc Soc Exp Biol Med 1964; 116: 69-72
    CrossrefPubMedGoogle Scholar
  • 44 Losito R, Gattiker H, Lemieux E, Longpre B. Comparative effects of an erythrocyte enriched synthetic replenisher and whole blood on streptokinase activity in the isolated perfused liver. Thromb Res 1979; 14: 125-130
    CrossrefPubMedGoogle Scholar