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Thromb Haemost 1989; 62(03): 1034-1039
DOI: 10.1055/s-0038-1651048
Original Article
Schattauer GmbH Stuttgart

Prostanoid Release from Ex Vivo Perfused Canine Arteries and Veins: Effects of Prolonged Perfusion, Intermittent Perfusion, as well as Exposure to Exogenous Arachidonic Acid, Thrombin and Bradykinin

Jan S Brunkwall
*   The Department of Surgery, University of Lund, Malmö General Hospital, Malmö, Sweden
,
James C Stanley
**   The Section of Vascular Surgery, Department of Surgery, University of Michigan Medical School, Ann Arbor, Michigan, USA
,
Timothy F Kresowik
**   The Section of Vascular Surgery, Department of Surgery, University of Michigan Medical School, Ann Arbor, Michigan, USA
,
Linda M Graham
**   The Section of Vascular Surgery, Department of Surgery, University of Michigan Medical School, Ann Arbor, Michigan, USA
,
William E Burkel
***   The Department of Anatomy and Cell Biology, University of Michigan Medical School, Ann Arbor, Michigan, USA
,
David Bergqvist
*   The Department of Surgery, University of Lund, Malmö General Hospital, Malmö, Sweden
› Author Affiliations
Further Information

Publication History

Received 21 December 1988

Accepted after revision 11 July 1989

Publication Date:
30 June 2018 (online)

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Summary

Regulation of prostanoid release from ex vivo perfused vessel segments is not fully understood. A series of perfusion experiments were performed with canine arteries and veins to define certain regulatory phenomena. Arteries were perfused with pulsatile flow of 90 ml/min at a pressure of 100 mmHg, and veins with nonpulsatile flow of 90 ml/min at a pressure of 7 mmHg. Segments were perfused with Hanks' balanced salt solution for five 15-min periods with the perfusate exchanged after each study period. With onset of perfusion, there was an initial burst of prostacyclin release to 127 ± 40 pg/mm2, declining to 32 ± 10 pg/mm2 after 60 minutes (p <0.005). If perfusion continued for 5.5 hours, there was a stable release period between 1 and 3 hours, followed by a very slow decline. At that time addition of arachidonic acid (AA) increased prostacyclin release six-fold (p <0.01). Vessels perfused for 1 hour and then rested for another hour, responded to reperfusion at the second onset of flow with a two-fold increase in prostacyclin release (p <0.01). Vessels perfused with thrombin, bradykinin or A A (either added to each perfusate or only to the last perfusate) exhibited greater prostacyclin release than did control segments. Release of thromboxane steadily declined with time in all parts of the study, and only increased with the addition of A A to the perfusate. These data indicate that vessel segments subjected to ex vivo perfusion do not maximally utilize enzyme systems responsible for prostanoid production, and after 1 hour perfusion have not depleted their phospholipids, and maintain functioning levels of phospholipase and cyclooxygenase activity. This perfusion model allows for the study of prostacyclin and thromboxane release from arteries and veins and their response to various drugs and other stimuli.

Keywords

Vessel wall - Prostanoid release - Arachidonic acid - Thrombin - Bradykinin
 

  • References

  • 1 Hajjar DP, Weksler BB, Falcone D, Hefton JM, Tack-Goldman K, Minick CR. Prostacyclin modulates cholesteryl ester hydrolytic activity by its effect on cyclic adenosine monophosphate in rabbit aortic smooth muscle cells. J Clin Invest 1982; 70: 479-488
    CrossrefPubMedGoogle Scholar
  • 2 Hajjar DP, Weksler BB. Metabolic activity of cholesteryl esters in aortic smooth muscle cells is altered by prostaglandins I2 and E2 . J Lipid Res 1983; 24: 1176-1185
    PubMedGoogle Scholar
  • 3 MacIntyre DE, Pearson JD, Gordon JL. Localization and stimulation of prostacyclin production in vascular cells. Nature 1978; 271: 549-551
    CrossrefPubMedGoogle Scholar
  • 4 Moncada S, Herman AG, Higgs EA, Vane JR. Differential formation of prostacyclin (PGX or PGI2) by layers of the arterial wall. An explanation for the anti-thrombotic properties of vascular endotheliumboxane as well as prostacyclin. Am J Physiol 1983; 244: R839-R843
    PubMedGoogle Scholar
  • 5 Sinzinger H, Firbas W. Gefäßwandzellen synthetisieren Thromboxan. Wiener Klin Wschr 1983; 95: 758-760
    PubMedGoogle Scholar
  • 6 Ingerman-Wojenski C, Silver MJ, Smith JB, Macarack E. Bovine endothelial cells in culture produce thromboxane as well as prostacyclin. J Clin Invest 1981; 67: 1292-1296
    CrossrefPubMedGoogle Scholar
  • 7 Weksler BB, Ley CW, Jaffe EA. Stimulation of endothelial prostacyclin production by thrombin, trypsin, and the ionophore A 23187. J Clin Invest 1978; 62: 923-930
    CrossrefPubMedGoogle Scholar
  • 8 Egan RW, Paxton J, Kuehl Jr FA. Mechanism for irreversible deactivation of prostaglandin synthetase. J Biol Chem 1976; 251: 7329-7335
    PubMedGoogle Scholar
  • 9 Hemler ME, Lands WE M. Evidence for a peroxide-initiated free radical mechanism of prostaglandin biosynthesis. J Biol Chem 1980; 255: 6253-6261
    PubMedGoogle Scholar
  • 10 Dejana E, Balconi G, DeCastellamau C, Barbieri B, Vergara-Dauden M, DeGaetano G. Prostacyclin production by human endothelial and bovine smooth muscle cells in culture. Effect of repeated stimulation with arachidonic acid, thrombin and ionophore A 23187. Biochim Biophys Acta 1983; 750: 261-267
    CrossrefPubMedGoogle Scholar
  • 11 Bjøro K, Hovig T, Stokke RT, Stray-Pedersen S. Formation of prostanoids in human umbilical vessels perfused in vitro. Prostaglandins 1986; 31: 683-698
    CrossrefPubMedGoogle Scholar
  • 12 Papp AC, Crowe L, Pettigrew LC, Wu KK. Production of eicosanoids by deendothelialized rabbit aorta: Interaction between platelets and vascular wall in the synthesis of prostacyclin. Thromb Res 1986; 42: 549-556
    CrossrefPubMedGoogle Scholar
  • 13 Kristensen SD, Amfred T, Dyerberg J. Eicosapentaenoic acid potentiates the production of prostacyclin-like material in the arachidonic acid perfused human umbilical vein. Thromb Res 1984; 36: 305-314
    CrossrefPubMedGoogle Scholar
  • 14 Kent RS, Diedrich SL, Whorton AR. Regulation of vascular prostaglandin synthesis by metabolites of arachidonic acid in perfused rabbit aorta. J Clin Invest 1983; 72: 455-465
    CrossrefPubMedGoogle Scholar
  • 15 Voss BV, ten Hoor F, Haddeman E, Don JA. Influence of dietary linoleic acid on the prostacyclin production of the isolated pulsatingly perfused rabbit aorta. Prostaglandins Leukotrienes Med 1982; 8: 503-516
    PubMedGoogle Scholar
  • 16 Brunkwall JS, Stanley JC, Graham LM, Burkel WE, Bergqvist D. Comparison of static incubation versus physiologic perfusion techniques for quantitation of luminal release of prostacyclin and thromboxane in canine arteries and veins. J Surg Res 1988; 45: 1-7
    CrossrefPubMedGoogle Scholar
  • 17 Quadt JF A, Voss R, ten Hoor F. Prostacyclin production of the isolated pulsatingly perfused rat aorta. J Pharmacol Methods 1982; 7: 263-270
    CrossrefPubMedGoogle Scholar
  • 18 Brunkwall JS, Stanley JC, Graham LM, Burkel WE. Influence of pressure, flow rate, and pulsatility on release of 6-keto-PGF1α and thromboxane B2 in ex vivo-perfused canine veins. J Vase Surg 1988; 7: 99-107
    PubMedGoogle Scholar
  • 19 Brunkwall JS, Stanley JC, Graham LM, Burkel WE, Bergqvist D. Arterial 6-keto-PGF1α and TxB2 release in ex vivo perfused canine vessels: Effects of pulse rate, pulsatility, altered pressure and flow rate. Eur J Vasc Surg 1989; 3: 219-225
    CrossrefPubMedGoogle Scholar
  • 20 Nation JL. A new method using hexamethyldisilazane for preparation of soft insect tissues for scanning electron microscopy. Stain Technol 1983; 58: 347-351
    PubMedGoogle Scholar
  • 21 Simmons PM, Salmon JA, Moncada S. The release of leukotriene B4 during experimental inflammation. Biochem Pharmacol 1983; 32: 1353-1359
    CrossrefPubMedGoogle Scholar
  • 22 Fitzpatrick FA, Gorman RR, McGuire JC, Kelly RC, Wynalda MA, Sun FF. A radioimmunoassay for thromboxane B2 . Analyt Biochem 1977; 82: 1-7
    CrossrefPubMedGoogle Scholar
  • 23 Brox JH, Nordoy A. Prostacyclin and 51Cr release in cultured endothelial cells. Hemostasis 1982; 12: 345-352
    PubMedGoogle Scholar
  • 24 Mehta P, Mehta J, Hay D. Thromboxane and prostacyclin generation by intact human vessels in response to balloon catheter trauma. Prostaglandins Leukotrienes Med 1982; 9: 539-548
    CrossrefPubMedGoogle Scholar
  • 25 Boeynaems JM, Galand N, Ketelbant P. Prostacyclin production by the de-endothelialized rabbit aorta. J Clin Invest 1985; 76: 7-14
    CrossrefPubMedGoogle Scholar
  • 26 Goldsmith JC. Contribution of the subendothelium to prostacyclin release after vascular injury. J Lab Clin Med 1982; 100: 574-584
    PubMedGoogle Scholar
  • 27 Brotherton AF A, Hoak JC. Prostacyclin biosynthesis in cultured vascular endothelium is limited by deactivation of cyclooxygenase. J Clin Invest 1983; 72: 1255-1261
    CrossrefPubMedGoogle Scholar
  • 28 Lollar P, Owen WG. Evidence that the effects of thrombin on arachidonate metabolism in culture human endothelial cells are not mediated by a high affinity receptor. J Biol Chem 1980; 255: 8031-8034
    PubMedGoogle Scholar
  • 29 Ragab-Thomas JM F, Hullin F, Chap H, Douste-Blazy L. Pathways of arachidonic acid liberation in thrombin and calcium ionophore A 23187-stimulated human endothelial cells: respective roles of phospholipids and triacylglycerol and evidence for diacylglycerol generation from phosphatidylcholine. Biochim Biophys Acta 1987; 917: 388-397
    CrossrefPubMedGoogle Scholar
  • 30 Long SL, McLaughlin NJ, Tzeng Cy, Patton G. Prostacyclin synthesis and deacylation of phospholipids in human endothelial cells: comparison of thrombin, histamin and ionophore A 23187. Thromb Res 1985; 38: 1-10
    CrossrefPubMedGoogle Scholar
  • 31 Goldsmith JC, Jafvert CT, Lollar P, Owen WG, Hoak JC. Prostacyclin release from cultured and ex vivo bovine vascular endothelium. Studies with thrombin, arachidonic acid, and ionophore A 23187. Lab Invest 1981; 45: 191-197
    PubMedGoogle Scholar
  • 32 Goldsmith JC, Kisker CT. Thrombin-endothelial interactions: Critical importance of endothelial cell vessel of origin. Thromb Res 1982; 25: 131-136
    CrossrefPubMedGoogle Scholar
  • 33 Malik AB, Lo SK, Bizios R. Thrombin induced alterations in endothelial permeability. Ann NY Acad Sci 1986; 485: 293-309
    CrossrefPubMedGoogle Scholar
  • 34 Killackey JJ F, Johnston MG, Movat HZ. Increased permeability of microcarrier-cultured endothelial monolayers in response to histamine and thrombin. A model for the in vitro study of increased vasoperme-ability. Am J Pathol 1986; 122: 50-56
    PubMedGoogle Scholar
  • 35 De Groot PG, Brinkman HJ M, Gonsalves MD, van Mourik JA. The role of thrombin in the regulation of endothelial prostaglandin production. Biochim Biophys Acta 1985; 846: 342-349
    CrossrefPubMedGoogle Scholar