Thromb Haemost 1993; 69(04): 394-396
DOI: 10.1055/s-0038-1651619
Original Article
Platelets
Schattauer GmbH Stuttgart

Microbubble-Induced Phospholipase C Activation Does not Correlate with Platelet Aggregation

R Malmgren
1   The Department of Experimental Surgery Karolinska Institute, Stockholm, Sweden
,
T Thorsen
2   The Department of Biochemistry, University of Bergen, Norway
,
A Nordvik
2   The Department of Biochemistry, University of Bergen, Norway
,
H Holmsen
2   The Department of Biochemistry, University of Bergen, Norway
› Author Affiliations
Further Information

Publication History

Received 24 June 1992

Accepted after revision 14 December 1992

Publication Date:
05 July 2018 (online)

Summary

The effect of nitrogen-(N2-)microbubbles on platelets resembles that of common platelet agonists with respect to aggregation and secretion, but is considerably slower and is poorly inhibited by aspirin. This paper reports the effect of microbubbles on platelet phospholipase C activity in gelfiltered human platelets prelabelled with [32P]Pi ([32P]-GFP). The experiments were run in the presence of an ADP scavenging system in order to rule out effects of ADP. Stimulation of [32P]-GFP for 30 min with microbubbles caused a significant reduction in single platelets (p <0.0004) and a significant increase in 32P-activity in the phosphatidic acid (PA) fraction (p <0.02). Epinephrine potentiated the microbubble-induced reduction in single platelets (p <0.05), but did not enhance the amount of 32P in the platelet [32P]PA fraction. The 32P-radioactivity in the PI-fraction increased with time to a similar extent when [32P]-GFP was stirred for 30 min in absence of microbubbles as it did after 30 min of agonist exposure. There were no significant changes in the [32P]PIP and [32P]PIP2 fractions. Aspirin abolished the microbubble-induced increase in 32P-activity in the PA fraction, but had no significant effect on the reduction in single platelets. Aspirin had a small but significant, reducing effect on platelet aggregation induced by a combination of epinephrine and microbubbles (p <0.05). With epinephrine, however, aspirin did not completely abolish the increase in [32P]-PA. It is concluded that microbubbles alone cause platelets to aggregate by a novel mechanism that operates independent of cyclooxygenase-dependent arachidonic acid metabolites and phospholipase C activation.

 
  • References

  • 1 Cockett ATK, Pauley SM, Zehl DN, Pilmanis AA, Cockett WS. Pathophysiology of bends and decompression sickness. Arch Surg 1979; 114: 296-301
  • 2 Philp RB, Schacham P, Gowdey CW. Involvement of platelets and microthrombi in experimental decompression sickness: Similarities with disseminated intravascular coagulation. Aerospace Med 1971; 42: 494-502
  • 3 Strauss RH. Diving medicine. Am Rev Respir Dis 1979; 119: 1001-1023
  • 4 Osada H, Ward CA, Duffin J, Nelems JM, Cooper JD. Microbubble elimination during priming improves biocompability of membrane oxygenators. Am J Physiol 1978; 234: 646-652
  • 5 Ward CA, Ruegsegger B, Stanga D, Zingg W, Herbert MA. Relation of proteins, platelets, and gas nuclei in adhesion to a synthetic material. Am J Physiol 1977; 233: 100-105
  • 6 Thorsen T, Brubakk A, Ovstedal T, Farstad M, Holmsen H. A method for production of N2-microbubbles in platelet rich plasma in an aggregometer-like apparatus, and effect on the platelet density in vitro. Undersea Biomed Res 1986; a 13: 271-288
  • 7 Thorsen T, Dalen H, Bjerkvig R, Holmsen H. Transmission and scanning electron microscopy of N2 microbubble-activated human platelets in vitro. Undersea Biomed Res 1987; 14: 45-59
  • 8 Malmgren R, Thorsen T, Lie RT, Holmsen H. Microbubble-induced serotonin secretion in human platelets. Thromb Haemostas 1991; 65: 399-402
  • 9 Thorsen T, Lie RT, Holmsen H. Induction of platelet aggregation in vitro by microbubbles of nitrogen. Undersea Biomed Res 1989; 16: 453-463
  • 10 Thorsen T, Övstedal A, Vereide A, Holmsen H. Effects of platelet antagonists on the reduction in platelet density caused by microbubbles in vitro. Undersea Biomed Res 1986; b 13: 289-302
  • 11 Thorsen T. Gas bubbles. A “new” platelet agonist. Dissertation. University of Bergen; Bergen: 1989
  • 12 Tysnes O-B, Verhoeven AJM, Holmsen H. Phosphate turnover of phosphatidylinositol in resting and thrombin stimulated platelets. Biochim Biophys Acta 1986; 889: 183-191
  • 13 Tysnes O-B, Verhoeven AJM, Holmsen H. Rates of production and consumption of phosphatidic acid upon thrombin stimulation of human platelets. Eur J Biochem 1988; 174: 75-79
  • 14 Siess W. Molecular mechanisms of platelet activation. Physiol Rev 1989; 69: 58-178
  • 15 Steen VM, Tysnes OB, Holmsen H. Synergism between thrombin and adrenaline (epinephrine) in human platelets. Marked potentiation of inositol phospholipid metabolism. Biochem J 1988; 253: 581-586
  • 16 Malmgren R, Thorsen T, Holmsen H. Secretion from platelet dense bodies and alpha granules exhibit differences in cyclooxygenase dependence. To be published