Insulin-induced release of plasminogen activator from human blood platelets.
Incubation of washed platelets in Tyrode buffer, pH 7.5, with insulin (200 microU/ml) and CaCl2 (1.2 mM) at 37 degrees C for 3 h resulted in a threefold increase of plasminogen activator activity in the supernatant over the basal level as determined by both the amidolytic assay and the proteolysis of alpha-casein through the formation of plasmin from plasminogen. This plasminogen activator showed no plasmin-like activity and was inhibited by anti-tissue plasminogen activator antibody as well as by type 1 plasminogen activator inhibitor. The substrate specificity and the inhibition of the enzymic activity by various inhibitors indicated that the platelet plasminogen activator (pPA) was related to tissue-type plasminogen activator of relative molecular weight 56,000. Fibrinolytic activity of pPA and its insulin-dependent release were demonstrated by the shortening of euglobulin lysis time and by the clot lysis time of platelet-rich plasma from normal and type I diabetes mellitus patients. Treatment of platelet membranes with insulin also increased the release of pPA. Increased levels of adenosine 3',5'-cyclic monophosphate (cAMP) in platelets by incubation with various agents completely inhibited the insulin-induced release of the activator. On the other hand, inhibition of platelet aggregation by aspirin had no effect on the release of pPA, indicating that the effect of cAMP was not due to the inhibition of platelet aggregation by the nucleotide.
Kahn
N N
NN
Department of Medicine, Mount Sinai Medical Center, New York 10029.
Bauman
W A
WA
Sinha
A K
AK
D000527:Alprostadil / Q000494:pharmacology; D001241:Aspirin / Q000494:pharmacology; D001792:Blood Platelets / Q000187:drug effects / Q000201:enzymology*; D003994:Bucladesine / Q000494:pharmacology; D002122:Calcium Chloride / Q000494:pharmacology; D002462:Cell Membrane / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D066298:In Vitro Techniques; D007328:Insulin / Q000494:pharmacology*; D007700:Kinetics; D010958:Plasminogen / Q000494:pharmacology; D017395:Plasminogen Activator Inhibitor 1 / Q000494:pharmacology; D010960:Plasminogen Activators / Q000037:antagonists & inhibitors / Q000097:blood* / Q000302:isolation & purification; D010975:Platelet Aggregation Inhibitors / Q000494:pharmacology*; D011972:Receptor, Insulin / Q000378:metabolism; D011994:Recombinant Proteins / Q000494:pharmacology; D013806:Theophylline / Q000494:pharmacology; D013997:Time Factors; D000979:alpha-2-Antiplasmin / Q000494:pharmacology
D007328:Insulin; D017395:Plasminogen Activator Inhibitor 1; D010975:Platelet Aggregation Inhibitors; D011994:Recombinant Proteins; D000979:alpha-2-Antiplasmin; D003994:Bucladesine; D010958:Plasminogen; D013806:Theophylline; D011972:Receptor, Insulin; D010960:Plasminogen Activators; D000527:Alprostadil; D002122:Calcium Chloride; D001241:Aspirin