Estriol-induced fibrinolysis due to the activation of plasminogen to plasmin by nitric oxide synthesis in platelets.
Estriol, an oestrogen, at 0.6 nmol/l was reported to inhibit ADP-induced platelet aggregation through nitric oxide synthesis. As nitric oxide has been reported to cause fibrinolysis due to the activation of plasminogen to plasmin, the role of estriol as a fibrinolytic agent was investigated. Also, the mechanism of estriol-induced nitric oxide synthesis in anucleated platelets was investigated. The estriol-induced lysis of platelet-rich plasma (PRP) clot was determined by photography of the clot lysis and by the assay of fibrin degradation products in the lysate and was obtained by SDS-PAGE. Nitric oxide was determined by methemoglobin method. The platelet membrane protein was isolated from the platelets by using Triton X-100 (0.05% v/v). The binding of estriol to the protein was determined by Scatchard plot by using an ELISA for estriol. Estriol at 0.6 nmol/l was found to lyse the clotted PRP due to fibrinolysis that produced fibrin degradation products in the lysate. The amino acid analysis of the platelet membrane protein, which resembles with nitric oxide synthase (NOS) activity, was activated nearly 10-fold over the control in the presence of estriol and was identified to be a human serum albumin precursor (Mr. 69 kDa) that binds to estriol with Kd1 of 6.0 × 10 mol/l and 39 ± 2 molecules of estriol bound the NOS molecule. The estriol-induced nitric oxide is capable of inducing fibrinolysis of the clotted PRP. The binding of estriol to platelet membrane NOS activated the enzyme in the absence of DNA in the platelet.
aSinha Institute of Medical Science and Technology, Kolkata bPost Graduate Department of Biochemistry and Biotechnology, Cell and Molecular Therapeutics Laboratory, Oriental Institute of Science and Technology, Vidyasagar University, Medinipur, West Bengal, India cJames J. Peters VA Medical Center, Bronx, New York, USA.
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D000328:Adult; D001792:Blood Platelets / Q000378:metabolism*; D004789:Enzyme Activation / Q000187:drug effects; D004964:Estriol / Q000378:metabolism / Q000494:pharmacology*; D005260:Female; D005338:Fibrin Fibrinogen Degradation Products / Q000032:analysis; D005341:Fibrinolysin / Q000378:metabolism*; D005342:Fibrinolysis / Q000187:drug effects*; D006801:Humans; D008297:Male; D008565:Membrane Proteins / Q000097:blood* / Q000302:isolation & purification; D009569:Nitric Oxide / Q000096:biosynthesis* / Q000097:blood; D010958:Plasminogen / Q000096:biosynthesis*; D010974:Platelet Aggregation; D053657:Platelet-Rich Plasma; D011485:Protein Binding; D011498:Protein Precursors / Q000378:metabolism; D012709:Serum Albumin / Q000378:metabolism; D055815:Young Adult
D005338:Fibrin Fibrinogen Degradation Products; D008565:Membrane Proteins; D011498:Protein Precursors; D012709:Serum Albumin; D009569:Nitric Oxide; D010958:Plasminogen; D005341:Fibrinolysin; D004964:Estriol