Blockade by ruthenium red of tissue factor-initiated coagulation

1 The ability of ruthenium red (RuR) to inhibit tissue factor (TF)-initiate d blood coagulation was demonstrated at the protein and cellular levels as well as in human plasma. 2 In a single-stage clotting assay, RuR concentration-dependently inhibited rabbit brain thromboplastin (rbTF)-induced coagulation and offset bacteria l endotoxin (LPS)-induced monocytic TF (mTF) hypercoagulation; the IC(50)s were estimated at 7.5 and 12.3 muM, respectively. 3 A 15-min preincubation of RuR with rbTF or monocyte suspension resulted i n the pronounced inhibition with a significantly lowered IC50 at 1.8 or 7.7 muM for rbTF or mTF procoagulation, respectively. The differences in IC(50 )s between rbTF and mTF without or with the preincubation indicated that TF was a primary target for RuR action. 4 The effect of RuR on the physiological function of TF in FVII activation was demonstrated by the proteolytic cleavage of FVII zymogen to its active forms of serine protease on Western blotting analyses. RuR readily blocked TF-catalyzed FVII activation (diminished FVIIa formation), thus down regula ting the initiation of blood coagulation. 5 Inclusion of RuR into human plasma samples in vitro significantly prolong ed prothrombin time, indicating the depressed coagulation. FVII activity wa s inhibited by 30-60% depending on the dose; as a result, FX activity also decreased. However, RuR showed no effect on thrombin time. Thus, RuR inhibi ted FVII activation to block the initiation of coagulation.