A Naphthalenic Derivative ND-1 Inhibits Thrombus Formation by Interfering the Binding of Fibrinogen to Integrin αIIbβ3

Figure 2

Effect of ND-1 on the binding of fibrinogen to integrin αIIbβ3 and P-selectin expression. (a) Effect of ND-1 on fibrinogen/integrin αIIbβ3 ELISA. Purified human integrin αIIbβ3 with or without ND-1 was added to wells coated with fibrinogen for 2 h at 37°C, followed by adding mouse anti-human integrin β3 antibody. The binding of fibrinogen to αIIbβ3 was determined using anti-mouse IgG-conjugated alkaline phosphatase and disodium 4-nitrophenyl substrate at 405 nm. All experiments were performed in triplicate. ((b) to (e)) Effect of ND-1 on P-selectin expression on platelets. Platelet-rich plasma was preincubated with ND-1 (50 μM) or vehicle for 10 min at 37°C. Activation was initiated by the addition of 20 μM ADP (b), 1 μg/mL collagen (c), 0.25 U/ml thrombin (d), or 2 μM U46619 (e) for 10 min at 37°C. P-selectin expression on platelet surface was detected by flow cytometry.

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