Effects of simvastatin on the inhibition of (A and B) platelet aggregation, (C) FITC-triflavin binding to the α
integrin and (D) [Ca2+]i mobilization in activated platelets. Washed platelets (3.6 × 108/ml) were preincubated with simvastatin (10-100 μM) or 0.5% DMSO, followed by the addition of collagen (1 μg/ml, open circle), U46619 (1 μM, upside down open triangle), thrombin (0.02 U/ml, open square) or arachidonic acid (60 μM, open diamond) to trigger platelet aggregation (A and B) and the ATP-release reaction (A, upper tracings) or (D) [Ca2+]i mobilization. (C) The solid line represents the fluorescence profiles of FITC-triflavin (2 μg/ml) (a) with or (b) without EDTA (5 mM); or pretreatment of simvastatin (c) (30 μM) and (d) (50 μM), followed by the addition of FITC-triflavin (2 μg/ml). Data are presented as the means ± S.E.M. (n = 4); ***P < 0.001, compared to the control group; #P < 0.05 and ##P < 0.01, compared to the collagen group.