Research Article

Antiplatelet and Antithrombotic Effects of Epimedium koreanum Nakai

Figure 3

E. koreanum inhibits integrin αIIbβ3-mediated inside-out and outside-in signaling. ((a) and (b)) Flow cytometric measurements of fibrinogen binding in rat platelets (3 × 108 cells/mL) treated with vehicle or different concentrations of E. koreanum fraction (EAF), or EGTA ((A) resting, (B) vehicle, (C) 50 μg/mL, (D) 100 μg/mL, (E) 200 μg/mL, and (F) 10 μM EGTA)), and stimulated with collagen (b–f). (c) Results of fibronectin adhesion assay, which was performed using an assay kit according to the manufacturer’s instructions and by following the procedure described in the methods section. (d) In vitro effect of E. koreanum on clot retraction for 2 h at room temperature after thrombin addition and photographed at 15 min intervals. Representative images of clot retraction at 90 min after thrombin addition with or without EAF. Y-27632 (ROCK inhibitor) was used as a control. Lanes 1–5 correspond to lanes 1–5 in (e). (f) Clot retraction kinetics were measured by ImageJ software, and the clot surface areas were plotted as a percentage of retraction. Bar graphs summarizing the inhibitory effect of EAF on fibrinogen binding to integrin αIIbβ3 (B), fibronectin adhesion (C), clot retraction (E), and kinetics of clot retraction (F). Results are shown as mean ± SD values from at least four independent experiments. , and versus control. R: resting; C: control; GR: GR155053; Y-2: Y-27632.
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