Fig. 1

Oroxylin A inhibits thrombus formation. A, OroA inhibited thrombus formation in mesenteric arterioles of wild-type mice. Wild-type mice randomly received 0.1% DMSO (once per day, o.g.) or OroA (20 or 40 mg/kg, once per day, o.g.), or aspirin (20 mg/kg, once per day, o.g.) for 3 days before the experiment. FeCl₃-induced thrombosis was performed and the thrombus area was recorded. Representative images at different time points are shown. Statistical analysis of thrombosis assessed using area of thrombus at indicated times and time to first thrombus (> 20 μm) are shown (n = 8). Scale bar = 100 μm. B, OroA inhibited thrombus formation over an immobilized collagen surface at a shear rate of 1000 s^− 1 in whole blood from wild-type mice. After treatment with 0.1% DMSO (once per day, o.g.), OroA (20 or 40 mg/kg, once per day, o.g.), or aspirin (20 mg/kg, once per day, o.g.), whole blood was perfused through collagen-coated bioflux plates. Representative images of thrombus formation at the indicated time points are presented (n = 6). Scale bar = 100 μm. C, OroA reduced the number of pulmonary microthrombus and platelet infiltration in mice with pulmonary embolism. Wild-type mice randomly received 0.1% DMSO (once per day, o.g.) or OroA (40 mg/kg, once per day, o.g.) for 3 days before the experiment. The mixture of collagen (100 µg/kg) and epinephrine (600 µg/kg) was used to induce pulmonary embolism. After 5 h, the mice were anesthetized and the lungs were taken for H&E and CD61 (platelet marker) staining. Representative images and quantification of number of thrombi observed in a random 10× field or CD61 positive areas expressed as a percentage of the field, respectively (n = 6). Col, collagen; Epi, epinephrine. Scale bar = 100 μm (high magnification view) and 2 mm (low magnification view). D, OroA slightly prolonged the tail bleeding time in mice. Wild-type mice randomly received 0.1% DMSO (once per day, o.g.), OroA (40 mg/kg once per day, o.g.), or aspirin (20 mg/kg once per day, o.g.) for 3 days before the experiment. Bleeding time and tail blood loss were measured, and each dot represents a single mouse (n = 12).Statistical analyses were performed using one-way ANOVA followed by Tukey’s multiple comparison test in (A and D), and two-way ANOVA followed by Sidak’s multiple comparison test in (B), and unpaired two-tailed Student’s t-test in (C). NS, no significance; *P < 0.05; **P < 0.01; ***P < 0.001