Combination of Colchicine and Ticagrelor Inhibits Carrageenan-Induced Thrombi in Mice

Figure 3

Colchicine inhibits platelet activation in vivo and thrombin-induced platelet activation in vitro. (a, b) Mice received i.g. administration of colchicine (0.05 mg/day/kg bodyweight) or PBS for 2 days, followed by collection of blood samples. Platelets were collected from mouse blood, then detected expression of CD41 (a) and CD36 (b) by immunofluorescent staining; (c, d) human platelets were treated with CaCl₂ (1 mM, control group) or CaCl₂ plus colchicine (100 ng/mL, Col group) at 37°C for 30 min, then added fibrinogen (2 mg/mL) with thoroughly mixed. Thrombin (1 U/mL) was added to the mixture to trigger clot retraction, and clots were photographed at the different time points (c). The spreading area of platelet clot was measured using Photoshop CS6 software (d); (e–h) human platelet suspension was pretreated with colchicine (100 ng/mL), LY294002 (10 μM), U0126 (5 μM), colchicine plus LY294002, or colchicine plus U0126 for 2 h at 37°C, followed by treatment with thrombin (1 U/mL) for 2 min. Expression of p-AKT, AKT, p-ERK1/2, ERK1/2, and GAPDH was determined by Western blot (e, g) with quantitative analysis of band density (f, h). , , and vs. control; ^#, ^##, and ^### vs. thrombin-treated group ().

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