Objective To establish a zebrafish model of thrombosis induced by three kinds of inducers and observe the anti-thrombotic effect of a Chinese traditional medicine, Guanxinning tablet (GXN). Methods The zebrafish models of thrombosis was induced by using 1.5 μmol/L phenyl hydrazine, 80 μmol/L arachidonic acid and 5 mg/L ponatinib, respectively, and were treated with various concentration of GXN, clopidogrel or asprin. The thrombus in the tail vein was observed under microscope, Erythrocytes in the zebrafish heart were stained with o-dianisidine and the erythrocyte staining intensity was assessed with a NIS-Elements DTM image analyzer, and the anti-thrombolic effect of GXN was calculated.Results Venous thrombus was significantly increased and the staining intensities of erythrocytes in the heart were significantly decreased after induction by phenyl hydrazine, arachidonic acid or Ponatinib (P<0.001), respectively. At the same time, GXN showed an incresing anti-thrombolic effect in the zebrafish models (P<0.001) in a dose-effect manner, with a IC₅₀ of GXN of 44.32 mg/L,138.5 mg/L and 459.5 mg/L, respectively. Conclusions The zebrafish models of thrombosis are successfully established by phenyl hydrazine, arachidonic acid or Ponatinib, respectively, by different formation mechanisms. GXN has been shown to have an anti-thrombosis effect, probably, by multiple target effects.