Objective To develop a microfluidic chip analysis system which can be used to evaluate the inhibitory effect of antiplatelet drugs on platelet activation and aggregation induced by pathological high shear. Methods Polydimethoxane (PDMS)-glass microchannel chip was fabricated by soft lithography. The anticoagulant human whole blood was flowed through the microchannel chip at 1 500 s-1 shear rate, and the platelet aggregation behavior downstream of the narrow channel was photographed by fluorescence inversion microscope. The platelet aggregation coverage rate, total aggregation area, average size of thrombus and number of thrombus were obtained by image analysis. The whole blood flowing through the microfluidic chip was collected and the expression of platelet surface activation markers (P-selectin and PAC-1) was analyzed by flow cytometry. The inhibitory effect of antiplatelet drugs (aspirin, tirofiban, tirofiban) on platelet aggregation and activation downstream of stenosis was also evaluated. Results With the increase of input shear rate, the ability of platelet aggregation increased gradually. Aspirin could not inhibit platelet activation and aggregation induced by pathological high shear, while tigrel and tirofiban could significantly inhibit platelet activation and aggregation. Conclusions The microfluidic chip model developed in this study can be used to simulate the local characteristics of arterial stenosis and study the effects of different shear force gradients on platelet aggregation, as well as to evaluate the inhibitory effect of antiplatelet drugs on platelet activation and aggregationunder pathological high shear stress. Different from the traditional turbidimetry and thromboelastography in the detection of platelet function under static conditions, this study provides an in vitro model closer to the real flow environment in patients with arterial thrombotic diseases, as well as a related analysis method for the detection of platelet function in patients with percutaneous coronary intervention (PCI) or atherosclerosis.