In view of important implication of resveratrol and thrombin in the process of platelet aggregation and apoptosis, the interaction effect between them and its biological implication was studied. Resveratrol could decrease fluorescence emission intensities of thrombin by a dynamic collision quenching process. The binding process was spontaneous and the resveratrol-thrombin complex formation was an endothermal reaction induced by hydrophobic interaction and hydrogen. Non-radiation energy transfer occurred from thrombin to resveratrol at the molecular level. Kinetic studies showed that the fluorescence intensity of thrombin linearly decreased along with prolonged time, whereas resveratrol intensity reduced continuously with degressive plateau in the form of second exponential decay function. Moreover, the interaction caused a decrease of α-helical structure resulting in thrombin conformation alteration. In addition, the isomerization of resveratrol was not prevented but its stability was improved with the addition of thrombin. This work might give deeper insight into resveratrol as a kind of nutritional factor with the inhibition of platelet aggregation and stimulation of platelet apoptosis in the functional food.