Lu Rong, velvet antler (VA), is a traditional Chinese medicine, which is used as a food supplement and therapeutic drug in China, Japan, Russia, New Zealand and Southeast Asia. The regenerative characteristics of VA have resulted in great research interest, particularly regarding the fields of organ grafting and stem cell differentiation. Various VA proteomic studies verified that proteins act as the primary bioactive components of VA. The present study aimed to investigate if VA proteins (VA-pro) influence endothelial progenitor cell (EPC) viability. Various methods have previously been used to investigate VA-pro, including freeze-drying technology, ultrasonic wave methods, high performance liquid chromatography-mass spectrometry, EPCs extraction and culture. Results demonstrated that VA-pro promoted EPCs proliferation and migration, particularly at a concentration of 1 mg/ml. Furthermore, VA-pro increased the activation level of Notch1 intracellular domain and Hes1, and the level of phosphorylated-Akt and phosphorylated-mechanistic target of rapamycin. VA-pro may therefore affect EPC viability via regulation of the Notch and Akt signaling pathways. The present study revealed the effects and potential molecular mechanism of VA-pro on EPCs, and suggested an association between VA regeneration characteristics and the optimization of EPC viability. These findings may contribute to EPC transplantation research and aid in providing a novel treatment method for vascular diseases in the future.