It has been shown that oxidative damage and inflammation caused by hyperglycemia in endothelial cells are key factors triggering diabetic vascular complications. The aim of the present study was to investigate the antioxidant and anti‑inflammatory effects of Danhong Huayu Koufuye (DHK)‑medicated serum on high glucose (HG)‑induced injury in endothelial cells, and examine its underlying mechanisms. EA. hy926 cells were treated with normal glucose, HG, or HG with DHK‑medicated serum. Cell viability was assessed using the MTT method. Apoptosis was detected using flow cytometry. Intracellular reactive oxygen species (ROS) levels were measured using the 2',7'‑dichlorodihydrofluorescein method. Cell culture supernatant was collected for detecting the activities of glutathione peroxidase (GPx) and superoxide dismutase (SOD), and the levels of malondialdehyde (MDA). The protein expression levels of intercellular adhesion molecule‑1 (ICAM‑1), nuclear factor‑κB (NF‑κB), hypoxia‑inducible factor‑1α (HIF‑1α) and vascular endothelial growth factor (VEGF) were determined using western blot analysis. The results revealed that DHK‑medicated serum accelerated the proliferation and inhibited the apoptosis of cells treated with HG (P<0.01) in a dose‑dependent manner. Compared with the HG group, the high levels of ROS and MDA were significantly reduced by DHK‑medicated serum (P<0.01). A 10% concentration of DHK‑medicated serum increased the activities of SOD and GPx by 59.4 and 95.5%, respectively. The high protein expression levels of ICAM‑1, NF‑κB, VEGF and HIF‑1α were significantly ameliorated by DHK‑medicated serum (P<0.01, vs. HG group). These findings indicated that DHK‑medicated serum protected EA. hy926 cells from HG‑induced injury and apoptosis through antioxidation and anti‑inflammatory effects.