Cuscutae semen has been shown to have beneficial effects in the treatment of vitiligo, recorded in the Chinese Pharmacopoeia, whereas the effects of its constituent compounds remains to be elucidated. Using a tetrazolium bromide assay, the present study found that hyperoside (0.5‑200 µg/ml) significantly increased the viability of human melanocytes in a time‑ and dose‑dependent manner. The present study used a cell model of hydrogen peroxide (H2O2)‑induced oxidative damage to examine the effect of hyperoside on human primary melanocytes. The results demonstrated that hyperoside pretreatment for 2 h decreased cell apoptosis from 54.03±9.11 to 17.46±3.10% in the H2O2‑injured melanocytes. The levels of oxidative stress in the mitochondrial membrane potential of the melanocytes increased following hyperoside pretreatment. The mRNA and protein levels of B‑cell lymphoma‑2/Bcl‑2‑associated X protein and caspase 3 were regulated by hyperoside, and phosphoinositide 3‑kinase/AKT and mitogen‑activated protein kinase signaling were also mediated by hyperoside. In conclusion, the results of the present study demonstrated that hyperoside protected the human primary melanocytes against oxidative damage.