Parthenolide is a natural product from the shoots of Tanacetum parthenium that has been demonstrated to have immunomodulatory effects in a number of diseases. The present study aimed to determine the effect and mechanism of parthenolide on the apoptotic ability of H2O2‑induced osteoblasts. Cell viability was analyzed with a MTT assay and the apoptotic rate was subsequently measured using flow cytometry. The activity of the antioxidative enzymes superoxide dismutase (SOD) and glutathione peroxidase (GPX), and the serum marker enzymes alkaline phosphatase (ALP), malondialdehyde (MDA) and lactate dehydrogenase (LDH) was measured. Reverse transcription‑quantitative polymerase chain reaction and western blot analyses were performed to analyze the expression levels of osteogenesis and oxidative stress‑associated genes. The results indicated that parthenolide increased cell viability and inhibited the apoptosis of H2O2‑induced osteoblasts. Parthenolide decreased the levels of reactive oxygen species, MDA, LDH and ALP. SOD and GPX levels were increased by parthenolide in H2O2‑induced osteoblasts. This suggested that parthenolide may break the equilibrium state of oxidative stress and inhibit cellular apoptosis. Parthenolide additionally increased the expression levels of oxidative stress‑associated genes, including nuclear factor erythroid 2 like 2, hemeoxygenase‑1 and quinone oxidoreductase 1 in H2O2‑induced osteoblasts. Furthermore, parthenolide increased the expression of osteogenesis‑associated genes, including runt‑related transcription factor 2, osteopontin, osteocalcin and collagen 1 in H2O2‑inducedosteoblasts. Therefore, it was concluded that parthenolide may be used in the treatment of osteoporosis.