Protective effect of apigenin against oxidative stress-induced damage in osteoblastic cells

Apigenin, a plant-derived flavonoid, was investigated to determine whether it could influence hydrogen peroxide (H2O2)-induced oxidative damage and cellular dysfunction in the MC3T3-E1 mouse osteoblastic cell line. In the present study, osteoblastic cells were treated with H2O2 in the presence or absence of apigenin. Cell viability, apoptosis, reactive oxygen species (ROS) production and mitochondrial membrane potential (Delta Psi m) were subsequently examined. It was observed that H2O2 reduced cell survival and Delta Psi m, while it markedly increased the intracellular levels of ROS and apoptosis. However, pretreatment of cells with apigenin attenuated all the H2O2-induced effects. The antioxidants, catalase and N-acetyl-L-cysteine (NAC) also prevented H2O2-induced oxidative cell damage. In addition, treatment with apigenin resulted in a significant elevation of osteoblast differentiation genes including alkaline phosphatase (ALP), collagen, osteopontin (OPN), osteoprotegerin (OPG), bone sialoprotein (BSP), osterix (OSX) and osteocalcin (OC) and bone morphogenetic proteins (BMPs) genes (BMP2, BMP4 and BMP7). In the mechanistic studies of cell signaling by the antioxidative potential of apigenin, it was found that apigenin activated the H2O2-induced decreased expression of phosphatidylinositol 3'-kinase (PI3K), protein kinase B2 (AKT2) genes and extracellular signal-related kinase (EPK) 2, which are key regulators of survival-related signaling pathways. By contrast, there were no changes in the expression of nuclear facor-kappa B (NF-kappa B) and c-Jun N-terminal kinase (JNK) gene exposed to H2O2 in the present study. Apigenin also upregulated the gene expression of antioxidant enzymes, superoxide dismutase (SOD) 1, SOD2 and glutathione peroxidase (GPx) 1. Taken together, these results suggested that apigenin attenuated oxidative-induced cell damage in osteoblastic cells and may be useful for the treatment of oxidative-related bone disease.