Overload of ferric ammonium citrate triggers MAPK pathways by producing high level of reactive oxygen species and induces apoptosis of human hFOB1.19 osteoblast cells

AIM:To investigate the role of reative oxygen species(ROS) generated by iron overload in activating the mitogen-activated protein kinase(MAPK) pathways and apoptosis.METHODS:Cultured human osteoblast cell line hFOB1.19 was treated with ferric ammonium citrate(FAC) at concentrations of 0-500 μmoL/L.The proliferation of hFOB1.19 cells was analyzed by MTT assay.Apoptosis was detected by flow cytometry with Annexin V/PI staining.The expression levels of p-ERK,p-JNK and p-p38 were determined by Western blotting 24 h after treatment with FAC.RESULTS:After treated with FAC,the cell proliferation was inhibited.The early apoptosis and total cell death were significantly increased.The levels of ROS were increased to(35.73±2.52)%,(62.89±4.24)% and(76.06±3.55)% with the increasing doses of FAC treatmen,respectively.The expression levels of p-ERK,p-JNK and p-p38 were also remarkably elevated in FAC groups.CONCLUSION:Iron overload increases intracellular ROS level,thus triggering the MAPK pathways and inducing apoptosis of human hFOB1.19 osteoblast cells.