Metallic nanoparticles have gained significant attention in the area of biomedical technology. Because of its high surface area, metallic nanoparticles are being widely used in various fields including the medical and engineering sciences. One of the valuable applications of metallic nanoparticles especially copper, zinc, and iron nanoparticles is increasing the physiological function of central nervous system. Besides, Iranian people are using the Salvia chloroleuca for neuroprotective properties. In the present research, iron nanoparticles were biosynthesized by S. chloroleuca leaf aqueous extract as reducing and stabilizing agents. Also, we revealed the protective effect of FeNPs in methadone‐treated PC12 cells. FeNPs were characterized and analyzed using common nanotechnology techniques including FT‐IR, UV–Vis. spectroscopy; EDS, TEM, and FE‐SEM. TEM and FE‐SEM images revealed a uniform spherical morphology for FeNPs. In the biological part of the current study, the both treatments of FeNPs significantly (p ≤ 0.01) reduced the cell cytotoxicity and cell death index as well as increased the cell viability and cell proliferation in methadone‐treated PC12 cells. In these treatments, mitochondrial membrane potential significantly (p ≤ 0.01) increased compared to methadone‐induced PC12 cells. DPPH free radical scavenging test was did to evaluate the antioxidant potentials of FeCl3, S. chloroleuca, and FeNPs. DPPH test indicated similar antioxidant activities for S. chloroleuca, FeNPs, and butylated hydroxytoluene. In current experiment, we concluded that iron nanoparticles biosynthesized by S. chloroleuca leaf aqueous extract suppressed methadone‐induced cell death in a dose‐dependent manner in PC12 cells.