Effect of early acute high concentrations of iodide exposure on mitochondrial superoxide production in FRTL cells

Abstract Excessive oxidative stress has been suggested as one of the underlying mechanisms in the development of thyroid cytotoxicity. Although the involvement of mitochondria has been hypothesized, the effect of early acute high concentrations of iodide on mitochondrial superoxide production remains largely unknown, especially within a 24 h time frame. By using a novel fluorescent probe, MitoSOX Red, we demonstrated the concentration response and time-course response of KI-induced mitochondrial superoxide production in the Fischer rat thyroid cell line (FRTL). A strong increase of MitoSOX Red fluorescence intensity in FRTL cells can be seen at 2 h following high concentrations of iodide exposure. Besides, we indicated that 6-propyl-2-thiouracil (PTU, 300 μM), thyroid-stimulating hormone (TSH, 10 mU/ml), and perchlorate (KClO 4 , 30 μM) can inhibit excessive iodide-induced strong mitochondrial superoxide production; however, diethyldithiocarbamic acid (DETC, 2 mM) can further increase excessive iodide-induced mitochondrial superoxide production. By using transmission electron microscopy (TEM), we noted accumulated myelinoid bodies with lipid droplets and numerous apoptotic nuclear bodies at 24 h in FRTL cells. In addition, we demonstrated a significant decrease in cytochrome c (cyt c) content in the mitochondria by enzyme linked immunosorbent assay (ELISA), and DNA fragments and significant increases in lactate dehydrogenase (LDH) activity were detected. We propose a sequence of events mediated by a strong mitochondrial superoxide production at 2 h, followed by lipid peroxidation, cell membrane damage with significant cyt c release, culminating in DNA fragmentation and apoptotic nuclear formation at 24 h, which may partly contribute to the underlying mechanisms of early acute iodide excess.