The effects of Tempol on ferritin synthesis and Fe metabolism in lens epithelial cells

摘要

The nitroxide, Tempol, can protect tissue from oxidative damage by removing superoxide and by oxidizing Fe(II) to Fe(III), thus decreasing formation of the hydroxyl radical. However, long-term exposure to Tempol can damage cells. The oxidation of Fe could have profound effects on Fe metabolism in cells, yet this has not been previously studied. In the present investigation, the effects of Tempol on the synthesis of the Fe storage protein, ferritin, and its ability to store Fe were studied in cultured lens epithelial cells (LEC). In addition, the effects of short- and long-term Tempol treatment on the resistance of LEC to oxidative stress were determined. Tempol had a clear does-dependent inhibitory effect on ferritin synthesis noted at 6 h. By 20 h, ferritin synthesis returned toward normal levels. However, Fe incorporation into ferritin was decreased by almost 90% by the highest dose of Tempol, even at the 20-h time point. The decrease in Fe incorporation into ferritin was accompanied by a significant increase in the LMW pool of Fe. After short-term (4 h) treatment with Tempol, LEC were protected against the toxic effects of tertiary butyl hydroperoxide. However, after longer term treatment (20 h), Tempol itself had a toxic effect and did not afford protection. Indeed, at the higher doses, Tempol significantly reduced the ability of the cells to withstand oxidative stress. The redistribution of Fe within the cell after 20 h of Tempol treatment appears to render the cells more vulnerable to oxidative stress. The deleterious effects of Tempol on LEC are likely due to its effects on Fe metabolism, perhaps by reducing the availability of Fe for incorporation into ferritin and Fe-dependent enzymes as well as enlarging a low molecular weight pool of Fe which may be capable of catalyzing damaging free radical reactions.