Protective Effects of Anethole Dithiolethione against Oxidative Stress-induced Cytotoxicity in Human Jurkat T Cells

Abstract

The protective effects of anethole dithiolethione (ADT) against H 2O 2- or 4-hydroxynonenal (HNE)-induced cytotoxicity in human Jurkat T cells were investigated. Jurkat T cells were pretreated with ADT (10–50 μM) for 18 hr and then challenged with H 2O 2 or HNE for up to 4 hr. Cytotoxicity was assessed by measuring: 1) leakage of lactate dehydrogenase from cells to medium; and 2) exclusion of the DNA intercalating fluorescent probe propidium iodide by viable cells. Pretreatment of cells with ADT (10 or 25 μM) for 18 hr significantly protected cells against H 2O 2- or HNE-induced cytotoxicity. Treatment of cells with ADT (10–50 μM) for 72 hr significantly increased the activities of catalase and glutathione reductase. The maximum effect of ADT treatment on the activity of these enzymes was observed when cells were treated with 25 μM of ADT for 72 hr. A significant increase in cellular GSH was observed in cells that were treated with ADT for 72 hr. Using monobromobimane as a thiol probe, we consistently observed that cells pretreated for 18 hr with ADT (25 or 50 μM) had also increased total thiol content. Exposure of Jurkat T cells to H 2O 2 or HNE resulted in a time-dependent decrease in cellular GSH. ADT (10–50 μM, 18 hr) pretreatment circumvented H 2O 2-dependent lowering of cellular GSH. In conclusion, ADT proved to be a potent cytoprotective thiol antioxidant with multifaceted mechanisms of action, suggesting that the drug has a remarkable therapeutic potential.