OBJECTIVE In view of the antioxidant role of ascorbic acid and the glutathione redox cycle in the lens, the authors have studied the relationship of the cycle to reduction of the oxidized product of ascorbic acid, dehydroascorbic acid (DHA), in lens epithelium. METHODS Cultured dog lens epithelial cells and intact rabbit lenses were exposed to various concentrations of DHA in experiments performed at 20 degrees C to minimize hydrolysis of the compound (t1/2 of 5 minutes at 37 degrees C). Levels of glutathione (GSH) and oxidized glutathione (GSSG) were measured in lens cells and whole lens epithelial by electrochemical detection. RESULTS Treatment of lens cells with 1 mM DHA for 0.5 to 3 hours in the absence of glucose (glucose is required for the reduction of GSSG through the glutathione redox cycle) produced from 60% to complete oxidation of GSH (controls contained negligible GSSG) and distinct morphologic changes (cell contraction and blebbing), as shown by scanning electron microscopy. Glucose prevented these effects and allowed nearly immediate recovery of GSH after DHA exposure in the absence of glucose. A dose-dependent response was observed for the formation of GSSG in cultured cells from 0.05 to 0.5 mM DHA in the absence of glucose. The results of experiments performed with DHA plus an inhibitor of glutathione reductase mimicked those obtained using DHA minus glucose. DHA produced a 3- to 10-fold stimulation of hexose monophosphate shunt activity in cultured lens cells and whole lenses, which was prevented by the inhibition of glutathione reductase. Treatment of whole lenses with DHA minus glucose also produced oxidation of epithelial GSH and was accompanied by the loss of lens transparency. No evidence was found for dehydroascorbate reductase activity in the lens epithelium. CONCLUSIONS The exposure of lenses and lens epithelial cells to DHA under conditions in which the glutathione redox cycle was compromised resulted in the disappearance of GSH in the tissues and the appearance of GSSG. The reduction of DHA was shown to be linked to the glutathione redox cycle by a nonenzymatic interaction between GSH and DHA. Reduction of DHA in the lens is important because of the potential toxicity of this oxidant and/or its degradation products.