Influence of glutathione on zinc-mediated cellular toxicity

The effect of zinc on various pulmonary cell lines has been studied by measuring the depletion of total cellular glutathione after exposure to zinc(II) chloride at different concentrations. Total cellular glutathione (cGS) was measured at 31+/-3 nmol/mg, 3.8+/-0.6 nmol/mg, and 3.7+/-1.2 nmol/mg protein in A549, L2, and 11Lu cells, respectively. After treatment with buthionine sulfoximine (BSO), the cGS levels decreased by 20% in A549 cells and below 0.2 nmol/mg in L2 and 11Lu cells. Exposure of A549 cells to 25-200 microM ZnCl2 for 4 h alone decreased the cGS content to 60-80%. There was little additional effect in BSO-pretreated cells. In L2 and 11Lu cells, the decrease of cGS was 70-85% following exposure to 15-150 microM ZnCl2 for 2 h. If BSO was also used, the decrease in cGS was 85-95% in L2 cells and 75-85% in 11Lu cells. Exposure to 25-250 microM ZnCl2 for 2 h diminished protein synthesis as determined by radiolabeled methionine incorporation, with half-maximum inhibition (EC50) from 40-160 microM ZnCl2. To attain similar EC50 values in BSO-pretreated cells, only about half the zinc concentrations were required as compared to cells without pretreatment. The decrease of cGS was accompanied by an increased ratio of oxidized: reduced glutathione that was more pronounced in cells with low glutathione content.