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Am J Physiol Lung Cell Mol Physiol 275: L80-L86, 1998;
1040-0605/98 $5.00
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Vol. 275, Issue 1, L80-L86, July 1998

Differential regulation of glutathione by oxidants and dexamethasone in alveolar epithelial cells

Irfan Rahman1, Agnes Bel1, Brigitte Mulier1, Kenneth Donaldson2, and William MacNee1

1 The Rayne Laboratory, Respiratory Medicine Unit, Department of Medicine, University of Edinburgh, Edinburgh EH8 9AG; and 2 Department of Biological Sciences, Napier University, Edinburgh EH10 5DT, United Kingdom

We studied the regulation of GSH and the enzymes involved in GSH regulation, gamma -glutamylcysteine synthetase (gamma -GCS) and gamma -glutamyl transpeptidase (gamma -GT), in response to the oxidants menadione, xanthine/xanthine oxidase, hyperoxia, and cigarette smoke condensate in human alveolar epithelial cells (A549). Menadione (100 µM), xanthine/xanthine oxidase (50 µM/10 mU), and cigarette smoke condensate (10%) exposure produced increased GSH levels (240 ± 6, 202 ± 12, and 191 ± 2 nmol/mg protein, respectively; P < 0.001) compared with the control level (132 ± 8 nmol/mg protein), which were associated with a significant increase in gamma -GCS activity (0.18 ± 0.006, 0.16 ± 0.01, and 0.17 ± 0.008 U/mg protein, respectively; P < 0.01) compared with the control level (0.08 ± 0.001 U/mg protein) at 24 h. Exposure to hyperoxia (95% O2) resulted in a time-dependent increase in GSH levels. gamma -GCS activity increased significantly at 4 h (P < 0.001), returning to control values after 12 h of exposure. Dexamethasone (3 µM) exposure produced a significant time-dependent decrease in the levels of GSH and gamma -GCS activity at 24-96 h. The activity of gamma -GT did not change after oxidant treatment; however, it was decreased significantly by dexamethasone at 24-96 h. Thus oxidants and dexamethasone modulate GSH levels and activities of gamma -GT and gamma -GCS by different mechanisms. We suggest that the increase in gamma -GCS activity but not in gamma -GT activity may be required for the increase in intracellular GSH under oxidative stress in alveolar epithelial cells.

gamma -glutamylcysteine synthetase; gamma -glutamyl transpeptidase; A549 cells


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