Efflux of glutathione and glutathione complexes from human erythrocytes in response to vanadate

• Published in: Blood cells, molecules, & diseases Vol. 50; no. 1; pp. 1 - 7
• Main Authors: Cakir, Yeliz, Yildiz, Deniz
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 01-01-2013
• Subjects: 4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid - pharmacology; Adenosine - analogs & derivatives; Adenosine - pharmacology; Biological Transport - drug effects; Cells, Cultured; Chloride-Bicarbonate Antiporters - antagonists & inhibitors; Chloride-Bicarbonate Antiporters - metabolism; Dose-Response Relationship, Drug; Erythrocytes - cytology; Erythrocytes - drug effects; Erythrocytes - metabolism; Glutathione - metabolism; Glutathione efflux; Human erythrocytes; Humans; L-Lactate Dehydrogenase - metabolism; Membrane transport; Methylation - drug effects; Multidrug Resistance-Associated Proteins - antagonists & inhibitors; Multidrug Resistance-Associated Proteins - metabolism; Propionates - pharmacology; Quinolines - pharmacology; Succimer - pharmacology; Sulfhydryl Compounds - metabolism; Vanadate; Vanadates - pharmacology; Vanadate; Human erythrocytes; Membrane transport; Glutathione efflux
• ISSN: 1079-9796; 1096-0961
• DOI: 10.1016/j.bcmd.2012.07.001

The main objective of the present study was to investigate if vanadate is extruded from the cells in a glutathione dependent manner resulting in the appearance of extracellular glutathione and complexes of glutathione with vanadium. Vanadate significantly depleted intracellular non-protein sulfhydryl (NPSH) levels in a time- and concentration-dependent manner. The intracellular NPSH level was decreased to 0.0±0.0μmol/ml erythrocyte when exposed to 10mM of vanadate for 4h. Extracellular NPSH level was increased concomitantly with the intracellular decrease and reached to 0.1410±0.005μmol/ml erythrocyte in 4h. Intracellular decrease and extracellular increase in NPSH levels were significantly inhibited in the presence of DIDS, a chloride-bicarbonate exchanger which also mediates phosphate and arsenate transport in erythrocytes. In parallel with the increase in extracellular NPSH levels, significant increases in extracellular glutathione levels were detected following exposure to vanadate. Extracellular glutathione levels reached to 0.0150±0.0.001, 0.0330±0.001, and 0.0576±0.002μmol/ml erythrocyte with 1, 5, and 10mM of vanadate respectively. Dimercaptosuccinic acid treatment of supernatants significantly increased the glutathione levels measured in the extracellular media. Utilization of MK571 an MRP inhibitor decreased the rate of glutathione efflux from erythrocytes suggesting a role for this membrane transporter in the process. A known methylation inhibitor periodate oxidized adenosine decreased the rate of glutathione efflux from erythrocytes. This observed decrease in extracellular GSH levels suggests that GSH release partly requires a proper cellular methylation process and that part of GSH detected in the extracellular media may arise from GSH–vandium complexes. The results of the present study indicate that human erythrocyte efflux glutathione in reduced free form and in conjugated form/s that can be recovered with dimercaptosuccinic acid when exposed to vanadate.