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J Am Soc Nephrol 15:S85-S87, 2004
© 2004 American Society of Nephrology

Supplement Article

Oxidative Stress Mediates Apoptotic Changes Induced by Hyperglycemia in Human Tubular Kidney Cells

Daniela Verzola*, Maria Bianca Bertolotto{dagger}, Barbara Villaggio*, Luciano Ottonello{dagger}, Franco Dallegri{dagger}, Francesca Salvatore*, Valeria Berruti*, Maria Teresa Gandolfo*, Giacomo Garibotto* and Giacomo Deferrari*

*Division of Nephrology, Department of Internal Medicine, University of Genoa, Genoa, Italy; and {dagger}Division of Internal Medicine Department of Internal Medicine, University of Genoa, Genoa, Italy

Correspondence to Dr. Giacomo Garibotto, Department of Internal Medicine, Nephrology Division, Viale Benedetto XV 6, 16132 Genoa, Italy. Phone: ++390103538989; Fax: ++390103538959; E-mail: gari{at}unige.it

ABSTRACT. Reactive oxygen species (ROS) are important mediators for several biologic responses, including apoptosis. The present study evaluated the time course of changes in intracellular ROS production and apoptosis-related proteins, as well as apoptotic changes in human tubular proximal cells (HK-2 cells) exposed to hyperglycemia. Apoptosis (annexin V binding), ROS formation (fluorescence probe dichlorofluorescin diacetate and FACScan flow cytometry), and X chromosome–linked protein (XIAP; Western blot) were studied in HK-2 cells grown in a medium containing normal (NG) or high glucose (HG) concentrations (5.5 or 30 mM, respectively) for 18 to 48 h. HG promoted an increase (65% at 18 h and 73% at 24 h; P < 0.05 versus NG) in intracellular ROS generation. At 18 h, the NF-kB binding activity (evaluated by electrophoretic mobility-shift assay) was suppressed by HG. At the same time, the expression of NF-kB–induced antiapoptotic XIAP was reduced in HG-treated cells. Apoptotic changes were observed at 48 h (34 ± 7% in HG versus 10 ± 3% in NG; P < 0.001). Changes in ROS production at 24 h predicted changes in the apoptotic index at 48 h (r = 0.96, P < 0.0001). These results suggest that hyperglycemia induces apoptotic changes in human tubular cells via an increase in oxidative stress and that a downregulation of antiapoptotic protein XIAP is a component of this response.




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