Lipid peroxidation contributes to hydrogen peroxide induced cytotoxicity in renal epithelial cells

• Published in: Kidney international Vol. 49; no. 1; pp. 88 - 93
• Main Authors: Sheridan, Alice M., Fitzpatrick, Sean, Wang, Candice, Wheeler, David C., Lieberthal, Wilfred
• Format: Journal Article
• Language: English
• Published: New York, NY Elsevier Inc 01.01.1996; Nature Publishing
• Subjects: Animals; Biological and medical sciences; Cell Division - drug effects; Cell Membrane - drug effects; Cell Membrane - metabolism; Cells, Cultured; Epithelium - drug effects; Epithelium - metabolism; Epithelium - pathology; Fatty Acids, Unsaturated - metabolism; Hydrogen Peroxide - toxicity; Kidney Tubules, Proximal - drug effects; Kidney Tubules, Proximal - metabolism; Kidney Tubules, Proximal - pathology; Lipid Peroxidation; Malondialdehyde - metabolism; Medical sciences; Mice; Mice, Inbred C57BL; Nephrology. Urinary tract diseases; Nephropathies. Renovascular diseases. Renal failure; Oxidants - toxicity; Tubulopathies; Oxygen; Rodentia; Polyunsaturated fatty acid; Lipids; Cytotoxicity; Hydrogen Peroxides; In vitro; Kidney; Vertebrata; Renal tubule; Mammalia; Urinary system; Mouse; Cell death; Animal; Tubulointerstitial nephritis; Established cell line; Plasma membrane; Epithelial cell; Peroxidation
• ISSN: 0085-2538; 1523-1755
• DOI: 10.1038/ki.1996.12

Lipid peroxidation contributes to hydrogen peroxide induced cytotoxicity in renal epithelial cells. We have examined the role of lipid peroxidation in the cytotoxicity of H2O2 in OK cells containing markedly differing amounts of cell membrane polyunsaturated fatty acids (PUFA). In OK cells grown in a serum free medium, PUFA were undetectable. The membranes of these cells contained predominantly oleic, stearic and palmitic acids. When cultured in medium containing 10% calf serum, OK cells contained measurable amounts of PUFA [linoleic (5 ± 1%) and arachidonic acids (8 ± 1%)]. When the serum containing medium was supplemented with 60mm linoleic acid, the membrane content of both linoleic (21 ± 1%) as well as arachidonic acid (15 ± 1%) was substantially increased. The severity of injury induced by H2O2 in OK cells was substantially altered by the PUFA content of the cell membrane. Exposure of OK cells to 1.25mm H2O2 for one hour resulted in more cell death (determined by a trypan blue assay) in cells grown in serum supplemented with linoleic acid with “normal” PUFA content (90 ± 2%) than in cells with “reduced” levels of PUFA grown in unsupplemented calf serum (81 ± 3%). Cells grown in defined, serum free medium with undetectable levels of PUFA suffered the least H2O2-induced lethal cell injury (47 ± 8%). Comparable differences in the cytotoxicity of H2O2 among cells with differing PUFA content were found using a clonogenic assay of cell viability. Malondialdehyde (MDA) accumulation induced by 1.25mm H2O2 was greater in cells with “normal” PUFA content (702 ± 103 pm/µg cell DNA/hr) than in cells with “reduced” PUFA (328 ± 112 pm/100 µg DNA/hr) and was undetectable in cells grown in defined, serum free medium. In summary, the content of PUFA of cells in culture is profoundly influenced by culture conditions. Our data provide novel and direct evidence that peroxidation of cell membranes contributes directly to the severity of cell injury and death induced by H2O2.