Effect of oxypurinol on cyclosporine toxicity in cultured EA, LLC-PK1 and MDCK cells

Abstract

It has been proposed that cyclosporin (CsA) toxicity may in part be due to the action of oxygen-based freee radicals. We compared the response of cultured endothelial (EA) and epithelial (LLC-PK1 and MDCK) cells to CsA, 250 or 1000 ng/mL for 24 or 72 h, with or without the xanthine oxidase inhibitor oxypurinol (Oxy), 10 mg/mL. CsA-induced alterations were seen on phase contrast and electron microscopy. In EA cells, swollen mitochondria and large cytoplasmic vacuoles surrounded by a thin membrane containing ribosomes were present at the periphery of the cell. In LLC-PK1 cells no vacuoles were present while in MDCK cells, the vacuoles were smaller and more centrally located. In both epithelial cell lines, mitochondria were distorted with loss of cristae.In all three cell lines, CsA depressed cell counts and decreased 3H-thymidine incorporation. Also, LDH release was accelerated. The addition of Oxy minimized the morphologic effect of CsA on all three cell lines with the effect more apparent in EA cells. The CsA-induced reduction of cell replication and increase in LDH release were also attenuated byOxy. These results support the notion that the peroxidative properties of CsA may be responsible in part for CsA-induced nephrotoxicity.