Cephaloridine-induced inhibition of cytochrome c oxidase activity in the mitochondria of cultured renal epithelial cells (LLC-PK(1)) as a possible mechanism of its nephrotoxicity.
To clarify the mechanism of cephalosporin nephrotoxicity, the effects of cephaloridine (CLD), a nephrotoxic cephalosporin antibiotic, on the mitochondria of the pig kidney proximal tubular epithelial cell line LLC-PK(1) were studied in culture. The activity of cytochrome c oxidase in the mitochondria of LLC-PK(1) cells was significantly decreased from 9 h after addition of 1.0 mM CLD to the cultured cells. These effects were dose-dependent and accompanied with a significant decrease in the ATP content in the cells, followed by marked morphological changes in the mitochondria. These alterations were observed in the treated cells before the increase in lipid peroxidation. The activities of NADH-cytochrome c reductase and succinate dehydrogenase in the mitochondria and NADPH-cytochrome P450 reductase, NADH-cytochrome b(5) reductase, and 7-ethoxycoumarin O-deethylase in the microsomes of the treated cells were not affected. Superoxide anion production by the mitochondria prepared from LLC-PK(1) cells or NADH-cytochrome c reductase was not affected by addition of CLD (1-10 mM), but adriamycin (0.1 mM) or paraquat (0.1 mM) significantly increased the superoxide anion production. These results suggested that the primary action of CLD is inhibition of cytochrome c oxidase activity in the mitochondrial electron transport chain, which decreases intracellular ATP content in renal tubular epithelial cells and that these effects of CLD are followed by increased lipid peroxidation and cellular injury.
Kiyomiya
K
K
Department of Toxicology, School of Veterinary Medicine, Osaka Prefecture University, 1-1 Gakuen-cho, Sakai, Osaka, 599-8531, Japan. kiyomiyua@vet.osakafu-u.ac.jp
Matsushita
N
N
Matsuo
S
S
Kurebe
M
M
D000818:Animals; D002478:Cells, Cultured; D002509:Cephaloridine / Q000494:pharmacology*; D002511:Cephalosporins / Q000494:pharmacology*; D004317:Doxorubicin / Q000494:pharmacology; D003576:Electron Transport Complex IV / Q000037:antagonists & inhibitors*; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D004847:Epithelial Cells / Q000166:cytology / Q000187:drug effects* / Q000648:ultrastructure; D007674:Kidney Diseases / Q000139:chemically induced / Q000378:metabolism; D007687:Kidney Tubules, Proximal / Q000166:cytology / Q000187:drug effects; D015227:Lipid Peroxidation / Q000187:drug effects; D008861:Microsomes / Q000187:drug effects / Q000201:enzymology; D008928:Mitochondria / Q000000981:diagnostic imaging / Q000187:drug effects* / Q000201:enzymology; D010269:Paraquat / Q000494:pharmacology; D013552:Swine; D014463:Ultrasonography
D002511:Cephalosporins; D004791:Enzyme Inhibitors; D004317:Doxorubicin; D003576:Electron Transport Complex IV; D002509:Cephaloridine; D010269:Paraquat