Effect of iron and ascorbate on cyclosporine-induced oxidative damage of kidney mitochondria and microsomes.
The stimulatory effect of iron and ascorbate on the damaging action of cyclosporine in kidney mitochondria, microsomes and epithelial cells was examined. Cyclosporine induced malondialdehyde formation and hydrogen peroxide production in mitochondria and attenuated the activity of MnSOD and glutathione peroxidase. The damaging effect of cyclosporine (50 microM) plus Fe2+(20 microM) on mitochondrial and microsomal lipids and proteins as well as mitochondrial thiols was greater than the summation of the oxidizing action of cyclosporine alone and Fe2+ alone. As for tissue components, iron enhanced cyclosporine-induced viability loss in kidney epithelial cells. Fe2+, EDTA and H2O2- induced 2-alpha deoxyribose degradation was attenuated by 10 mM DMSO and 200 microM DTPA but not affected by 200 microM cyclosporine. The addition of Fe2+ caused a change in the absorbance spectrum of cyclosporine in the wavelength range 230-350 nm. The simultaneous addition of cyclosporine (50 microM) and ascorbate (100 microM) showed the enhanced peroxidative effect on mitochondrial and microsomal lipids, which was inhibited by DTPA and EDTA (1 mM). Similar to iron, ascorbate enhanced cyclosporine-induced cell viability loss. The results show that iron and ascorbate promote the damaging action of cyclosporine in kidney cortex mitochondria and microsomes and in kidney epithelial cells, which may contribute to the enhancement of cyclosporine-induced nephrotoxicity.
Department of Pharmacology, College of Medicine, Chung-Ang University, Seoul, 156-756, South Korea. leecs@cau.ac.kr
S H
SH
Lee
Y C
YC
Yoon
Y Y
YY
Jang
J H
JH
Song
E S
ES
Han
C S
CS
Lee
D000818:Animals; D000975:Antioxidants / Q000494:pharmacology*; D001205:Ascorbic Acid / Q000494:pharmacology*; D002460:Cell Line; D016572:Cyclosporine / Q000009:adverse effects*; D004791:Enzyme Inhibitors / Q000009:adverse effects*; D004847:Epithelial Cells / Q000187:drug effects / Q000201:enzymology; D005296:Ferrous Compounds / Q000494:pharmacology*; D007668:Kidney / Q000166:cytology / Q000187:drug effects* / Q000201:enzymology; D015227:Lipid Peroxidation / Q000187:drug effects / Q000502:physiology; D008297:Male; D051379:Mice; D008861:Microsomes / Q000187:drug effects* / Q000201:enzymology; D008928:Mitochondria / Q000187:drug effects* / Q000201:enzymology; D051381:Rats
D000975:Antioxidants; D004791:Enzyme Inhibitors; D005296:Ferrous Compounds; D016572:Cyclosporine; D001205:Ascorbic Acid