Antioxidant ameliorates cisplatin-induced renal tubular cell death through inhibition of death receptor-mediated pathways.
We have recently demonstrated the direct involvement of the death receptor-mediated apoptotic pathways in cisplatin-induced renal tubular cell (RTC) death. Reactive oxygen species are thought to be a major cause of cellular damage in such injury. The aim of this study was to examine the mechanism through which antioxidants ameliorate cisplatin-induced RTC death, with special emphasis on death receptor-mediated apoptotic pathways. Cisplatin was added to cultures of normal rat kidney (NRK52E) cells or injected in rats. NRK52E cells and rats were also treated with dimethylthiourea (DMTU), a hydroxyl radical scavenger. We then examined the mRNA levels of death ligands and receptors, caspase-8 activity, cell viability, cell death, renal function, and histological alterations. RT-PCR indicated cisplatin-induced upregulation of Fas, Fas ligand, and TNF-alpha mRNAs and complete inhibition by DMTU in vitro and in vivo. Cisplatin increased caspase-8 activity of NRK52E cells, and DMTU prevented such activation. Exposure to cisplatin reduced viability of NRK52E cells, examined by WST-1 assay, and increased apoptosis and necrosis of the cells, examined by terminal deoxynucleotidyl transferase dUTP nick-end labeling assay and fluorescence-activated cell sorter analysis. DMTU abrogated cisplatin-induced changes in cell viability and apoptosis and/or necrosis. Cisplatin-induced renal dysfunction and histological damage were also prevented by DMTU. DMTU did not hinder cisplatin incorporation into RTCs. Our results suggest that antioxidants can ameliorate cisplatin-induced acute renal failure through inactivation of the death receptor-mediated apoptotic pathways.
Tsuruya
Kazuhiko
K
Department of Medicine and Clinical Science, Graduate School of Medical Sciences, Kyushu Univ., 3-1-1 Maidashi, Higashi-ku, Fukuoka 812-8582, Japan.
Tokumoto
Masanori
M
Ninomiya
Toshiharu
T
Hirakawa
Makoto
M
Masutani
Kohsuke
K
Taniguchi
Masatomo
M
Fukuda
Kyoichi
K
Kanai
Hidetoshi
H
Hirakata
Hideki
H
Iida
Mitsuo
M
D000818:Animals; D015703:Antigens, CD / Q000235:genetics; D000970:Antineoplastic Agents / Q000493:pharmacokinetics / Q000633:toxicity*; D000975:Antioxidants / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects*; D053181:Caspase 8; D053453:Caspase 9; D020169:Caspases / Q000378:metabolism; D002460:Cell Line; D002945:Cisplatin / Q000493:pharmacokinetics / Q000633:toxicity*; D053222:Fas Ligand Protein; D016166:Free Radical Scavengers / Q000494:pharmacology; D015870:Gene Expression / Q000187:drug effects; D066298:In Vitro Techniques; D007684:Kidney Tubules / Q000187:drug effects / Q000473:pathology* / Q000503:physiopathology; D008562:Membrane Glycoproteins / Q000235:genetics; D009336:Necrosis; D012333:RNA, Messenger / Q000032:analysis; D051381:Rats; D018124:Receptors, Tumor Necrosis Factor / Q000235:genetics; D047888:Receptors, Tumor Necrosis Factor, Type I; D013890:Thiourea / Q000031:analogs & derivatives* / Q000494:pharmacology*; D014409:Tumor Necrosis Factor-alpha / Q000235:genetics; D015854:Up-Regulation; D019014:fas Receptor / Q000235:genetics
D015703:Antigens, CD; D000970:Antineoplastic Agents; D000975:Antioxidants; D053222:Fas Ligand Protein; C505662:Faslg protein, rat; D016166:Free Radical Scavengers; D008562:Membrane Glycoproteins; D012333:RNA, Messenger; D018124:Receptors, Tumor Necrosis Factor; D047888:Receptors, Tumor Necrosis Factor, Type I; D014409:Tumor Necrosis Factor-alpha; D019014:fas Receptor; C038983:1,3-dimethylthiourea; C505528:Casp8 protein, rat; C506435:Casp9 protein, rat; D053181:Caspase 8; D053453:Caspase 9; D020169:Caspases; D013890:Thiourea; D002945:Cisplatin