Role of poly(ADP-ribose)polymerase in cisplatin-induced injury in LLC-PK1 cells.
Acute renal failure is a dose-limiting factor during cisplatin chemotherapy. We have previously shown in rats that the hydroxyl radical scavenger edaravone reverses cisplatin-induced in vivo renal damage. In the present study, the role of poly(ADP-ribose) polymerase (PARP) in cisplatin nephrotoxicity was investigated in porcine tubular cells LLC-PK1. Cell injury was elicited by transient exposure to 500 microM cisplatin for 1 h or continuous exposure to 30 microM cisplatin for 24 h. Various hydroxyl radical scavengers reversed cell damage in a transient but not permanent model. The cell injury seemed to be necrosis and apoptosis in transient and permanent models, respectively, as assessed by TUNEL method and Annexin V stain. PARP inhibitors such as 3-aminobenzamide and benzamide inhibited cell damage in transient but not permanent model. PARP-dependent cell injury was also observed after transient exposure to hydroxyl radical-generating solution. We demonstrated for the first time the activation of PARP in renal tubular cells by transient cisplatin exposure, as determined by immunofluorescent stain with anti-poly(ADP-ribose) antibody. Moreover, ATP was depleted by transient exposure to cisplatin or hydroxyl radical, both of which were reversed by PARP inhibitors. These findings suggest that hydroxyl radical generation followed by PARP activation contributes to the necrotic cell injury caused by a transient exposure to cisplatin.
Shino
Yuki
Y
Department of Hospital Pharmacy, Faculty of Medicine, Kyushu University, Fukuoka, Japan.
Itoh
Yoshinori
Y
Kubota
Toshio
T
Yano
Takahisa
T
Sendo
Toshiaki
T
Oishi
Ryozo
R
D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D017304:Annexin A5 / Q000378:metabolism; D000970:Antineoplastic Agents / Q000633:toxicity*; D017209:Apoptosis / Q000187:drug effects*; D002945:Cisplatin / Q000633:toxicity*; D004791:Enzyme Inhibitors / Q000494:pharmacology; D016166:Free Radical Scavengers / Q000494:pharmacology; D017665:Hydroxyl Radical / Q000378:metabolism*; D020287:In Situ Nick-End Labeling; D007684:Kidney Tubules / Q000201:enzymology* / Q000473:pathology*; D018374:LLC-PK1 Cells; D009336:Necrosis; D000067856:Poly(ADP-ribose) Polymerase Inhibitors; D011065:Poly(ADP-ribose) Polymerases / Q000276:immunology / Q000378:metabolism*; D013552:Swine
D017304:Annexin A5; D000970:Antineoplastic Agents; D004791:Enzyme Inhibitors; D016166:Free Radical Scavengers; D000067856:Poly(ADP-ribose) Polymerase Inhibitors; D017665:Hydroxyl Radical; D000255:Adenosine Triphosphate; D011065:Poly(ADP-ribose) Polymerases; D002945:Cisplatin