title: Necroptosis contributes to the cyclosporin A-induced cytotoxicity in NRK-52E cells.
abstract: Cyclosporin A (CsA) induces renal tubular epithelial cells apoptosis and necrosis following in vitro exposure. The mechanisms of CsA-induced apoptosis have been studied intensively, whereas the mechanisms of necrosis remain to be elucidated. Necroptosis has been described as programmed necrosis. This study investigated the ability of CsA to induce necroptosis in the rat tubular cell line NRK-52E. The NRK-52E cells were incubated with CsA for 24 hours with or without necrostatin-1 (Nec-1). The majority of the NRK-52E cells died of necrosis as indicated by LDH leakage, Hoechst 33342/PI staining, and flow cytometry analysis. Cell death was significantly reduced by Nec-1 pretreated before CsA exposure. CsA-induced apoptosis and necrosis were also compared in NRK-52E cells with or without knockdown of receptor interaction protein 3 (RIP3) expression using small interfering RNA. Moreover, the role of reactive oxygen species (ROS) in CsA-induced cell death was also attempted. The result suggests that necroptosis contributes to the CsA-induced cytotoxicity in NRK-52E cells. Meanwhile, RIP3 and ROS are involved in CsA-induced necroptosis. To our knowledge, this is the first report on necroptosis in CsA-induced renal tubular cell death pathways, which might offer a novel protective target for CsA nephrotoxicity.
authors.affiliation: School of Pharmaceutical Sciences, Sun Yat-sen University, Guangzhou, PR China.
authors.forename: Zizhang
authors.initials: Z
authors.lastname: Ouyang
authors.forename: Shaohua
authors.initials: S
authors.lastname: Zhu
authors.forename: Jing
authors.initials: J
authors.lastname: Jin
authors.forename: Jia
authors.initials: J
authors.lastname: Li
authors.forename: Yuwen
authors.initials: Y
authors.lastname: Qiu
authors.forename: Min
authors.initials: M
authors.lastname: Huang
authors.forename: Zhiying
authors.initials: Z
authors.lastname: Huang
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000187:drug effects*; D001562:Benzimidazoles; D015153:Blotting, Western; D016923:Cell Death / Q000187:drug effects; D002460:Cell Line; D016572:Cyclosporine / Q000633:toxicity*; D005434:Flow Cytometry; D007093:Imidazoles / Q000378:metabolism; D007166:Immunosuppressive Agents / Q000633:toxicity*; D007211:Indoles / Q000378:metabolism; D007683:Kidney Tubular Necrosis, Acute / Q000139:chemically induced / Q000473:pathology*; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D009336:Necrosis / Q000139:chemically induced* / Q000473:pathology; D034741:RNA, Small Interfering / Q000494:pharmacology; D051381:Rats; D017382:Reactive Oxygen Species / Q000378:metabolism
chemical_list: D001562:Benzimidazoles; D007093:Imidazoles; D007166:Immunosuppressive Agents; D007211:Indoles; D034741:RNA, Small Interfering; D017382:Reactive Oxygen Species; C507699:necrostatin-1; D016572:Cyclosporine; D007770:L-Lactate Dehydrogenase; C017807:bisbenzimide ethoxide trihydrochloride