title: Synergistic effect of apoptosis and necroptosis inhibitors in cisplatin-induced nephrotoxicity.
abstract: Necroptosis is a nonapoptotic cell death pathway. We aim to study the effect of necrostatin-1 (a specific necroptosis inhibitor) in cisplatin-induced injury. We analyzed the effect of the combined use of inhibitors of apoptosis (z-vad) and necroptosis (necrostatin-1) in acute kidney injury by cisplatin in human proximal tubule cells. Our results showed moderate effectiveness in cytoprotection after treatment with z-vad. But the concomitant use of inhibitors (z-vad and necrostatin-1) presented synergistic and additive protection. The present study analyzed the caspase-3 activity and we observed a significant decrease in the group treated with z-vad and cisplatin. However we did not observe changes in the group treated with both inhibitors (z-vad and necrostatin-1) and cisplatin. Thus, demonstrating that necroptosis is a caspase-independent mechanism. We also analyzed the effect of necrostatin-1 in vivo model. C57BL/6 mice were treated with cisplatin and/or inhibitors. The concomitant use of inhibitors (z-vad and necrostatin-1) recovered renal function and decreased levels of urinary Ngal. Additionally, we analyzed the expression of RIP-1, a specific marker for necroptosis. In animals treated with cisplatin and z-VAD levels of RIP-1 were higher. This result reinforces that necroptosis occurs only in conditions where apoptosis was blocked. However, the use of both inhibitors (z-vad and necrostatin-1) provided additional protection. In conclusion, our study has a significant potential to show in vitro and in vivo protection obtained by necrostatin-1. Therefore, our results suggest that necroptosis may be an important mechanism of cell death after kidney injury.
authors.affiliation: Federal University of São Paulo, Rua Pedro de Toledo, 740, 2º andar, São Paulo, Brazil. vivian.tristao@bol.com.br.
authors.forename: Vivian Regina
authors.initials: VR
authors.lastname: Tristão
authors.affiliation: Federal University of São Paulo, Rua Pedro de Toledo, 740, 2º andar, São Paulo, Brazil.
authors.forename: Edson A
authors.initials: EA
authors.lastname: Pessoa
authors.affiliation: Federal University of São Paulo, Rua Pedro de Toledo, 740, 2º andar, São Paulo, Brazil.
authors.forename: Renata
authors.initials: R
authors.lastname: Nakamichi
authors.affiliation: Federal University of São Paulo, Rua Pedro de Toledo, 740, 2º andar, São Paulo, Brazil.
authors.forename: Luciana A
authors.initials: LA
authors.lastname: Reis
authors.affiliation: Federal University of São Paulo, Rua Pedro de Toledo, 740, 2º andar, São Paulo, Brazil.
authors.forename: Marcelo Costa
authors.initials: MC
authors.lastname: Batista
authors.affiliation: Federal University of São Paulo, Rua Pedro de Toledo, 740, 2º andar, São Paulo, Brazil.
authors.forename: Marcelino de Souza
authors.initials: Mde S
authors.lastname: Durão Junior
authors.affiliation: Federal University of São Paulo, Rua Pedro de Toledo, 740, 2º andar, São Paulo, Brazil.
authors.forename: Júlio Cesar Martins
authors.initials: JC
authors.lastname: Monte
mesh_terms: D058186:Acute Kidney Injury / Q000139:chemically induced / Q000235:genetics* / Q000473:pathology / Q000517:prevention & control*; D000209:Acute-Phase Proteins / Q000235:genetics / Q000652:urine; D000818:Animals; D001806:Blood Urea Nitrogen; D053148:Caspase 3 / Q000235:genetics / Q000378:metabolism; D016923:Cell Death / Q000187:drug effects; D002460:Cell Line; D002945:Cisplatin / Q000633:toxicity; D003404:Creatinine / Q000097:blood; D019610:Cytoprotection / Q000235:genetics; D004357:Drug Synergism; D020690:GTPase-Activating Proteins / Q000235:genetics / Q000378:metabolism; D005786:Gene Expression Regulation; D006801:Humans; D007093:Imidazoles / Q000494:pharmacology*; D007211:Indoles / Q000494:pharmacology*; D007677:Kidney Function Tests; D007687:Kidney Tubules, Proximal / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D000071068:Lipocalin-2; D054834:Lipocalins / Q000235:genetics / Q000652:urine; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D009842:Oligopeptides / Q000494:pharmacology*; D015513:Oncogene Proteins / Q000235:genetics / Q000652:urine; D015398:Signal Transduction
chemical_list: D000209:Acute-Phase Proteins; D020690:GTPase-Activating Proteins; D007093:Imidazoles; D007211:Indoles; D000071068:Lipocalin-2; D054834:Lipocalins; D009842:Oligopeptides; D015513:Oncogene Proteins; C490620:Ralbp1 protein, mouse; C517629:benzyloxycarbonyl-valyl-alanyl-aspartic acid; C507699:necrostatin-1; C071332:Lcn2 protein, mouse; D003404:Creatinine; C505409:Casp3 protein, mouse; D053148:Caspase 3; D002945:Cisplatin
keywords: Acute kidney injury; Cisplatin; Cytoprotection; Necroptosis; Necrostatin-1