Inhibition of ferroptosis protects House Ear Institute-Organ of Corti 1 cells and cochlear hair cells from cisplatin-induced ototoxicity

J Cell Mol Med. 2020 Oct;24(20):12065-12081. doi: 10.1111/jcmm.15839. Epub 2020 Sep 14.

Abstract

Ferroptosis is a recently recognized form of non-apoptotic cell death caused by an iron-dependent accumulation of lipid hydroperoxides, which plays important roles in a wide spectrum of pathological conditions. The present study was aimed to investigate the impact of ferroptosis on cisplatin-induced sensory hair cell damage. Cell viability was determined by Cell Counting Kit-8 and lactase dehydrogenase assays. The reactive oxygen species (ROS) levels were evaluated by 2,7-Dichlorodi-hydrofluorescein diacetate (DCFH-DA) and MitoSox-Red staining. Mitochondrial membrane potential (MMP) was measured by tetramethylrhodamine methyl ester (TMRM) staining. Lipid peroxidation, intracellular and mitochondrial iron were detected by Liperfluo, C11-BODIPY581/591 , FerroOrange and Mito-FerroGreen, respectively. We found that cisplatin treatment not only markedly augmented ROS accumulation, decreased the MMP, but increased lipid peroxidation and iron accumulation in House Ear Institute-Organ of Corti 1 (HEI-OC1) cells. Of note, treatment with the specific ferroptosis inhibitor ferrostatin-1 could effectively abrogate the cisplatin-induced toxicity and subsequent cell death. Specifically, the improvement of mitochondrial functions is important mechanisms for protective action of ferroptosis inhibitor against cisplatin-induced damages in HEI-OC1 cells. Moreover, inhibition of ferroptosis significantly protected murine cochlear hair cells against cisplatin damage. In addition, treatment murine cochlear hair cells with ferroptosis inducer, RSL3, significantly exacerbated cisplatin-induced damage, which could be alleviated by ROS inhibitor N-acetyl-L-cysteine. Collectively, our study indicated that ferroptosis inhibition could alleviate the cisplatin-induced ototoxicity via inactivation of lipid peroxide radical and improvement of mitochondrial function in hair cells.

Keywords: cisplatin; ferroptosis; mitochondrial function; ototoxicity; reactive oxygen species.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Aldehydes / metabolism
  • Animals
  • Carbolines / pharmacology
  • Cell Line
  • Cell Survival / drug effects
  • Cisplatin / adverse effects*
  • Cyclohexylamines / pharmacology
  • Cytoprotection* / drug effects
  • Ferroptosis* / drug effects
  • Hair Cells, Auditory / drug effects
  • Hair Cells, Auditory / pathology*
  • Iron / metabolism
  • Iron Overload / pathology
  • Membrane Potential, Mitochondrial / drug effects
  • Mice, Inbred C57BL
  • Ototoxicity / pathology*
  • Phenylenediamines / pharmacology
  • Reactive Oxygen Species / metabolism

Substances

  • Aldehydes
  • Carbolines
  • Cyclohexylamines
  • Phenylenediamines
  • RSL3 compound
  • Reactive Oxygen Species
  • ferrostatin-1
  • Iron
  • 4-hydroxy-2-nonenal
  • Cisplatin