RESULTS: We used the zebrafish model as well as in vitro cell-based assays to investigate AG-induced cell death and found that ferroptosis is the dominant type of cell death induced by neomycin. Neomycin stimulates lipid reactive oxygen species (ROS) accumulation through mitochondrial pathway and blocking mitochondrial ferroptosis pathway effectively protects neomycin-induced cell death. We screened an alkaloid natural compound library and identified seven small compounds that protect neomycin-induced ototoxicity by targeting ferroptosis pathway: six of them are radical-trapping agents (RTAs) while the other one (ellipticine) regulates intracellular iron homeostasis, which is essential for the generation of lipid ROS to stimulate ferroptosis.
CONCLUSIONS: Our study demonstrates that blocking intracellular ferroptosis pathway is an alternative strategy to ameliorate neomycin-induced ototoxicity and provides multiple hit compounds for further otoprotective drug development.
结果:我们使用斑马鱼模型以及体外基于细胞的测定法来研究AG诱导的细胞死亡,发现铁死亡是新霉素诱导的细胞死亡的主要类型。新霉素通过线粒体途径刺激脂质活性氧(ROS)积累,阻断线粒体铁凋亡途径有效保护新霉素诱导的细胞死亡。我们筛选了一个生物碱天然化合物库,并鉴定了7种通过靶向铁凋亡途径保护新霉素诱导的耳毒性的小化合物:其中6种是自由基捕获剂(RTAs),而另一种(椭圆碱)调节细胞内铁稳态,这对于脂质ROS的产生以刺激铁凋亡是必不可少的。
结论:我们的研究表明,阻断细胞内铁凋亡途径是改善新霉素诱导的耳毒性的替代策略,并为进一步的耳保护药物开发提供了多种化合物。