Liproxstatin-1 Protects Hair Cell-Like HEI-OC1 Cells and Cochlear Hair Cells against Neomycin Ototoxicity
Zhiwei Zheng, Dongmei Tang, Liping Zhao, Wen Li, Jinghong Han, Bing Hu, Guohui Nie, Yingzi He
Abstract
Ferroptosis is a recently discovered iron-dependent form of oxidative programmed cell death distinct from caspase-dependent apoptosis. In this study, we investigated the effect of ferroptosis in neomycin-induced hair cell loss by using selective ferroptosis inhibitor liproxstatin-1 (Lip-1). Cell viability was identified by CCK8 assay. The levels of reactive oxygen species (ROS) were determined by DCFH-DA and cellROX green staining. The mitochondrial membrane potential ( <a:math xmlns:a="http://www.w3.org/1998/Math/MathML" id="M1"><a:mi>Δ</a:mi><a:mi>Ψ</a:mi><a:mi>m</a:mi></a:math> ) was evaluated by TMRM staining. Intracellular iron and lipid peroxides were detected with Mito-FerroGreen and Liperfluo probes. We found that ferroptosis can be induced in both HEI-OC1 cells and neonatal mouse cochlear explants, as evidenced by Mito-FerroGreen and Liperfluo staining. Further experiments showed that pretreatment with Lip-1 significantly alleviated neomycin-induced increased ROS generation and disruption in <c:math xmlns:c="http://www.w3.org/1998/Math/MathML" id="M2"><c:mi>Δ</c:mi><c:mi>Ψ</c:mi><c:mi>m</c:mi></c:math> in the HEI-OC1 cells. In parallel, Lip-1 significantly attenuated neomycin-induced hair cell damage in neonatal mouse cochlear explants. Collectively, these results suggest a novel mechanism for neomycin-induced ototoxicity and suggest that ferroptosis inhibition may be a new clinical intervention to prevent hearing loss.