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The miR-182-5p/GPX4 Pathway Contributes to Sevoflurane-Induced Ototoxicity via Ferroptosis
Lin Jin1, Xiaopei Yu2,3, Xuehua Zhou1
1Department of Anesthesiology, Eye and ENT Hospital, Fudan University, Shanghai 200031, China.
Abstract:
Our study aimed to investigate the role of ferroptosis in sevoflurane-induced hearing impairment and explore the mechanism of the microRNA-182-5p (miR-182-5p)/Glutathione Peroxidase 4 (GPX4) pathway in sevoflurane-induced ototoxicity. Immunofluorescence staining was performed using myosin 7a and CtBP2. Cell viability was assessed using the CCK-8 kit. Fe2+ concentration was measured using FerroOrange and Mi-to-FerroGreen fluorescent probes. The lipid peroxide level was assessed using BODIPY 581/591 C11 and MitoSOX fluorescent probes. The auditory brainstem response (ABR) test was conducted to evaluate the hearing status. Bioinformatics tools and dual luciferase gene reporter analysis were used to confirm the direct targeting of miR-182-5p on GPX4 mRNA. GPX4 and miR-182-5p expression in cells was assessed by qRT-PCR and Western blot. Ferrostatin-1 (Fer-1) pretreatment significantly improved hearing impairment and damage to ribbon synapses in mice caused by sevoflurane exposure. Immunofluorescence staining revealed that Fer-1 pretreatment reduced intracellular and mitochondrial iron overload, as well as lipid peroxide accumulation. Our findings indicated that miR-182-5p was upregulated in sevoflurane-exposed HEI-OC1 cells, and miR-182-5p regulated GPX4 expression by binding to the 3'UTR of GPX4 mRNA. The inhibition of miR-182-5p attenuated sevoflurane-induced iron overload and lipid peroxide accumulation. Our study elucidated that the miR-182-5p/GPX4 pathway was implicated in sevoflurane-induced ototoxicity by promoting ferroptosis.
Insights
Sevoflurane exposure causes hearing loss by promoting ferroptosis, a cell death process. The microRNA-182-5p (miR-182-5p) targets Glutathione Peroxidase 4 (GPX4), exacerbating this damage.
Area of Science:
- Ototoxicity and Neurobiology
- Cellular and Molecular Mechanisms
- Biomedical Research
Background:
- Anesthetic agents like sevoflurane can cause ototoxicity, leading to hearing impairment.
- Ferroptosis, a regulated form of cell death characterized by iron accumulation and lipid peroxidation, is implicated in various cellular damages.
- The specific molecular pathways underlying sevoflurane-induced ototoxicity require further elucidation.
Purpose of the Study:
- To investigate the role of ferroptosis in sevoflurane-induced hearing impairment.
- To explore the mechanism of the microRNA-182-5p (miR-182-5p)/Glutathione Peroxidase 4 (GPX4) pathway in sevoflurane-induced ototoxicity.
- To identify potential therapeutic targets for mitigating sevoflurane-induced hearing damage.
Main Methods:
- Auditory brainstem response (ABR) tests and immunofluorescence staining (myosin 7a, CtBP2) were used to assess hearing status and synapse integrity in mice.
- Cell viability (CCK-8), intracellular iron (FerroOrange, Mi-to-FerroGreen), and lipid peroxide levels (BODIPY 581/591 C11, MitoSOX) were measured in HEI-OC1 cells.
- Quantitative real-time PCR (qRT-PCR) and Western blot analyzed GPX4 and miR-182-5p expression.
- Bioinformatics and dual luciferase reporter assays confirmed the interaction between miR-182-5p and GPX4 mRNA.
Main Results:
- Ferrostatin-1 (Fer-1) pretreatment significantly ameliorated hearing impairment and ribbon synapse damage caused by sevoflurane, reducing iron overload and lipid peroxidation.
- Sevoflurane exposure upregulated miR-182-5p in HEI-OC1 cells.
- miR-182-5p directly targeted GPX4 mRNA, and its inhibition attenuated sevoflurane-induced ferroptosis.
- GPX4 expression was inversely regulated by miR-182-5p.
Conclusions:
- The miR-182-5p/GPX4 pathway plays a critical role in sevoflurane-induced ototoxicity by promoting ferroptosis.
- Inhibition of miR-182-5p may represent a therapeutic strategy to prevent or treat sevoflurane-induced hearing impairment.
- Understanding the ferroptosis mechanism provides insights into anesthetic-induced cellular damage.
