Antioxidative Stress-Induced Destruction to Cochlear Cells Caused by Blind Antioxidant Therapy

Objective: Verification that blind and excessive use of antioxidants leads to antioxidant stress which exacerbates cochlear cell damage. Study Design: Basic research. Setting: The Third Affiliated Hospital of Sun Yat-Sen University. Methods: We compared and quantified hair cell-like house ear institute-organ of corti 1 (HEI-OC1) cell density, cell viability, and apoptosis caused by different concentrations of N-acetylcysteine (NAC) via Hoechst staining, Cell Counting Kit 8, Hoechst with propidium iodide staining, and Annexin V with propidium iodide (PI) staining. Apoptosis induced by high concentrations of M40403 and coenzyme Q10 in cochlear explants was analyzed and compared by cochlear dissection and activated caspase 3 labeling. Results: With the increase of NAC concentration (0-1000 μmol/L), cell density decreased consequently and reached the lowest at 1000 μmol/L (****P ≤.0001). Cell viability is also declining (**P <.01). The number of Annexin V-fluorescein isothiocyanate-labeled cells and PI-labeled cells increased with increasing NAC concentration after treatment of HEI-OC1 cells for 48 hours. The proportion of apoptotic cells also rose (*P <.05, **P <.01). Cochlear hair cells (HCs) treated with low concentrations of M40403 and coenzyme Q10 for 48 hours showed no damage. When the concentrations of M40403 and coenzyme Q10 were increased (concentrations＞30 μmol/L), HC damage began, followed by a dose-dependent increase in HC loss (*P <.001, **P <.0001). Activated caspase-3 was clearly apparent in cochlear explants treated with 50 μmol/L M40403 and coenzyme Q10 compared with cochlear explants without added M40403 and coenzyme Q10. Conclusion: These experimental results suggest that inappropriate application of antioxidants can cause severe damage to normal cochlear HCs.