Clin Exp Otorhinolaryngol. 2014 Dec;7(4):286-94. doi: 10.3342/ceo.2014.7.4.286. Epub 2014 Nov 14.

Protective effect of metformin on gentamicin-induced vestibulotoxicity in rat primary cell culture.

Clinical and experimental otorhinolaryngology

Ji Young Lee, Se Hee Lee, Ji Won Chang, Jae-June Song, Hak Hyun Jung, Gi Jung Im

PMID: 25436048 PMCID: PMC4240486 DOI: 10.3342/ceo.2014.7.4.286

Abstract

OBJECTIVES: One of the antidiabetic drugs, metformin, have shown that it prevented oxidative stress-induced death in several cell types through a mechanism involving the opening of the permeability transition pore and cytochrome c release. Thus, it is possible that the antioxidative effect of metformin can also serve as protection against gentamicin-induced cytotoxicity related to reactive oxygen species (ROS). The aim of this study was to examine the protective effect of metformin on gentamicin-induced vestibulotoxicity in primary cell culture derived from rat utricle.

METHODS: For vestibular primary cell culture, rat utricles were dissected and incubated. Gentamicin-induced cytotoxicity was measured in both the auditory and vestibular cells. To examine the effects of metformin on gentamicin-induced cytotoxicity in the primary cell culture, the cells were pretreated with metformin at a concentration of 1 mM for 24 hours, and then exposed to 2.5 mM gentamicin for 48 hours. The intracellular ROS level was measured using a fluorescent dye, and also measured using a FACScan flow cytometer. Intracellular calcium levels in the vestibular cells were measured with calcium imaging using Fura-2 AM.

RESULTS: Vestibular cells were more sensitive to gentamicin-induced cytotoxicity than auditory hair cells. Metformin protects against gentamicin-induced cytotoxicity in vestibular cells. Metformin significantly reduced a gentamicin-induced increase in ROS, and also reduced an increase in intracellular calcium concentrations in gentamicin-induced cytotoxicity.

CONCLUSION: Metformin significantly reduced a gentamicin-induced increase in ROS, stabilized the intracellular calcium concentration, and inhibited gentamicin-induced apoptosis. Thus, Metformin showed protective effect on gentamicin-induced cytotoxicity in vestibular primary cell culture.

Keywords: Calcium; Gentamicins; Labyrinth vestibule; Metformin; Reactive oxygen species

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