Protective Effect of Metformin on Gentamicin-Induced Vestibulotoxicity in Rat Primary Cell Culture.
- Author:
Ji Young LEE
1
;
Se Hee LEE
;
Ji Won CHANG
;
Jae June SONG
;
Hak Hyun JUNG
;
Gi Jung IM
Author Information
- Publication Type:Original Article
- Keywords: Labyrinth vestibule; Gentamicins; Metformin; Reactive oxygen species; Calcium
- MeSH: Animals; Apoptosis; Calcium; Cytochromes c; Fura-2; Gentamicins; Hair Cells, Auditory; Hypoglycemic Agents; Metformin*; Permeability; Primary Cell Culture*; Rats*; Reactive Oxygen Species; Saccule and Utricle; Vestibule, Labyrinth
- From:Clinical and Experimental Otorhinolaryngology 2014;7(4):286-294
- CountryRepublic of Korea
- Language:English
- 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.
