Assessment of Mitochondrial Membrane Potential in HEI-OC1 and LLC-PK1 Cells Treated with Gentamicin and Mitoquinone

Abstract

Objective

To determine the effects of concurrent treatment with gentamicin and the mitochondria-targeted antioxidant mitoquinone (MitoQ; which may prevent gentamicin ototoxicity) on change in the mitochondrial membrane potential (Δψ_m), a precursor of apoptosis.

Study Design

Prospective and controlled.

Setting

Academic research laboratory.

Subjects and Methods

LLC-PK1 (Lilly Laboratories Culture–Pig Kidney Type 1) and HEI-OC1 (House Ear Institute Organ of Corti 1) cells—renal and auditory cell lines, respectively—were used in this study. Δψ_m was assessed by flow cytometry through the MitoProbe JC-1 Kit for Flow Cytometry in untreated LLC-PK1 and HEI-OC1 cells and cells exposed to low- (100µM) or high- (2000µM) dose gentamicin for 24 hours, with and without 0.5µM each of MitoQ or idebenone (IDB; an untargeted ubiquinone).

Results

Δψ_m was not different in untreated LLC-PK1 cells and cells coincubated with low-dose gentamicin and MitoQ or IDB (P > .05). In HEI-OC1 cells, coincubation with low-dose gentamicin and MitoQ decreased Δψ_m (P = .002). Coincubation of LLC-PK1 cells with high-dose gentamicin and DMSO, MitoQ, or IDB depolarized Δψ_m (P < .0001), with MitoQ depolarizing the Δψ_m to a greater extent than that of IDB (P = .03). In contrast, HEI-OC1 cells demonstrated a hyperpolarized Δψ_m when coincubated with high-dose gentamicin and DMSO, MitoQ, or IDB (P < .001).

Conclusion

The combination of gentamicin and MitoQ holds the potential to disrupt Δψ_m. This suggests a heightened need to monitor for toxicity in patients receiving both agents.

Keywords

aminoglycoside gentamicin ototoxicity mitoquinone antioxidants mitochondrial membrane potential

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