Assessment of mitochondrial membrane potential in HEI-OC1 and LLC-PK1 cells treated with gentamicin and mitoquinone

Otolaryngol Head Neck Surg. 2015 Apr;152(4):729-33. doi: 10.1177/0194599814564934. Epub 2014 Dec 30.

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; antioxidants; gentamicin; mitochondrial membrane potential; mitoquinone; ototoxicity.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antioxidants / pharmacology*
  • Apoptosis / drug effects
  • Cells, Cultured
  • Gentamicins / pharmacology*
  • Humans
  • LLC-PK1 Cells
  • Membrane Potential, Mitochondrial / drug effects*
  • Organophosphorus Compounds / pharmacology*
  • Oxidative Stress / drug effects
  • Prospective Studies
  • Swine
  • Ubiquinone / analogs & derivatives*
  • Ubiquinone / pharmacology

Substances

  • Antioxidants
  • Gentamicins
  • Organophosphorus Compounds
  • Ubiquinone
  • mitoquinone