MilliporeSigma
  • Home
  • Search Results
  • Mitochondrial (dys)function - a factor underlying the variability of efavirenz-induced hepatotoxicity?

Mitochondrial (dys)function - a factor underlying the variability of efavirenz-induced hepatotoxicity?

British journal of pharmacology (2014-11-21)
M Polo, F Alegre, H A Funes, A Blas-Garcia, V M Victor, J V Esplugues, N Apostolova
ABSTRACT

The non-nucleoside analogue reverse transcriptase inhibitor efavirenz is associated with hepatic toxicity and metabolic disturbances. Although the mechanisms involved are not clear, recent evidence has pinpointed a specific mitochondrial action of efavirenz accompanied by the induction of an endoplasmic reticulum (ER) stress/unfolded protein response in human hepatic cells. The aim of this study was to further investigate the involvement of this organelle by evaluating efavirenz's effects in cells lacking functional mitochondria (rho°) and comparing them with those of the typical mitotoxic agent rotenone, a standard complex I inhibitor, and the ER stress inducer thapsigargin. Hep3B rho(+) and rho° cells were treated with clinically relevant concentrations of efavirenz, then mitochondrial function and cytotoxicity were studied using standard cell biology techniques. Efavirenz-treated rho° cells exhibited a substantial reduction in parameters indicative of mitochondrial interference, such as increased superoxide production, mitochondrial mass/morphology alterations and enhanced expression of LONP, a highly conserved mitochondrial protease. In line with these results, the cytotoxic effect (cell number, chromatin condensation, cell cycle alterations and induction of apoptosis) of efavirenz was less pronounced in Hep3B respiration-depleted cells than in wild-type cells. The effect of efavirenz was both similar and different from those of two distinct mitochondrial stressors, thapsigargin and rotenone. Cells lacking normal mitochondria (rho°) are less vulnerable to efavirenz. Our results provide further evidence that the hepatic damage induced by efavirenz involves acute interference with mitochondria and extend our knowledge of the response of mitochondria/ER to a stress stimulus.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Efavirenz, ≥98% (HPLC)
Hydrocortisone for peak identification, European Pharmacopoeia (EP) Reference Standard
Supelco
L-Glutamine, Pharmaceutical Secondary Standard; Certified Reference Material
Sigma-Aldrich
L-Glutamine
Supelco
L-Glutamine, certified reference material, TraceCERT®
Sigma-Aldrich
Ethidium bromide, BioReagent, for molecular biology, powder
Sigma-Aldrich
L-Glutamine, meets USP testing specifications, suitable for cell culture, 99.0-101.0%, from non-animal source
Sigma-Aldrich
Rotenone, ≥95%
Supelco
Rotenone, PESTANAL®, analytical standard
Sigma-Aldrich
L-Glutamine, BioUltra, ≥99.5% (NT)
Sigma-Aldrich
L-Glutamine, γ-irradiated, BioXtra, suitable for cell culture
Sigma-Aldrich
L-Glutamine, ReagentPlus®, ≥99% (HPLC)
Sigma-Aldrich
Thapsigargin, ≥98% (HPLC), solid film
Sigma-Aldrich
Tetramethylrhodamine methyl ester perchlorate, ≥95%
Sigma-Aldrich
Ethidium bromide, ~95% (HPLC)
SAFC
L-Glutamine
Sigma-Aldrich
Propidium iodide solution, solution (1.0 mg/ml in water)
Supelco
Hydrocortisone, Pharmaceutical Secondary Standard; Certified Reference Material
Supelco
Hydrocortisone, VETRANAL®, analytical standard
Sigma-Aldrich
Ethidium bromide solution, for fluorescence, ~1% in H2O
Sigma-Aldrich
Propidium iodide, ≥94% (HPLC)
Sigma-Aldrich
Uridine, powder, BioReagent, suitable for cell culture
Sigma-Aldrich
Uridine, BioUltra, ≥99%
Sigma-Aldrich
Hydrocortisone, γ-irradiated, powder, BioXtra, suitable for cell culture
Sigma-Aldrich
Hydrocortisone, ≥98% (HPLC)
Sigma-Aldrich
Hydrocortisone, meets USP testing specifications
Sigma-Aldrich
Hydrocortisone, BioReagent, suitable for cell culture
Sigma-Aldrich
Propidium iodide, ≥94.0% (HPLC)
Sigma-Aldrich
Uridine, ≥99%
Sigma-Aldrich
Ethidium bromide solution, BioReagent, for molecular biology, 500 μg/mL in H2O