Inhibition of mitochondrial function by efavirenz increases lipid content in hepatic cells.
Efavirenz (EFV) is a non-nucleoside reverse transcriptase inhibitor (NNRTI) widely used in human immunodeficiency virus (HIV) infection therapy. It has been associated with hepatotoxic effects and alterations in lipid and body fat composition. Given the importance of the liver in lipid regulation, we have evaluated the effects of clinically used concentrations of EFV on the mitochondria and lipid metabolism of human hepatic cells in vitro. Mitochondrial function was rapidly undermined by EFV to an extent that varied with the concentration employed; in particular, respiration and intracellular adenosine triphosphate (ATP) levels were reduced whereas reactive oxygen species (ROS) production increased. Results in isolated mitochondria suggest that the mechanism responsible for these actions was a specific inhibition of complex I of the respiratory chain. The reduction in energy production triggered a compensatory mechanism mediated by the enzyme adenosine monophosphate-activated protein kinase (AMPK), the master switch of cellular bioenergetics. Fluorescence and nuclear magnetic resonance demonstrated a rapid intracellular increase of neutral lipids, usually in the form of droplets. This was prevented by the AMPK inhibitor compound C and by removal of fatty acids from the culture medium. These effects were not reproduced by Nevirapine, another NNRTI. EFV is clinically coadministered with two nucleoside reverse transcriptase inhibitors. Evaluation of one of the most common combination, EFV/Lamivudine/Abacavir, revealed that the effects of EFV on ROS production were enhanced.


CONCLUSIONS
Clinical concentrations of EFV induce bioenergetic stress in hepatic cells by acutely inhibiting mitochondrial function. This new mechanism of mitochondrial interference leads to an accumulation of lipids in the cytoplasm that is mediated by activation of AMPK.
Blas-García
Ana
A
Departamento de Farmacología, Facultad de Medicina, Universidad de Valencia-CIBERehd, Valencia, Spain.
Apostolova
Nadezda
N
Ballesteros
Daniel
D
Monleón
Daniel
D
Morales
Jose M
JM
Rocha
Milagros
M
Victor
Victor M
VM
Esplugues
Juan V
JV
D000091162:AMP-Activated Protein Kinase Kinases; D000480:Alkynes; D000818:Animals; D019380:Anti-HIV Agents / Q000494:pharmacology*; D048588:Benzoxazines / Q000494:pharmacology*; D003521:Cyclopropanes; D022781:Hepatocytes / Q000187:drug effects* / Q000378:metabolism; D006801:Humans; D050356:Lipid Metabolism / Q000187:drug effects*; D008055:Lipids / Q000032:analysis; D008297:Male; D008930:Mitochondria, Liver / Q000187:drug effects* / Q000502:physiology; D010101:Oxygen Consumption / Q000187:drug effects; D011494:Protein Kinases / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley
D000480:Alkynes; D019380:Anti-HIV Agents; D048588:Benzoxazines; D003521:Cyclopropanes; D008055:Lipids; D011494:Protein Kinases; D000091162:AMP-Activated Protein Kinase Kinases; C098320:efavirenz