Differential sensitivity of metabolically competent and non-competent HepaRG cells to apoptosis induced by diclofenac combined or not with TNF-α.
The role of reactive metabolites and inflammatory stress has been largely evoked in idiosyncratic hepatotoxicity of diclofenac (DCF); however mechanisms remain poorly understood. We aimed to evaluate the influence of liver cell phenotype on the hepatotoxicity of DCF combined or not with TNF-α using differentiated and undifferentiated HepaRG cells, and for comparison, HepG2 cells. Our results demonstrate that after a 24h-treatment metabolizing HepaRG cells were less sensitive to DCF than their undifferentiated non-metabolizing counterparts as shown by lower oxidative and endoplasmic reticulum stress responses and lower activation of caspase 9. Differentiated HepaRG cells were also less sensitive than HepG2 cells. Their lower sensitivity to DCF was related to their high content in glutathione transferases. DCF-induced apoptotic effects were potentiated by TNF-α only in death receptor-expressing differentiated HepaRG and HepG2 cells and were associated with marked activation of caspase 8. TNF-α co-treatment did not aggravate DCF-induced cholestatic features. Altogether, our results demonstrate that (i) lower sensitivity to DCF of differentiated HepaRG cells compared to their non-metabolically active counterparts was related to their high detoxifying capacity, giving support to the higher sensitivity of nonhepatic tissues than liver to this drug; (ii) TNF-α-potentiation of DCF cytotoxicity occurred only in death receptor-expressing cells.
Inserm U991, Faculty of Pharmacy, Rennes, France; Rennes 1 University, Rennes, France; Laboratory of Applied Biotechnology: biomolecules, biotherapy and bioprocesses (LBA3B), AZM Center-Tripoli and EDST-PRASE-Beirut, Rafic Hariri Campus-Lebanese University, Lebanon.
Houssein
H
Al-Attrache
Inserm U991, Faculty of Pharmacy, Rennes, France; Rennes 1 University, Rennes, France.
Ahmad
A
Sharanek
Inserm U991, Faculty of Pharmacy, Rennes, France; Rennes 1 University, Rennes, France.
Audrey
A
Burban
Inserm U991, Faculty of Pharmacy, Rennes, France; Rennes 1 University, Rennes, France.
Matthew
M
Burbank
Inserm U991, Faculty of Pharmacy, Rennes, France; Rennes 1 University, Rennes, France; Laboratory Emergency and Intensive Care, Hospital Pontchaillou, Rennes, France.
Thomas
T
Gicquel
Laboratory of Applied Biotechnology: biomolecules, biotherapy and bioprocesses (LBA3B), AZM Center-Tripoli and EDST-PRASE-Beirut, Rafic Hariri Campus-Lebanese University, Lebanon.
Ziad
Z
Abdel-Razzak
Inserm U991, Faculty of Pharmacy, Rennes, France; Rennes 1 University, Rennes, France.
Christiane
C
Guguen-Guillouzo
Inserm U991, Faculty of Pharmacy, Rennes, France; Rennes 1 University, Rennes, France; Laboratory Emergency and Intensive Care, Hospital Pontchaillou, Rennes, France.
Isabelle
I
Morel
Inserm U991, Faculty of Pharmacy, Rennes, France; Rennes 1 University, Rennes, France. Electronic address: Andre.Guillouzo@univ-rennes1.fr.
André
A
Guillouzo
D000894:Anti-Inflammatory Agents, Non-Steroidal / Q000819:agonists / Q000378:metabolism / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects*; D001711:Biotransformation / Q000187:drug effects; D002454:Cell Differentiation; D002460:Cell Line; D002478:Cells, Cultured; D004008:Diclofenac / Q000819:agonists / Q000378:metabolism / Q000494:pharmacology*; D004351:Drug Resistance; D059865:Endoplasmic Reticulum Stress / Q000187:drug effects; D004791:Enzyme Inhibitors; D005982:Glutathione Transferase / Q000037:antagonists & inhibitors / Q000378:metabolism; D056945:Hep G2 Cells; D022781:Hepatocytes / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D006801:Humans; D020128:Inhibitory Concentration 50; D007700:Kinetics; D008858:Microscopy, Phase-Contrast; D018384:Oxidative Stress / Q000187:drug effects; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism*; D019014:fas Receptor / Q000819:agonists* / Q000378:metabolism
D000894:Anti-Inflammatory Agents, Non-Steroidal; D004791:Enzyme Inhibitors; D014409:Tumor Necrosis Factor-alpha; D019014:fas Receptor; D004008:Diclofenac; D005982:Glutathione Transferase
Caspases; Cholestasis; Differentiation status; Drug metabolism; Endoplasmic reticulum stress; Glutathione transferases; HepG2 cells; Primary human hepatocytes; Reactive oxygen species; Tumor necrosis factor α