Differential sensitivity of metabolically competent and non-competent HepaRG cells to apoptosis induced by diclofenac combined or not with TNF-α. (6th September 2016)

• Record Type: Journal Article
• Authors: Al-Attrache, Houssein; Sharanek, Ahmad; Burban, Audrey; Burbank, Matthew; Gicquel, Thomas; Abdel-Razzak, Ziad; Guguen-Guillouzo, Christiane; Morel, Isabelle; Guillouzo, André
• Is Part Of: Toxicology letters. Volume 258(2016)
• Page Start: 71
• Page End: 86
• Publication Date: 2016-09-06
• Subjects: ABT 1-aminobenzotriazole -- Ac-DEVD-AMC N-acetyl-Asp-Glu-Val-Asp-7-amido-4-methylcoumarin -- AC-IETD-AMC Ac-Ile-Glu-Thr-Asp-7-amino-4-methylcoumarin -- AC-LEHD-AMC Ac-Leu-Glu-His-Asp-7-Amino-4-methylcoumarin -- BSEP bile salt export pump -- CDF 5 (and 6)-carboxy-2′, 7′-dichlorofluorescein (CDF) -- CRP C-reactive protein -- CYP cytochrome P450 -- DCF diclofenac -- DMSO dimethyl sulfoxide -- ER endoplasmic reticulum -- H2-DCFDA 2′, 7′-dichlorodihydrofluorescein -- HO1 heme oxygenase 1 -- IL-8 interleukin-8 -- MnSOD manganese superoxide dismutase -- MRP2 multidrug associated protein 2 -- MTT methylthiazoltetrazolium -- NAC N-acetyl cysteine -- NTCP Na+-dependent taurocholate cotransporting polypeptide -- PBS phosphate buffered saline -- ROS reactive oxygen species -- RT-qPCR real-time quantitative polymerase chain reaction -- TNF-α tumor necrosis factor α; Drug metabolism -- Differentiation status -- Tumor necrosis factor α -- Caspases -- Reactive oxygen species -- Endoplasmic reticulum stress -- Cholestasis -- Glutathione transferases -- HepG2 cells -- Primary human hepatocytes; Toxicology -- Periodicals; 363.179
• DOI: 10.1016/j.toxlet.2016.06.008
• Languages: English
• ISSNs: 0378-4274

Abstract

Highlights: HepaRG hepatocytes are less sensitive to DCF than HepG2 cells and undifferentiated HepaRG cells. The lower sensitivity of HepaRG hepatocytes is related to their high detoxifying capacity. Inhibition of glutathione transferases results in increased DCF cytotoxicity. TNF-α potentiation of DCF cytotoxicity is not observed in undifferentiated HepaRG cells. DCF-induced cholestasis typified by bile canaliculi dilatation is not aggravated by TNF-α. Abstract: 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 24 h-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.