Drug-induced hepatic steatosis in absence of severe mitochondrial dysfunction in HepaRG cells: proof of multiple mechanism-based toxicity

Julien Allard (1) , Simon Bucher (1) , Julie Massart (1) , Pierre-Jean Ferron (1) , Dounia Le Guillou (1) , Roxane Loyant (1, 2) , Yoann Daniel (1) , Youenn Launay (1) , Nelly Buron (1, 2) , Karima Begriche (1) , Annie Borgne-Sanchez (1, 2) , Bernard Fromenty (1)

1 NuMeCan - Nutrition, Métabolismes et Cancer
2 Mitologics S.A.S.

Steatosis is a liver lesion reported with numerous pharmaceuticals. Prior studies showed that severe impairment of mitochondrial fatty acid oxidation (mtFAO) constantly leads to lipid accretion in liver. However, much less is known about the mechanism(s) of drug-induced steatosis in the absence of severe mitochondrial dysfunction, although previous studies suggested the involvement of mild-to-moderate inhibition of mtFAO, increased de novo lipogenesis (DNL), and impairment of very low-density lipoprotein (VLDL) secretion. The objective of our study, mainly carried out in human hepatoma HepaRG cells, was to investigate these 3 mechanisms with 12 drugs able to induce steatosis in human amiodarone (AMIO, used as positive control), allopurinol (ALLO),d-penicillamine (DPEN), 5-fluorouracil (5FU), indinavir (INDI), indomethacin (INDO), methimazole (METHI), methotrexate (METHO), nifedipine (NIF), rifampicin (RIF), sulindac (SUL), and troglitazone (TRO). Hepatic cells were exposed to drugs for 4 days with concentrations decreasing ATP level by less than 30% as compared to control and not exceeding 100 x C-max. Among the 12 drugs, AMIO, ALLO, 5FU, INDI, INDO, METHO, RIF, SUL, and TRO induced steatosis in HepaRG cells. AMIO, INDO, and RIF decreased mtFAO. AMIO, INDO, and SUL enhanced DNL. ALLO, 5FU, INDI, INDO, SUL, RIF, and TRO impaired VLDL secretion. These seven drugs reduced the mRNA level of genes playing a major role in VLDL assembly and also induced endoplasmic reticulum (ER) stress. Thus, in the absence of severe mitochondrial dysfunction, drug-induced steatosis can be triggered by different mechanisms, although impairment of VLDL secretion seems more frequently involved, possibly as a consequence of ER stress.

Mots clés: Very low-density lipoprotein; Endoplasmic reticulum stress; Lipogenesis; Fatty acid oxidation; Steatosis; Mitochondria

Domaines: Pharmacologie; Hépatologie et Gastroentérologie

Julien Allard, Simon Bucher, Julie Massart, Pierre-Jean Ferron, Dounia Le Guillou, et al.. Drug-induced hepatic steatosis in absence of severe mitochondrial dysfunction in HepaRG cells: proof of multiple mechanism-based toxicity. Cell Biology and Toxicology, 2021, 37 (2), pp.151-175. ⟨10.1007/s10565-020-09537-1⟩. ⟨hal-02886702⟩