title: Impact of inflammation on chlorpromazine-induced cytotoxicity and cholestatic features in HepaRG cells.
abstract: Several factors are thought to be implicated in the occurrence of idiosyncratic adverse drug reactions. The present work aimed to question as to whether inflammation is a determinant factor in hepatic lesions induced by chlorpromazine (CPZ) using the human HepaRG cell line. An inflammation state was induced by a 24-hour exposure to proinflammatory cytokines interleukin-6 (IL-6) and IL-1β; then the cells were simultaneously treated with CPZ and/or cytokine for 24 hours or daily for 5 days. The inflammatory response was assessed by induction of C-reactive protein and IL-8 transcripts and proteins as well as inhibition of CPZ metabolism and down-regulation of cytochrome 3A4 (CYP3A4) and CYP1A2 transcripts, two major cytochrome P450 (P450) enzymes involved in its metabolism. Most effects of cotreatments with cytokines and CPZ were amplified or only observed after five daily treatments; they mainly included increased cytotoxicity and overexpression of oxidative stress-related genes, decreased Na(+)-taurocholate cotransporting polypeptide mRNA levels and activity, a key transporter involved in bile acids uptake, and deregulation of several other transporters. However, CPZ-induced inhibition of taurocholic acid efflux and pericanalicular F-actin distribution were not affected. In addition, a time-dependent induction of phospholipidosis was noticed in CPZ-treated cells, without obvious influence of the inflammatory stress. In summary, our results show that an inflammatory state induced by proinflammatory cytokines increased cytotoxicity and enhanced some cholestatic features induced by the idiosyncratic drug CPZ in HepaRG cells. These changes, together with inhibition of P450 activities, could have important consequences if extrapolated to the in vivo situation.
authors.affiliation: Inserm UMR991, Foie, Métabolismes et Cancer, Rennes, France (P.B-E.A., A.S., S.A., H.A-A., C.C.S., I.M., C.G-G., A.G.); Université de Rennes 1, Rennes, France (P.B-E.A., A.S., S.A., H.A-A., C.C.S., I.M., C.G-G., A.G.); EDST-PRASE and EDST-AZM-Center-LBA3B, Université Libanaise, Lebanon (P.B-E.A., H.A-A., Z.A-R.); Laboratoire d'Urgence et de Réanimation, Hôpital Pontchaillou, Rennes, France (S.L., I.M.); Sanofi R&D, Alfortville, France (G.L.).
authors.forename: Pamela
authors.initials: P
authors.lastname: Bachour-El Azzi
authors.affiliation: Inserm UMR991, Foie, Métabolismes et Cancer, Rennes, France (P.B-E.A., A.S., S.A., H.A-A., C.C.S., I.M., C.G-G., A.G.); Université de Rennes 1, Rennes, France (P.B-E.A., A.S., S.A., H.A-A., C.C.S., I.M., C.G-G., A.G.); EDST-PRASE and EDST-AZM-Center-LBA3B, Université Libanaise, Lebanon (P.B-E.A., H.A-A., Z.A-R.); Laboratoire d'Urgence et de Réanimation, Hôpital Pontchaillou, Rennes, France (S.L., I.M.); Sanofi R&D, Alfortville, France (G.L.).
authors.forename: Ahmad
authors.initials: A
authors.lastname: Sharanek
authors.affiliation: Inserm UMR991, Foie, Métabolismes et Cancer, Rennes, France (P.B-E.A., A.S., S.A., H.A-A., C.C.S., I.M., C.G-G., A.G.); Université de Rennes 1, Rennes, France (P.B-E.A., A.S., S.A., H.A-A., C.C.S., I.M., C.G-G., A.G.); EDST-PRASE and EDST-AZM-Center-LBA3B, Université Libanaise, Lebanon (P.B-E.A., H.A-A., Z.A-R.); Laboratoire d'Urgence et de Réanimation, Hôpital Pontchaillou, Rennes, France (S.L., I.M.); Sanofi R&D, Alfortville, France (G.L.).
authors.forename: Ziad
authors.initials: Z
authors.lastname: Abdel-Razzak
authors.affiliation: Inserm UMR991, Foie, Métabolismes et Cancer, Rennes, France (P.B-E.A., A.S., S.A., H.A-A., C.C.S., I.M., C.G-G., A.G.); Université de Rennes 1, Rennes, France (P.B-E.A., A.S., S.A., H.A-A., C.C.S., I.M., C.G-G., A.G.); EDST-PRASE and EDST-AZM-Center-LBA3B, Université Libanaise, Lebanon (P.B-E.A., H.A-A., Z.A-R.); Laboratoire d'Urgence et de Réanimation, Hôpital Pontchaillou, Rennes, France (S.L., I.M.); Sanofi R&D, Alfortville, France (G.L.).
authors.forename: Sebastien
authors.initials: S
authors.lastname: Antherieu
authors.affiliation: Inserm UMR991, Foie, Métabolismes et Cancer, Rennes, France (P.B-E.A., A.S., S.A., H.A-A., C.C.S., I.M., C.G-G., A.G.); Université de Rennes 1, Rennes, France (P.B-E.A., A.S., S.A., H.A-A., C.C.S., I.M., C.G-G., A.G.); EDST-PRASE and EDST-AZM-Center-LBA3B, Université Libanaise, Lebanon (P.B-E.A., H.A-A., Z.A-R.); Laboratoire d'Urgence et de Réanimation, Hôpital Pontchaillou, Rennes, France (S.L., I.M.); Sanofi R&D, Alfortville, France (G.L.).
authors.forename: Houssein
authors.initials: H
authors.lastname: Al-Attrache
authors.affiliation: Inserm UMR991, Foie, Métabolismes et Cancer, Rennes, France (P.B-E.A., A.S., S.A., H.A-A., C.C.S., I.M., C.G-G., A.G.); Université de Rennes 1, Rennes, France (P.B-E.A., A.S., S.A., H.A-A., C.C.S., I.M., C.G-G., A.G.); EDST-PRASE and EDST-AZM-Center-LBA3B, Université Libanaise, Lebanon (P.B-E.A., H.A-A., Z.A-R.); Laboratoire d'Urgence et de Réanimation, Hôpital Pontchaillou, Rennes, France (S.L., I.M.); Sanofi R&D, Alfortville, France (G.L.).
authors.forename: Camille C
authors.initials: CC
authors.lastname: Savary
authors.affiliation: Inserm UMR991, Foie, Métabolismes et Cancer, Rennes, France (P.B-E.A., A.S., S.A., H.A-A., C.C.S., I.M., C.G-G., A.G.); Université de Rennes 1, Rennes, France (P.B-E.A., A.S., S.A., H.A-A., C.C.S., I.M., C.G-G., A.G.); EDST-PRASE and EDST-AZM-Center-LBA3B, Université Libanaise, Lebanon (P.B-E.A., H.A-A., Z.A-R.); Laboratoire d'Urgence et de Réanimation, Hôpital Pontchaillou, Rennes, France (S.L., I.M.); Sanofi R&D, Alfortville, France (G.L.).
authors.forename: Sylvie
authors.initials: S
authors.lastname: Lepage
authors.affiliation: Inserm UMR991, Foie, Métabolismes et Cancer, Rennes, France (P.B-E.A., A.S., S.A., H.A-A., C.C.S., I.M., C.G-G., A.G.); Université de Rennes 1, Rennes, France (P.B-E.A., A.S., S.A., H.A-A., C.C.S., I.M., C.G-G., A.G.); EDST-PRASE and EDST-AZM-Center-LBA3B, Université Libanaise, Lebanon (P.B-E.A., H.A-A., Z.A-R.); Laboratoire d'Urgence et de Réanimation, Hôpital Pontchaillou, Rennes, France (S.L., I.M.); Sanofi R&D, Alfortville, France (G.L.).
authors.forename: Isabelle
authors.initials: I
authors.lastname: Morel
authors.affiliation: Inserm UMR991, Foie, Métabolismes et Cancer, Rennes, France (P.B-E.A., A.S., S.A., H.A-A., C.C.S., I.M., C.G-G., A.G.); Université de Rennes 1, Rennes, France (P.B-E.A., A.S., S.A., H.A-A., C.C.S., I.M., C.G-G., A.G.); EDST-PRASE and EDST-AZM-Center-LBA3B, Université Libanaise, Lebanon (P.B-E.A., H.A-A., Z.A-R.); Laboratoire d'Urgence et de Réanimation, Hôpital Pontchaillou, Rennes, France (S.L., I.M.); Sanofi R&D, Alfortville, France (G.L.).
authors.forename: Gilles
authors.initials: G
authors.lastname: Labbe
authors.affiliation: Inserm UMR991, Foie, Métabolismes et Cancer, Rennes, France (P.B-E.A., A.S., S.A., H.A-A., C.C.S., I.M., C.G-G., A.G.); Université de Rennes 1, Rennes, France (P.B-E.A., A.S., S.A., H.A-A., C.C.S., I.M., C.G-G., A.G.); EDST-PRASE and EDST-AZM-Center-LBA3B, Université Libanaise, Lebanon (P.B-E.A., H.A-A., Z.A-R.); Laboratoire d'Urgence et de Réanimation, Hôpital Pontchaillou, Rennes, France (S.L., I.M.); Sanofi R&D, Alfortville, France (G.L.).
authors.forename: Christiane
authors.initials: C
authors.lastname: Guguen-Guillouzo
authors.affiliation: Inserm UMR991, Foie, Métabolismes et Cancer, Rennes, France (P.B-E.A., A.S., S.A., H.A-A., C.C.S., I.M., C.G-G., A.G.); Université de Rennes 1, Rennes, France (P.B-E.A., A.S., S.A., H.A-A., C.C.S., I.M., C.G-G., A.G.); EDST-PRASE and EDST-AZM-Center-LBA3B, Université Libanaise, Lebanon (P.B-E.A., H.A-A., Z.A-R.); Laboratoire d'Urgence et de Réanimation, Hôpital Pontchaillou, Rennes, France (S.L., I.M.); Sanofi R&D, Alfortville, France (G.L.) andre.guillouzo@univ-rennes1.fr.
authors.forename: André
authors.initials: A
authors.lastname: Guillouzo
mesh_terms: D000199:Actins / Q000235:genetics / Q000378:metabolism; D001647:Bile Acids and Salts / Q000235:genetics / Q000378:metabolism; D002097:C-Reactive Protein / Q000235:genetics / Q000378:metabolism; D002460:Cell Line; D002746:Chlorpromazine / Q000009:adverse effects*; D002779:Cholestasis / Q000139:chemically induced / Q000235:genetics / Q000378:metabolism*; D019388:Cytochrome P-450 CYP1A2 / Q000235:genetics / Q000378:metabolism; D051544:Cytochrome P-450 CYP3A / Q000235:genetics / Q000378:metabolism; D015536:Down-Regulation / Q000235:genetics; D006801:Humans; D007249:Inflammation / Q000235:genetics / Q000378:metabolism*; D007378:Interleukins / Q000235:genetics / Q000378:metabolism; D029363:Organic Anion Transporters, Sodium-Dependent / Q000235:genetics / Q000378:metabolism; D018384:Oxidative Stress / Q000235:genetics; D012333:RNA, Messenger / Q000235:genetics; D027981:Symporters / Q000235:genetics / Q000378:metabolism; D013656:Taurocholic Acid / Q000235:genetics / Q000378:metabolism
chemical_list: D000199:Actins; D001647:Bile Acids and Salts; D007378:Interleukins; D029363:Organic Anion Transporters, Sodium-Dependent; D012333:RNA, Messenger; D027981:Symporters; C071908:sodium-bile acid cotransporter; D013656:Taurocholic Acid; D002097:C-Reactive Protein; C504573:CYP1A2 protein, human; D019388:Cytochrome P-450 CYP1A2; D051544:Cytochrome P-450 CYP3A; C510163:CYP3A4 protein, human; D002746:Chlorpromazine