title: Characterization of chemical-induced sterile inflammation in vitro: application of the model compound ketoconazole in a human hepatic co-culture system.
abstract: Liver injury as a result of a sterile inflammation is closely linked to the activation of immune cells, including macrophages, by damaged hepatocytes. This interaction between immune cells and hepatocytes is as yet not considered in any of the in vitro test systems applied during the generation of new drugs. Here, we established and characterized a novel in vitro co-culture model with two human cell lines, HepG2 and differentiated THP-1. Ketoconazole, an antifungal drug known for its hepatotoxicity, was used as a model compound in the testing of the co-culture. Single cultures of HepG2 and THP-1 cells were studied as controls. Different metabolism patterns of ketoconazole were observed for the single and co-culture incubations as well as for the different cell types. The main metabolite N-deacetyl ketoconazole was found in cell pellets, but not in supernatants of cell cultures. Global proteome analysis showed that the NRF2-mediated stress response and the CXCL8 (IL-8) pathway were induced by ketoconazole treatment under co-culture conditions. The upregulation and ketoconazole-induced secretion of several pro-inflammatory cytokines, including CXCL8, TNF-α and CCL3, was observed in the co-culture system only, but not in single cell cultures. Taking together, we provide evidence that the co-culture model applied might be suitable to serve as tool for the prediction of chemical-induced sterile inflammation in liver tissue in vivo.
authors.affiliation: Department of Chemical and Product Safety, German Federal Institute for Risk Assessment (BfR), Max-Dohrn-Strasse 8-10, 10589, Berlin, Germany. franziska.wewering@bfr.bund.de.
authors.forename: Franziska
authors.initials: F
authors.lastname: Wewering
authors.affiliation: Department of Proteomics, UFZ, Helmholtz-Centre for Environmental Research, Permoserstrasse 15, 04318, Leipzig, Germany.
authors.forename: Florent
authors.initials: F
authors.lastname: Jouy
authors.affiliation: Department of Metabolomics, UFZ, Helmholtz-Centre for Environmental Research, Permoserstrasse 15, 04318, Leipzig, Germany.|Institute of Forensic Medicine, Jena University Hospital, 07743, Jena, Germany.
authors.forename: Dirk K
authors.initials: DK
authors.lastname: Wissenbach
authors.affiliation: Department of Metabolomics, UFZ, Helmholtz-Centre for Environmental Research, Permoserstrasse 15, 04318, Leipzig, Germany.
authors.forename: Scarlett
authors.initials: S
authors.lastname: Gebauer
authors.affiliation: Clinic for Endocrinology and Nephrology, Faculty of Medicine, University of Leipzig, Liebigstrasse 20, 04103, Leipzig, Germany.
authors.forename: Matthias
authors.initials: M
authors.lastname: Blüher
authors.affiliation: Institute of Biochemistry, Faculty of Medicine, University of Leipzig, Johannisallee 30, 04103, Leipzig, Germany.
authors.forename: Rolf
authors.initials: R
authors.lastname: Gebhardt
authors.affiliation: Department of Chemical and Product Safety, German Federal Institute for Risk Assessment (BfR), Max-Dohrn-Strasse 8-10, 10589, Berlin, Germany.
authors.forename: Ralph
authors.initials: R
authors.lastname: Pirow
authors.affiliation: Department of Proteomics, UFZ, Helmholtz-Centre for Environmental Research, Permoserstrasse 15, 04318, Leipzig, Germany.|Department of Metabolomics, UFZ, Helmholtz-Centre for Environmental Research, Permoserstrasse 15, 04318, Leipzig, Germany.|Department of Life Sciences and Chemistry, Centre for Microbial Communities, University of Aalborg, Sohngaardsholmsvej 49, 9000, Aalborg, Denmark.|Faculty of Biosciences, Pharmacy and Psychology, Institute of Biochemistry, University of Leipzig, Brüderstrasse 34, 04103, Leipzig, Germany.
authors.forename: Martin
authors.initials: M
authors.lastname: von Bergen
authors.affiliation: Department of Proteomics, UFZ, Helmholtz-Centre for Environmental Research, Permoserstrasse 15, 04318, Leipzig, Germany.|Department of Bioanalytics, University of Applied Sciences and Arts of Coburg, 96450, Coburg, Germany.
authors.forename: Stefan
authors.initials: S
authors.lastname: Kalkhof
authors.affiliation: Department of Chemical and Product Safety, German Federal Institute for Risk Assessment (BfR), Max-Dohrn-Strasse 8-10, 10589, Berlin, Germany.
authors.forename: Andreas
authors.initials: A
authors.lastname: Luch
authors.affiliation: Department of Chemical and Product Safety, German Federal Institute for Risk Assessment (BfR), Max-Dohrn-Strasse 8-10, 10589, Berlin, Germany.
authors.forename: Sebastian
authors.initials: S
authors.lastname: Zellmer
mesh_terms: D056486:Chemical and Drug Induced Liver Injury / Q000378:metabolism / Q000473:pathology*; D018920:Coculture Techniques; D056945:Hep G2 Cells / Q000187:drug effects; D006801:Humans; D016209:Interleukin-8 / Q000378:metabolism; D007654:Ketoconazole / Q000009:adverse effects* / Q000031:analogs & derivatives / Q000378:metabolism / Q000493:pharmacokinetics; D053078:Membrane Potential, Mitochondrial / Q000187:drug effects; D016328:NF-kappa B / Q000378:metabolism; D011506:Proteins / Q000032:analysis / Q000378:metabolism; D017382:Reactive Oxygen Species / Q000378:metabolism; D018675:Toxicity Tests / Q000379:methods*; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism
chemical_list: C508606:CXCL8 protein, human; D016209:Interleukin-8; D016328:NF-kappa B; D011506:Proteins; D017382:Reactive Oxygen Species; D014409:Tumor Necrosis Factor-alpha; C035996:deacylketoconazole; D007654:Ketoconazole
keywords: CXCL8; HepG2; Metabolism; NFκB; NRF2; ROS; THP-1