Comparative study of CYP2B1/2 induction and the transport of bilirubin and taurocholate in rat hepatocyte-mono- and hepatocyte-Kupffer cell co-cultures

• Published in: Journal of pharmacological and toxicological methods Vol. 82; pp. 1 - 8
• Main Authors: Bátai-Konczos, Attila, Veres, Zsuzsa, Szabó, Mónika, Ioja, Enikő, László, Glória, Török, György, Homolya, László, Jemnitz, Katalin
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 01.11.2016
• Subjects: Animals; Aryl Hydrocarbon Hydroxylases - biosynthesis; Bilirubin - metabolism; Biological Transport; Chemical and Drug Induced Liver Injury - enzymology; Chemical and Drug Induced Liver Injury - metabolism; Coculture Techniques; CYP2B1/2; Cytochrome P-450 CYP2B1 - biosynthesis; Drug Interactions; Enzyme Induction; Hepatocyte; Hepatocytes - drug effects; Hepatocytes - enzymology; Kupffer cell; Kupffer Cells - drug effects; Kupffer Cells - enzymology; Lipopolysaccharides - pharmacology; Male; Metabolic Detoxication, Phase I; Models, Biological; Rats; Rats, Wistar; Sandwich co-culture method; Steroid Hydroxylases - biosynthesis; Taurocholic Acid - metabolism; Transporters; PBS; B; Bsep; Sandwich co-culture method; Mrp; LPS; TC; Kupffer cell; PB; Hepatocyte; PTX; Ntcp; CYP2B1/2; KC; Transporters; GdCl3; CYP; HBSS; H/KC; HPLC
• ISSN: 1056-8719; 1873-488X
• DOI: 10.1016/j.vascn.2016.05.015

Hepatocyte-Kupffer cell (KC) co-cultures represent a promising approach for in vitro modeling of complex interactions between parenchymal and non-parenchymal cells in the liver, responsible for drug-induced liver injury (DILI). In this study we aimed to compare hepatocyte monocultures with hepatocyte-KC co-cultures regarding some basic liver functions associated with the chemical defense system. These pathways involve transporters and enzymes the function of which is highly sensitive towards hepatotoxic events. CYP2B1/2 induction and the biliary and sinusoidal elimination of bilirubin (B) and taurocholate (TC) were studied in rat hepatocyte sandwich cultures compared with rat hepatocyte-KC sandwich co-cultures of 1:0, 6:1, 2:1 and 1:1 cell combinations representing the physiologic and pathologic conditions of the liver. KCs decreased phenobarbital inducibility of CYP2B1/2 in a cell ratio dependent manner and activation of KCs by lipopolisacharide (LPS) amplified this effect. Similarly, KCs decreased the transport of B and its glucuronides (BG) in both sinusoidal and canalicular directions resulting in its intracellular accumulation. In contrast, the uptake and the efflux of TC were greater in the co-cultures than in the hepatocyte monocultures. Immuno-labelling of sodium-dependent taurocholate transporter (Ntcp) revealed increased expression of the transporter in the presence of KCs. Here we presented that KCs have a direct impact on some hepatocyte functions suggesting that the co-culture model may be more suitable for drug related hepatotoxicity studies than hepatocyte monocultures.