A porcine enterohepatic co-culture system, with primary hepatocytes as bottom layer and IPEC-J2 epithelial cells as upper layer, was developed to study the effects of lipopolysaccharides (LPS) on the gene expression profile of pro-inflammatory cytokines (interleukin-8 (IL-8) and tumor necrosis factor-α) and CYP enzymes (CYP1A1, CYP1A2, CYP3A29). The barrier integrity of IPEC-J2 cells was investigated by transepithelial electrical resistance measurements and by fluorescein isothiocyanate–dextran-based test. Basolateral IL-8 production was significantly elevated in LPS-treated IPEC-J2 and primary hepatocyte mono-cultures as well as in the co-culture system, in a dose-independent manner. The LPS-induced changes in the expression of the CYP1A2 and CYP3A29 genes in hepatocyte mono-cultures differed from those in co-culture after LPS treatment on the apical side of the IPEC-J2 cell layer. CYP1A2 was downregulated by the LPS treatment in mono-cultures but upregulated at 10 μg/ml LPS in co-culture; gene expression of CYP3A29 showed no significant LPS-induced change in the hepatocyte mono-culture but was significantly downregulated in co-culture. The newly established co-culture system capable of mimicking enterohepatic interplay in LPS-induced inflammatory responses in vitro can be used in the future for reliable screening of potential anti-inflammatory compounds.
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The research described here was supported by the Hungarian Scientific Research Fund (OTKA grants no. 100701 and no. 105718). The study was sponsored by the TÁMOP-4.2.2.B-10/1 and TÁMOP-4.2.1.B-11/2/KMR-2011-0003 projects. This paper is dedicated to Prof. Dr. Péter Gálfi on the occasion of his 60th birthday for establishment of the IPEC-J2 and hepatocyte co-culture system. We are indebted to Dr. Jody Gookin (Department of Clinical Sciences, College of Veterinary Medicine, North Carolina State University, Raleigh, NC, USA) for providing the IPEC-J2 cells. Special thanks have to be given to Hedvig Fébel (Research Institute for Animal Breeding and Nutrition, Herceghalom, Hungary) for providing the animals used for hepatocyte isolation. The excellent assistance of Janka Petrilla and Melinda Szabó (Faculty of Veterinary Science, Szent István University, Budapest, Hungary) is gratefully acknowledged.
Erzsebet Paszti-Gere and Gabor Matis have equal contribution to the manuscript.
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Paszti-Gere, E., Matis, G., Farkas, O. et al. The Effects of Intestinal LPS Exposure on Inflammatory Responses in a Porcine Enterohepatic Co-culture System. Inflammation 37, 247–260 (2014). https://doi.org/10.1007/s10753-013-9735-7
- enterocyte-liver co-culture
- lipopolysaccharide (LPS)
- inflammatory cytokines
- cytochrome P450 (CYP) enzymes