The effects of intestinal LPS exposure on inflammatory responses in a porcine enterohepatic co-culture system.
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.
Department of Pharmacology and Toxicology, Faculty of Veterinary Science, Szent István University, István u. 2, Budapest, 1078, Hungary, Gere.Erzsebet@aotk.szie.hu.
Erzsebet
E
Paszti-Gere
Gabor
G
Matis
Orsolya
O
Farkas
Anna
A
Kulcsar
Orsolya
O
Palocz
Gyorgy
G
Csiko
Zsuzsanna
Z
Neogrady
Peter
P
Galfi
D000418:Albumins / Q000096:biosynthesis; D000818:Animals; D002460:Cell Line; D002470:Cell Survival; D018920:Coculture Techniques; D019363:Cytochrome P-450 CYP1A1 / Q000096:biosynthesis; D019388:Cytochrome P-450 CYP1A2 / Q000096:biosynthesis; D017097:Electric Impedance; D004847:Epithelial Cells / Q000276:immunology*; D015870:Gene Expression; D020869:Gene Expression Profiling; D022781:Hepatocytes / Q000276:immunology*; D007249:Inflammation / Q000139:chemically induced / Q000276:immunology*; D016209:Interleukin-8 / Q000096:biosynthesis; D007413:Intestinal Mucosa / Q000166:cytology / Q000276:immunology*; D008070:Lipopolysaccharides; D013552:Swine; D014409:Tumor Necrosis Factor-alpha / Q000096:biosynthesis
D000418:Albumins; D016209:Interleukin-8; D008070:Lipopolysaccharides; D014409:Tumor Necrosis Factor-alpha; D019363:Cytochrome P-450 CYP1A1; D019388:Cytochrome P-450 CYP1A2