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Inflammation
Published in: Inflammation 1/2014

01-02-2014

The Effects of Intestinal LPS Exposure on Inflammatory Responses in a Porcine Enterohepatic Co-culture System

Authors: Erzsebet Paszti-Gere, Gabor Matis, Orsolya Farkas, Anna Kulcsar, Orsolya Palocz, Gyorgy Csiko, Zsuzsanna Neogrady, Peter Galfi

Published in: Inflammation | Issue 1/2014

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Abstract

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.
Literature
1.
Langerholc, T., P.A. Maragkoudakis, J. Wollgast, L. Gradisnik, and A. Cencic. 2011. Novel and established intestinal cell line models—an indispensable tool in food science and nutrition. Trends in Food Science & Technology 22: S11–S20.CrossRef
2.
Oswald, I.P. 2006. Role of intestinal epithelial cells in the innate immune defence of the pig intestine. Veterinary Research 37: 359–368.PubMedCrossRef
3.
Burkey, T.E., K.A. Skjolaas, S.S. Dritz, and J.E. Minton. 2007. Expression of Toll-like receptors, interleukin 8, macrophage migration inhibitory factor, and osteopontin in tissues from pigs challenged with Salmonella enterica serovar Typhimurium or serovar Choleraesuis. Veterinary Immunology and Immunopathology 115: 309–319.PubMedCrossRef
4.
Didierlaurent, A., J.C. Sirard, J.P. Kraehenbuhl, and M.R. Neutra. 2002. How the gut senses its content. Cellular Microbiology 4: 61–72.PubMedCrossRef
5.
Backhed, F., S. Normark, E.K. Schweda, S. Oscarson, and A. Richter-Dahlfors. 2003. Structural requirements for TLR4-mediated LPS signalling: a biological role for LPS modifications. Microbes and Infection 5: 1057–1063.PubMedCrossRef
6.
Ogle, C.K., X.L. Guo, P.O. Hasselgren, J.D. Ogle, and J.W. Alexander. 1997. The gut as a source of inflammatory cytokines after stimulation with endotoxin. European Journal of Surgery 163: 45–51.
7.
Feuerstein, G.Z., T. Liu, and F.C. Barone. 1994. Cytokines, inflammation, and brain injury: role of tumor necrosis factor-alpha. Cerebrovascular and Brain Metabolism Reviews 6: 341–360.PubMed
8.
Hassoun, H.T., B.C. Kone, D.W. Mercer, F.G. Moody, N.W. Weisbrodt, and F.A. Moore. 2001. Post-injury multiple organ failure: the role of the gut. Shock 15: 1–10.PubMedCrossRef
9.
Berschneider, H.M. 1989. Abstract of the Annual Meeting of the American Gastroenterological Association. Digestive Disease Week and the 90th annual meeting of the American Gastroenterological Association, 774. Washington, DC: Elsevier.
10.
Schierack, P., M. Nordhoff, M. Pollmann, K.D. Weyrauch, S. Amasheh, U. Lodemann, J. Jores, B. Tachu, S. Kleta, A. Blikslager, et al. 2006. Characterization of a porcine intestinal epithelial cell line for in vitro studies of microbial pathogenesis in swine. Histochemistry and Cell Biology 125: 293–305.PubMedCrossRef
11.
Paszti-Gere, E., E. Csibrik-Nemeth, K. Szeker, R. Csizinszky, O. Palocz, O. Farkas, and P. Galfi. 2013. Lactobacillus plantarum 2142 prevents intestinal oxidative stress in optimized in vitro systems. Acta Physiologica Hungarica 100: 89–98.PubMedCrossRef
12.
Arce, C., M. Ramirez-Boo, C. Lucena, and J.J. Garrido. 2010. Innate immune activation of swine intestinal epithelial cell lines (IPEC-J2 and IPI-2I) in response to LPS from Salmonella typhimurium. Comparative Immunology, Microbiology and Infectious Diseases 33: 161–174.PubMedCrossRef
13.
Paszti-Gere, E., E. Csibrik-Nemeth, K. Szeker, R. Csizinszky, C. Jakab, and P. Galfi. 2012. Acute oxidative stress affects IL-8 and TNF-alpha expression in IPEC-J2 porcine epithelial cells. Inflammation 35: 994–1004.PubMedCrossRef
14.
Paszti-Gere, E., K. Szeker, E. Csibrik-Nemeth, R. Csizinszky, A. Marosi, O. Palocz, O. Farkas, and P. Galfi. 2012. Metabolites of Lactobacillus plantarum 2142 prevent oxidative stress-induced overexpression of proinflammatory cytokines in IPEC-J2 cell line. Inflammation 35: 1487–1499.PubMedCrossRef
15.
Angrisano, T., R. Pero, S. Peluso, S. Keller, S. Sacchetti, C.B. Bruni, L. Chiariotti, and F. Lembo. 2010. LPS-induced IL-8 activation in human intestinal epithelial cells is accompanied by specific histone H3 acetylation and methylation changes. BMC Microbiology 10: 172.PubMedPubMedCentralCrossRef
16.
Hanson, P.J., A.P. Moran, and K. Butler. 2011. Paracellular permeability is increased by basal lipopolysaccharide in a primary culture of colonic epithelial cells; an effect prevented by an activator of Toll-like receptor-2. Innate Immunity 17: 269–282.PubMedCrossRef
17.
Yu, L.C., A.N. Flynn, J.R. Turner, and A.G. Buret. 2005. SGLT-1-mediated glucose uptake protects intestinal epithelial cells against LPS-induced apoptosis and barrier defects: a novel cellular rescue mechanism? FASEB Journal 19: 1822–1835.PubMedCrossRef
18.
Morgan, E.T. 1997. Regulation of cytochromes P450 during inflammation and infection. Drug Metabolism Reviews 29: 1129–1188.PubMedCrossRef
19.
Sewer, M.B., D.R. Koop, and E.T. Morgan. 1997. Differential inductive and suppressive effects of endotoxin and particulate irritants on hepatic and renal cytochrome P-450 expression. Journal of Pharmacology and Experimental Therapeutics 280: 1445–1454.PubMed
20.
Morgan, E.T. 2001. Regulation of cytochrome p450 by inflammatory mediators: why and how? Drug Metabolism and Disposition 29: 207–212.PubMed
21.
Haller, D., C. Bode, W.P. Hammes, A.M. Pfeifer, E.J. Schiffrin, and S. Blum. 2000. Non-pathogenic bacteria elicit a differential cytokine response by intestinal epithelial cell/leucocyte co-cultures. Gut 47: 79–87.PubMedPubMedCentralCrossRef
22.
Westendorf, A.M., D. Fleissner, W. Hansen, and J. Buer. 2010. T cells, dendritic cells and epithelial cells in intestinal homeostasis. International Journal of Medical Microbiology 300: 11–18.PubMedCrossRef
23.
Cencic, A., and T. Langerholc. 2010. Functional cell models of the gut and their applications in food microbiology—a review. International Journal of Food Microbiology 141(Suppl 1): S4–14.PubMedPubMedCentralCrossRef
24.
Mahler, G.J., M.L. Shuler, and R.P. Glahn. 2009. Characterization of Caco-2 and HT29-MTX cocultures in an in vitro digestion/cell culture model used to predict iron bioavailability. Journal of Nutritional Biochemistry 20: 494–502.CrossRefPubMed
25.
Heredi-Szabo, K., H. Glavinas, E. Kis, D. Mehn, G. Bathori, Z. Veres, L. Kobori, O. von Richter, K. Jemnitz, and P. Krajcsi. 2009. Multidrug resistance protein 2-mediated estradiol-17beta-D-glucuronide transport potentiation: in vitro–in vivo correlation and species specificity. Drug Metabolism and Disposition 37: 794–801.PubMedCrossRef
26.
Begue, J.M., C. Guguen-Guillouzo, N. Pasdeloup, and A. Guillouzo. 1984. Prolonged maintenance of active cytochrome P-450 in adult rat hepatocytes co-cultured with another liver cell type. Hepatology 4: 839–842.PubMedCrossRef
27.
Harimoto, M., M. Yamato, M. Hirose, C. Takahashi, Y. Isoi, A. Kikuchi, and T. Okano. 2002. Novel approach for achieving double-layered cell sheets co-culture: overlaying endothelial cell sheets onto monolayer hepatocytes utilizing temperature-responsive culture dishes. Journal of Biomedical Materials Research 62: 464–470.PubMedCrossRef
28.
Brand, R.M., T.L. Hannah, C. Mueller, Y. Cetin, and F.G. Hamel. 2000. A novel system to study the impact of epithelial barriers on cellular metabolism. Annals of Biomedical Engineering 28: 1210–1217.PubMedCrossRef
29.
Choi, S.H., M. Nishikawa, A. Sakoda, and Y. Sakai. 2004. Feasibility of a simple double-layered coculture system incorporating metabolic processes of the intestine and liver tissue: application to the analysis of benzo[a]pyrene toxicity. Toxicology in Vitro 18: 393–402.PubMedCrossRef
30.
Castell-Auvi, A., M.J. Motilva, A. Macia, H. Torrell, C. Blade, M. Pinent, L. Arola, and A. Ardevol. 2010. Organotypic co-culture system to study plant extract bioactivity on hepatocytes. Food Chemistry 122: 775–781.CrossRef
31.
Puviani, A.C., C. Ottolenghi, B. Tassinari, P. Pazzi, and E. Morsiani. 1998. An update on high-yield hepatocyte isolation methods and on the potential clinical use of isolated liver cells. Comparative Biochemistry and Physiology Part A: Molecular & Integrative Physiology 121: 99–109.CrossRef
32.
Meng, F.Y., Z.S. Chen, M. Han, X.P. Hu, X.X. He, Y. Liu, W.T. He, W. Huang, H. Guo, and P. Zhou. 2010. Porcine hepatocyte isolation and reversible immortalization mediated by retroviral transfer and site-specific recombination. World Journal of Gastroenterology 16: 1660–1664.PubMedPubMedCentralCrossRef
33.
Repetto, G., A. del Peso, and J.L. Zurita. 2008. Neutral red uptake assay for the estimation of cell viability/cytotoxicity. Nature Protocols 3: 1125–1131.PubMedCrossRef
34.
35.
Seki, E., and B. Schnabl. 2012. Role of innate immunity and the microbiota in liver fibrosis: crosstalk between the liver and gut. Journal of Physiology 590: 447–458.CrossRefPubMed
36.
Ilan, Y. 2012. Leaky gut and the liver: a role for bacterial translocation in nonalcoholic steatohepatitis. World Journal of Gastroenterology 18: 2609–2618.PubMedPubMedCentralCrossRef
37.
Richardson, T.A., M. Sherman, L. Antonovic, S.S. Kardar, H.W. Strobel, D. Kalman, and E.T. Morgan. 2006. Hepatic and renal cytochrome p450 gene regulation during Citrobacter rodentium infection in wild-type and toll-like receptor 4 mutant mice. Drug Metabolism and Disposition 34: 354–360.PubMedCrossRef
38.
Warren, G.W., S.M. Poloyac, D.S. Gary, M.P. Mattson, and R.A. Blouin. 1999. Hepatic cytochrome P-450 expression in tumor necrosis factor-alpha receptor (p55/p75) knockout mice after endotoxin administration. Journal of Pharmacology and Experimental Therapeutics 288: 945–950.PubMed
39.
Siewert, E., R. Bort, R. Kluge, P.C. Heinrich, J. Castell, and R. Jover. 2000. Hepatic cytochrome P450 down-regulation during aseptic inflammation in the mouse is interleukin 6 dependent. Hepatology 32: 49–55.PubMedCrossRef
40.
Dickmann, L.J., S.K. Patel, D.A. Rock, L.C. Wienkers, and J.G. Slatter. 2011. Effects of interleukin-6 (IL-6) and an anti-IL-6 monoclonal antibody on drug-metabolizing enzymes in human hepatocyte culture. Drug Metabolism and Disposition 39: 1415–1422.PubMedCrossRef
41.
Chaluvadi, M.R., R.D. Kinloch, B.A. Nyagode, T.A. Richardson, M.J. Raynor, M. Sherman, L. Antonovic, H.W. Strobel, D.L. Dillehay, and E.T. Morgan. 2009. Regulation of hepatic cytochrome P450 expression in mice with intestinal or systemic infections of Citrobacter rodentium. Drug Metabolism and Disposition 37: 366–374.PubMedCrossRef
42.
Behnia, K., S. Bhatia, N. Jastromb, U. Balis, S. Sullivan, M. Yarmush, and M. Toner. 2000. Xenobiotic metabolism by cultured primary porcine hepatocytes. Tissue Engineering 6: 467–479.PubMedCrossRef
43.
Kojima, M., M. Sekimoto, and M. Degawa. 2010. Androgen-mediated down-regulation of CYP1A subfamily genes in the pig liver. Journal of Endocrinology 207: 203–211.CrossRefPubMed
44.
Kojima, M., M. Sekimoto, and M. Degawa. 2008. A novel gender-related difference in the constitutive expression of hepatic cytochrome P4501A subfamily enzymes in Meishan pigs. Biochemical Pharmacology 75: 1076–1082.PubMedCrossRef
45.
Hyland, K.A., D.R. Brown, and M.P. Murtaugh. 2006. Salmonella enterica serovar Choleraesuis infection of the porcine jejunal Peyer’s patch rapidly induces IL-1beta and IL-8 expression. Veterinary Immunology and Immunopathology 109: 1–11.PubMedCrossRef
46.
Nygard, A.B., C.B. Jorgensen, S. Cirera, and M. Fredholm. 2007. Selection of reference genes for gene expression studies in pig tissues using SYBR green qPCR. BMC Molecular Biology 8: 67.PubMedPubMedCentralCrossRef
Metadata
Title
The Effects of Intestinal LPS Exposure on Inflammatory Responses in a Porcine Enterohepatic Co-culture System
Authors
Erzsebet Paszti-Gere
Gabor Matis
Orsolya Farkas
Anna Kulcsar
Orsolya Palocz
Gyorgy Csiko
Zsuzsanna Neogrady
Peter Galfi
Publication date
01-02-2014
Publisher
Springer US
Published in
Inflammation / Issue 1/2014
Print ISSN: 0360-3997
Electronic ISSN: 1573-2576
DOI
https://doi.org/10.1007/s10753-013-9735-7

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