Metabolites of Lactobacillus plantarum 2142 Prevent Oxidative Stress-Induced Overexpression of Proinflammatory Cytokines in IPEC-J2 Cell Line
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Probiotics have already proven beneficial effects in the treatment of several intestinal infections, but the underlying mechanisms how the probiotics can affect responses of porcine IPEC-J2 enterocytes to oxidative stress remained to be elucidated. The immunmodulatory effect of five bacterial strains (Lactobacillus plantarum 2142, Lactobacillus casei Shirota, Bifidobacterium animalis subsp. lactis BB-12, Bacillus amyloliquefaciens CECT 5940 and Enterococcus faecium CECT 4515) on 1 mM peroxide-triggered upregulation of interleukin (IL)-8 and tumor necrosis factor alpha (TNF-α) level was screened by q RT-PCR. Our data revealed that spent culture supernatant (SCS) of L. plantarum 2142 had significant lowering effect on IL-8 and TNF-α level with concomitant promoting activity on protective Hsp70 gene expression. According to our results, lactic acid (racemic, d- and l-lactic acid) and acetic acid produced by lactobacilli had no protective effect in quenching upregulation of proinflammatory cytokines. Furthermore, L. plantarum 2142-specific supernatant peptides were detected by gel electrophoresis and capillary zone electrophoresis.
KEY WORDSIPEC-J2 probiotics oxidative stress proinflammatory cytokines Hsp70
The research described here has been supported by the Hungarian Scientific Research Fund (grant OTKA nos. 76133 and 100701). We are indebted to Dr. Jody Gookin and Dr. Stephen Stauffer, Department of Clinical Sciences, College of Veterinary Medicine, North Carolina State University, Raleigh, NC, USA for providing IPEC-J2 cells and for the valuable advice on handling them. PCR product sequencing support from Dr. Balázs Gereben (Institute of Experimental Medicine of the Hungarian Academy of Sciences, Laboratory of Endocrine Neurobiology) is also acknowledged. We also would like to thank Dr. Éva Gelencsér, Dr. Emőke Németh-Szerdahelyi and Katalin Háder-Sólyom (Central Food Research Institute, Food Safety Department, Unit of Biology, Budapest, Hungary) for their extensive support in peptide electrophoretic studies.
- 6.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 and Technology. doi: 10.1016/j.tifs.2011.03.010.
- 9.Karczewski, J., F.J. Troost, I. Konings, J. Dekker, M. Kleerebezem, R.J.M. Brummer, and J.M. Wells. 2010. Regulation of human epithelial tight junction proteins by Lactobacillus plantarum in vivo and protective effects on the epithelial barrier. Translational Physiology 298: G851–G859.Google Scholar
- 12.Ewaschuk, J.B., H. Diaz, L. Meddings, B. Diederichs, A. Dmytrash, J. Backer, M. Looijer-van Langen, and K.L. Madsen. 2008. Secreted bioactive factors from Bifidobacterium infantis enhance epithelial cell barrier function. AJP-Gastrointestinal and Liver Physiology 295: 1025–1034.CrossRefGoogle Scholar
- 15.Hobbie, S., L.M. Chen, R.J. Davis, and J.E. Galán. 1997. Involvement of mitogen-activated protein kinase pathways in the nuclear responses and cytokine production induced by Salmonella typhimurium in cultured intestinal epithelial cells. Journal of Immunology 159: 5550–5559.Google Scholar
- 16.McCormick, B.A., C.A. Parkos, S.P. Colgan, D.K. Carnes, and J.L. Madara. 1998. Apical secretion of a pathogen-elicited epithelial chemoattractant activity in response to surface colonization of intestinal epithelia by Salmonella typhimurium. Journal of Immunology 160: 455–466.Google Scholar
- 17.Wilson, M., R. Seymour, and B. Henderson. 1998. Bacterial perturbation of cytokine networks. Infection and Immunology 66: 2401–2409.Google Scholar
- 18.Lang, A., M. Lahav, E. Sakhnini, I. Barshack, H.H. Fiddler, B. Avidan, E. Bardan, R.R. Hershkowiz, S. Bar-Meir, and Y. Chowers. 2004. Allicin inhibits spontaneous and TNF-alpha induced secretion of proinflammatory cytokines and chemokines from intestinal epithelial cells. Clinical Nutrition 23: 1199–1208.PubMedCrossRefGoogle Scholar
- 19.Chowers, Y., L. Cahalon, M. Lahav, H. Schor, R. Tal, S. Bar-Meir, and M. Levite. 2000. Somatostatin through its specific receptor inhibits spontaneous and TNF-alpha- and bacteria-induced IL-8 and IL-1 beta secretion from intestinal epithelial cells. Journal of Immunology 165: 2955–2961.Google Scholar
- 20.Yamamoto, K., R. Kushima, O. Kisaki, Y. Fujiyama, and H. Okabe. 2003. Combined effect of hydrogen peroxide induced oxidative stress and IL-1 on IL-8 production in CaCo-2 cells (a human colon carcinoma cell line) and normal intestinal epithelial cells. Inflammation 27: 123–128.PubMedCrossRefGoogle Scholar
- 22.Coconnier, M.H., V. Lievin, M. Lorrot, and A.L. Servin. 2000. Antagonistic activity of Lactobacillus acidophilus LB against intracellular Salmonella enterica serovar Typhimurium infecting human enterocyte-like Caco-2/TC-7 cells. Applied and Environmental Microbiology 66: 1152–1157.PubMedCrossRefGoogle Scholar
- 23.Candela, M., F. Perna, P. Carnevali, B. Vitali, R. Ciati, P. Gionchetti, F. Rizello, M. Campieri, and P. Brigidi. 2008. Interaction of probiotic Lactobacillus and Bifidobacterium strains with human intestinal epithelial cells: adhesion properties, competition against enteropathogens and modulation of IL-8 production. International Journal of Food Microbiology 125: 286–292.PubMedCrossRefGoogle Scholar
- 30.Musch, M.W., M.J. Ciancio, K. Sarge, and E.B. Chang. 1996. Induction of heat shock protein 70 protects intestinal epithelial IEC-18 cells from oxidant and thermal injury. American Journal of Physiology. Cell Physiology 270(2): C429–C436.Google Scholar
- 32.Nemeth, E., S. Fajdiga, J. Malago, J. Koninkx, P. Tooten, and J. van Dijk. 2006. Inhibition of Salmonella-induced IL-8 synthesis and expression of Hsp70 in enterocyte-like Caco-2 cells after exposure to non-starter lactobacilli. International Journal of Food Microbiology 112(3): 266–274.PubMedCrossRefGoogle Scholar
- 36.Chon, H., B. Choi, G. Jeong, E. Lee, and S. Lee. 2010. Suppression of proinflammatory cytokine production by specific metabolites of Lactobacillus plantarum 10hk2 via inhibiting NF-κB and p38 MAPK expressions. Comparative Immunology, Microbiology & Infection Diseases 33: 41–49.CrossRefGoogle Scholar
- 44.Watanabe, T., H. Nishio, T. Tanigawa, H. Yamagami, H. Okazaki, K. Watanabe, K. Tominaga, Y. Fujiwara, N. Oshitani, T. Asahara, K. Nomoto, K. Higuchi, K. Takeuchi, and T. Arakawa. 2009. Probiotic Lactobacillus casei strain Shirota prevents indomethacin-induced small intestinal injury: involvement of lactic acid. AJP-Gastrointestinal and Liver Physiology 297: 506–513.CrossRefGoogle Scholar
- 45.Dietl, K., K. Renner, K. Dettmer, B. Timischl, K. Eberhart, C. Dorn, C. Hellerbrand, M. Kastenberger, L.A. Kunz-Schughart, P.J. Oefner, R. Andreesen, E. Gottfried, and M.P. Kreutz. 2010. Lactic acid and acidification inhibit TNF secretion and glycolysis of human monocytes. Journal of Immunology 184: 1200–1209.CrossRefGoogle Scholar
- 51.Schierack, P., M. Nordhoff, M. Pollmann, K.D. Weyrauch, S. Amasheh, U. Lodemann, J. Jores, B. Tachu, S. Kleta, A. Blikslager, K. Tedin, and L.H. Wieler. 2006. Characterization of a porcine intestinal epithelial cell line for in vitro studies of microbial pathogenesis in swine. Histochemistry and Cell Biolology 125: 293–305.CrossRefGoogle Scholar
- 52.Paszti-Gere, E., E. Csibrik-Nemeth, K. Szeker, Cs Jakab, and P. Galfi. 2011. Acute oxidative stress affects IL-8 and TNF-α expression in IPEC-J2 porcine epithelial cells. Inflammation DOI:. doi: 10.1007/s10753-011-9403-8.
- 54.Xu, Q., T. Yajima, W. Li, K. Saito, Y. Ohshima, and Y. Yoshikai. 2006. Levan (β-2, 6-fructan), a major fraction of fermented soybean mucilage, displays immunostimulating properties via Toll-like receptor 4 signalling: induction of interleukin-12 production and suppression of T-helper type 2 response and immunoglobulin E production. Clinical and Experimental Allergy 36(1): 94–101.PubMedCrossRefGoogle Scholar