Skip to main content
SpringerLink
Log in

Zn2+ and l-isoleucine induce the expressions of porcine β-defensins in IPEC-J2 cells

  • Published:
Molecular Biology Reports Aims and scope Submit manuscript

Abstract

The β-defensins, expressed in epithelial cells of multiple tissues including intestine, play a critical role in the mammalian innate immunity. However, it is little known about the role of functional nutrients in the regulation of porcine β-defensins’ expressions in intestinal epithelial cells. The present study was conducted to determine the hypothesis that zinc and l-isoleucine regulate the expressions of porcine β-defensins in IPEC-J2 cells. Cells were cultured in DMEM/F12 medium containing supplemental 0–500 μg/mL l-isoleucine or 0–500 μmol/mL zinc sulfate that was used to increase the concentration of Zn2+ in the medium. At 12 h after the treatment by the appropriate concentrations of l-isoleucine or Zn2+, the mRNA and protein expressions of porcine β-defensin 1, 2 and 3 were increased (P < 0.05), and reached their maximum after treatment with 25 or 100 μmol/mL zinc sulfate and 25 or 50 μg/mL isoleucine (P < 0.05). These results suggested that both Zn2+ and l-isoleucine could induce β-defensins’ expressions in porcine intestinal epithelial cells.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2

Similar content being viewed by others

References

  1. Ganz T (2003) Defensins: antimicrobial peptides of innate immunity. Nat Rev Immunol 3:710–720

    Article  PubMed  CAS  Google Scholar 

  2. Froy O, Gurevitz M (2003) Arthropod and mollusk defensins-evolution by exon-shuffling. Trends Genet 19:684–687

    Article  PubMed  CAS  Google Scholar 

  3. Evans EW, Harmon BG (1995) A review of antimicrobial peptides: defensins and related cationic peptides. Vet Clin Pathol 24:109–116

    Article  PubMed  Google Scholar 

  4. Kaiser V, Diamond G (2000) Expression of mammalian defensin genes. J Leukoc Biol 68:779–784

    PubMed  CAS  Google Scholar 

  5. Qi S, Chen J, Guo R, Yu B, Chen D (2009) β-defensins gene expression in tissues of the crossbred and Tibetan pigs. Livest Sci 123:161–168

    Article  Google Scholar 

  6. Zhang G, Hiraiwa H, Yasue H, Wu H, Ross CR, Troyer D, Blecha F (1999) Cloning and characterization of the gene for a new epithelial beta-defensin. Genomic structure, chromosomal localization, and evidence for its constitutive expression. J Biol Chem 274:24031–24037

    Article  PubMed  CAS  Google Scholar 

  7. Veldhuizen EJ, van Dijk A, Tersteeg MH, Kalkhove SI, van der Meulen J, Niewold TA, Haagsman HP (2007) Expression of beta-defensins pBD-1 and pBD-2 along the small intestinal tract of the pig: lack of upregulation in vivo upon Salmonella typhimurium infection. Mol Immunol 44:276–283

    Article  PubMed  CAS  Google Scholar 

  8. Fehlbaum P, Rao M, Zasloff M, Anderson GM (2000) An essential amino acid induces epithelial beta-defensin expression. Proc Natl Acad Sci USA 97:12723–12728

    Article  PubMed  CAS  Google Scholar 

  9. Harder J, Meyer-Hoffert U, Wehkamp K, Schwichtenberg L, Schröder JM (2004) Differential gene induction of human beta-defensins (hBD-1, -2 -3, and -4) in keratinocytes is inhibited by retinoic acid. J Invest Dermatol 123:522–529

    Article  PubMed  CAS  Google Scholar 

  10. Sherman H, Chapnik N, Froy O (2006) Albumin and amino acids upregulate the expression of human beta-defensin 1. Mol Immunol 43:1617–1623

    Article  PubMed  CAS  Google Scholar 

  11. Liu F, Abiko Y, Nishimura M, Kusano K, Shi S, Kaku T, Lin F (2001) Expression of inflammatory cytokines and beta-defensin 1 mRNAs in porcine epithelial rests of Malassez in vitro. Med Electron Microsc 34:174–178

    Article  PubMed  CAS  Google Scholar 

  12. Tarver AP, Clark DP, Diamond G, Russell JP, Erdjument-Bromage H, Tempst P, Cohen KS, Jones DE, Sweeney RW, Wines M, Hwang S, Bevins CL (1998) Enteric beta-defensin: molecular cloning and characterization of a gene with inducible intestinal epithelial cell expression associated with Cryptosporidium parvum infection. Infect Immun 66:1045–1056

    PubMed  CAS  Google Scholar 

  13. Mao X, Qi S, Yu B, Huang Z, Chen H, Mao Q, Han G, Chen D (2012) Dietary l-arginine supplementation enhances porcine β-defensins gene expression in some tissues of weaned pigs. Livest Sci. doi:10.1016/j.livsci.2012.05.016

    Google Scholar 

  14. Aschkenasy A (1975) Dietary proteins and amino acids in leucopoiesis: recent hematological and immunological data. World Rev Nutr Diet 21:151–197

    PubMed  CAS  Google Scholar 

  15. Konno Y, Ashida T, Inaba Y, Ito T, Tanabe H, Maemoto A, Ayabe T, Mizukami Y, Fujiya M, Kohgo Y (2012) Isoleucine, an essential amino acid, induces the expression of human β defensin 2 through the activation of the G-protein coupled receptor-ERK pathway in the intestinal epithelia. Food Nutr Sci 3:548–555

    Article  CAS  Google Scholar 

  16. Wintergerst ES, Maggini S, Hornig DH (2007) Contribution of selected vitamins and trace elements to immune function. Ann Nutr Metab 51:301–323

    Article  PubMed  CAS  Google Scholar 

  17. Shankar AH, Prasad AS (1998) Zinc and immune function: the biological basis of altered resistance to infection. Am J Clin Nutr 68:447S–463S

    PubMed  CAS  Google Scholar 

  18. Ibs K, Rink L (2003) Zinc-altered immune function. J Nutr 133:1452S–1456S

    PubMed  CAS  Google Scholar 

  19. Schierack P, Nordhoff M, Pollmann M, Weyrauch KD, Amasheh S, Lodemann U, Jores J, Tachu B, Kleta S, Blikslager A, Tedin K, Wieler LH (2006) Characterization of a porcine intestinal epithelial cell line for in vitro studies of microbial pathogenesis in swine. Histochem Cell Biol 125:293–305

    Article  PubMed  CAS  Google Scholar 

  20. Tan B, Yin Y, Kong X, Li P, Li X, Gao H, Li X, Huang R, Wu G (2010) l-Arginine stimulates proliferation and prevents endotoxin-induced death of intestinal cells. Amino Acids 38:1227–1235

    Article  PubMed  CAS  Google Scholar 

  21. Pfaffl MW (2001) A new mathematical model for relative quantification in real-time PR-PCR. Nucleic Acids Res 29:2002–2007

    Article  Google Scholar 

  22. Saadia H, Schein M, McFarlane C, Boffard KD (1990) Gut barrier function and the surgeon. Br J Surg 77:487–492

    Article  PubMed  CAS  Google Scholar 

  23. Mao X, Zeng X, Qiao S, Wu G, Li D (2011) Specific roles of threonine in intestinal mucosal integrity and barrier function. Front Biosci (Elite Ed) 3:1192–1200

    Google Scholar 

  24. Huttner KM, Bevins CL (1999) Antimicrobial peptides as mediators of epithelial host defense. Pediatr Res 45:785–794

    Article  PubMed  CAS  Google Scholar 

  25. Ingalls RR, Heine H, Lien E, Yoshimura A, Golenbock D (1999) Lipopolysaccharide recognition CD14 and lipopolysaccharide receptors. Infect Dis Clin North Am 13:341–353

    Article  PubMed  CAS  Google Scholar 

  26. Czop JK, Kay J (1991) Isolation and characterization of beta-glucan receptors on human mononuclear phagocytes. J Exp Med 173:1511–1520

    Article  PubMed  CAS  Google Scholar 

  27. Heeg K, Sparwasser T, Lipford GB, Häcker H, Zimmermann S, Wagner H (1998) Bacterial DNA as an evolutionary conserved ligand signalling danger of infection to immune cells. Eur J Clin Microbiol Infect Dis 17:464–469

    PubMed  CAS  Google Scholar 

  28. Moon SK, Lee HY, Li JD, Nagura M, Kang SH, Chun YM, Linthicum FH, Ganz T, Andalibi A, Lim DJ (2002) Activation of a Src-dependent Raf-MEK1/2-ERK signaling pathway is required for IL-1alpha-induced upregulation of beta-defensin 2 in human middle ear epithelial cells. Biochim Biophys Acta 1590:41–51

    Article  PubMed  CAS  Google Scholar 

  29. Kao CY, Chen Y, Thai P, Wachi S, Huang F, Kim C, Harper RW, Wu R (2004) IL-17 markedly up-regulates beta-defensin-2 expression in human airway epithelium via JAK and NF-kappaB signaling pathways. J Immunol 173:3482–3491

    PubMed  CAS  Google Scholar 

  30. Yang D, Chertov O, Bykovskaia SN, Chen Q, Buffo MJ, Shogan J, Anderson M, Schröder JM, Wang JM, Howard OMZ, Oppenheim JJ (1999) β-Defensins: linking innate and adaptive immunity through dendritic and T cell CCR6. Science 286:525–528

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgments

This study was financially supported by the Program of the “Eleventh Five-year Plan” for Science and Technology Research of China (2007BAD51B04) and the earmarked fund for Modern Agro-industry Technology Research System of China (CARS-36). We are grateful to Professor Per Torp Sangild from Copenhagen University for his donation of IPEC-J2 cells.

Author information

Authors and Affiliations

  1. Animal Nutrition Institute, Sichuan Agricultural University, Ya’an, Sichuan, 625014, China

    Xiangbing Mao, Sharina Qi, Bing Yu, Jun He, Jie Yu & Daiwen Chen

  2. Key Laboratory of Animal Disease-Resistance Nutrition, Ministry of Education, Sichuan Agricultural University, Ya’an, Sichuan, 625014, China

    Xiangbing Mao, Sharina Qi, Bing Yu, Jun He, Jie Yu & Daiwen Chen

Authors
  1. Xiangbing Mao
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Sharina Qi
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Bing Yu
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Jun He
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Jie Yu
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Daiwen Chen
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Daiwen Chen.

Additional information

Xiangbing Mao and Sharina Qi contributed equally to this work.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Mao, X., Qi, S., Yu, B. et al. Zn2+ and l-isoleucine induce the expressions of porcine β-defensins in IPEC-J2 cells. Mol Biol Rep 40, 1547–1552 (2013). https://doi.org/10.1007/s11033-012-2200-0

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11033-012-2200-0

Keywords

Search

Navigation