Cytokine response after stimulation of culture cells by zinc and probiotic strain

  • Miroslava ŠefcováEmail author
  • Martin Levkut
  • Katarína Bobíková
  • Viera Karaffová
  • Viera Revajová
  • Ivana Cingeľová Maruščáková
  • Mária Levkutová
  • Zuzana Ševčíková
  • Róbert Herich
  • Mikuláš Levkut


Intestinal porcine epithelial cells were used for an in vitro analysis of mRNA expression levels of inflammatory cytokines (IL-8, IL-18) and transcriptional factors (MyD88 and NF-κβ). Cells were exposed to inorganic and organic zinc sources (in two different concentrations—50 μmol/L and 100 μmol/L) alone or combined with Lactobacillus reuteri B6/1, which was also applied individually. The total exposure time was 4 h. Quantitative reverse transcriptase PCR was used to determine expression levels of the aforementioned parameters. In general, upregulation was observed; however, a decrease of some mRNA’s abundance was also determined. Differences in expression were analysed statistically using ANOVA and Tukey analyses. High relative expression was shown for IL-8, IL-18 and MyD88 in groups treated with 100 μmol/L of inorganic sources of zinc (ZnSO4) (p < 0.05), while groups treated with the organic form did not exhibit significant changes in expression. Also, 50 μmol/L of either zinc source did not significantly modify the transcriptional profile of the cytokines and transcription factors, showing that even inorganic sources, at lower concentrations, do not elicit a significant inflammatory reaction. In summary, supplementation of organic zinc source (Gly-Zn chelate) ensures that IL-8, IL-18, MyD88 and NF-κβ expression levels are not positively regulated. In contrast, inorganic sources of zinc (ZnSO4) could induce an inflammatory reaction. However, this response could be dampened if L. reuteri B6/1 is administered, showing the helpful aspect of using probiotics to modulate an inflammatory response. Conclusively, the use Gly-Zn chelate appears as an optimal alternative for Zn administration that does not compromise normal intestinal homeostasis.


Zinc glycine chelate Zinc sulphate Probiotic bacteria Cytokine Transcription factors IPEC-1 



We are very grateful to Marco Larrea-Álvarez, PhD, and to César Larrea-Álvarez for the appropriate and constructive suggestions that help to improve the manuscript. This work was supported by the Grant Agency for Science of the Slovak Republik VEGA (1/0112/18, 1/0355/19); Slovak Research and Developmental Agency (APVV-15-0165).

Supplementary material

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ESM 1 (PDF 195 kb)
11626_2019_401_MOESM2_ESM.xlsx (10 kb)
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Copyright information

© The Society for In Vitro Biology 2019

Authors and Affiliations

  • Miroslava Šefcová
    • 1
    Email author
  • Martin Levkut
    • 1
  • Katarína Bobíková
    • 1
  • Viera Karaffová
    • 1
  • Viera Revajová
    • 1
  • Ivana Cingeľová Maruščáková
    • 2
  • Mária Levkutová
    • 3
  • Zuzana Ševčíková
    • 1
  • Róbert Herich
    • 1
  • Mikuláš Levkut
    • 1
    • 4
  1. 1.Department of Pathological Anatomy and Pathological PhysiologyUniversity of Veterinary Medicine and PharmacyKošiceSlovak Republic
  2. 2.Department of Microbiology and ImmunologyUniversity of Veterinary Medicine and PharmacyKošiceSlovak Republic
  3. 3.Department of Epizootiology and ParasitologyUniversity of Veterinary Medicine and PharmacyKošiceSlovak Republic
  4. 4.Institute of NeuroimmunologySlovak Academy of ScienceBratislavaSlovak Republic

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