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Induction of Nitric Oxide Synthesis by Probiotic Lactobacillus rhamnosus GG in J774 Macrophages and Human T84 Intestinal Epithelial Cells

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Abstract

Backgrounds and Aims: Probiotic Lactobacillus rhamnosus GG (LGG) has proved to be beneficial in the treatment of viral- and antibiotic-associated diarrhea but the mechanisms of action remain unknown. Nitric oxide (NO) is involved in the protective mechanisms in the gastrointestinal tract and may contribute to some of the beneficial effects of probiotics. The aim of the present study was to investigate if induction of NO synthesis is involved in the cellular actions of LGG.

Methods: NO synthesis and its regulation were measured in cultures of J774 murine macrophages and human T84 colon epithelial cells. NO production was measured as its metabolite nitrite accumulated into the culture medium. Inducible nitric oxide synthase (iNOS) protein and iNOS mRNA were detected by Western blot and RT-PCR, respectively.

Results: In J774 macrophages, LGG induced a low level production of NO in the presence of gamma interferon (IFNγ) and it was inhibited by NOS inhibitors, cycloheximide and by a NF-kappa B inhibitor pyrrolidinedithiocarbamate. Accordingly, LGG and IFNγ-stimulation increased iNOS mRNA and protein levels. T84 cells produced NO in response to LGG when first primed with a combination of IL-1β, TNFα and IFNγ. Lipoteichoic acid (LA), an antigenic structure in gram-positive bacteria, also induced NO formation in J774 cells in the presence of IFNγ suggesting that LA may be the active component in LGG.

Conclusions: LGG induces NO production in J774 macrophages and in human T84 colon epithelial cells through induction of iNOS by a mechanism involving activation of transcription factor NF-κB. Induction of iNOS and low-level synthesis of NO may be involved in the protective actions of LGG in the gastrointestinal tract.

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Authors and Affiliations

  1. The Immunopharmacological Research Group, Medical School, University of Tampere, and Tampere University Hospital, Finland

    Riku Korhonen & Hannu Kankaanranta

  2. Valio Ltd, Helsinki, Finland

    Riitta Korpela & Maija Saxelin

  3. Pediatric Research Center, Medical School, University of Tampere, Finland

    Markku Mäki

  4. Department of Pediatrics, Tampere University Hospital, Tampere, Finland

    Markku Mäki

  5. The Immunopharmacological Research Group, Medical School, University of Tampere, and Tampere University Hospital, Finland

    Eeva Moilanen

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Korhonen, R., Korpela, R., Saxelin, M. et al. Induction of Nitric Oxide Synthesis by Probiotic Lactobacillus rhamnosus GG in J774 Macrophages and Human T84 Intestinal Epithelial Cells. Inflammation 25, 223–232 (2001). https://doi.org/10.1023/A:1010971703271

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