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Osteoprotegerin Exerts Its Pro-inflammatory Effects Through Nuclear Factor-κB Activation

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Abstract

Background

Osteoprotegerin (OPG), a soluble member of the tumor necrosis factor (TNF) receptor super-family, is a key factor inhibiting the differentiation and activation of osteoclasts. It has recently been implicated as a disease marker for inflammatory bowel disease (IBD) yet its role in the intestinal epithelial inflammatory response remains unknown.

Aim

The primary objective of this study was to investigate whether OPG has a role in intestinal inflammation and a potential role in IBD pathogenesis.

Methods

Caco-2 and HT-29 cells were grown in vitro to confluence on culture-permeable supports and then co-cultured with either TNF-α or OPG. After exposure to either TNF-α or OPG, interleukin (IL)-8 protein and mRNA levels were evaluated. Ussing chamber, western blotting, real-time polymerase chain reaction, and immunofluorescence were used to further investigate the effect of OPG on intestinal barrier integrity and function.

Results

Similar to TNF-α, treatment of monolayers with OPG caused increased monolayer permeability and diminished tight junction function and integrity, with loss of tight junction proteins from cell membranes. This was accompanied by elevated IL-8 protein and gene levels (P < 0.05). Western blotting also revealed that OPG, similar to TNF-α, induced NF-κB activation, as shown by inhibition of NF-κB kinase subunit-α phosphorylation.

Conclusions

These results indicate that OPG has pro-inflammatory properties because it induces gut barrier dysfunction and secretion of other pro-inflammatory cytokines. These results also provide evidence that OPG is likely to exert its pro-inflammatory effects through NF-κB activation and may potently contribute to IBD pathogenesis.

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Acknowledgments

This work was supported by the National Health and Medical Research Council, Australia (NHMRC, grant number 510230). Laboratory investigations were performed in the Westfield Research Laboratories.

Conflict of interest

None.

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Authors and Affiliations

  1. School of Women’s and Children’s Health, University of New South Wales, High Street, Randwick, Sydney, NSW, 2031, Australia

    Lily Nahidi, Steven T. Leach, Daniel A. Lemberg & Andrew S. Day

  2. Department of Gastroenterology, Sydney Children’s Hospital, High Street, Randwick, Sydney, NSW, 2031, Australia

    Daniel A. Lemberg

  3. Department of Paediatrics, University of Otago, Christchurch, Christchurch, 8140, New Zealand

    Andrew S. Day

  4. Department of Paediatric Gastroenterology, Christchurch Hospital, Christchurch, 8140, New Zealand

    Andrew S. Day

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  1. Lily Nahidi
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  2. Steven T. Leach
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Corresponding author

Correspondence to Steven T. Leach.

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Nahidi, L., Leach, S.T., Lemberg, D.A. et al. Osteoprotegerin Exerts Its Pro-inflammatory Effects Through Nuclear Factor-κB Activation. Dig Dis Sci 58, 3144–3155 (2013). https://doi.org/10.1007/s10620-013-2851-2

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  • DOI: https://doi.org/10.1007/s10620-013-2851-2

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