γ-Glutamyl cysteine and γ-glutamyl valine inhibit TNF-α signaling in intestinal epithelial cells and reduce inflammation in a mouse model of colitis via allosteric activation of the calcium-sensing receptor

Biochim Biophys Acta. 2015 May;1852(5):792-804. doi: 10.1016/j.bbadis.2014.12.023. Epub 2015 Jan 3.

Abstract

Background: The extracellular calcium-sensing receptor (CaSR) is distributed throughout the gastrointestinal tract, and its activation has been shown to promote intestinal homeostasis, suggesting that CaSR may be a promising target for novel therapies to prevent chronic intestinal inflammation such as inflammatory bowel disease (IBD). The γ-glutamyl dipeptides γ-glutamyl cysteine (γ-EC) and γ-glutamyl valine (γ-EV) are dietary flavor enhancing compounds, and have been shown to activate CaSR via allosteric ligand binding. The aim of this study was to examine the anti-inflammatory effects of γ-EC and γ-EV in vitro in intestinal epithelial cells and in a mouse model of intestinal inflammation.

Results: In vitro, treatment of Caco-2 cells with γ-EC and γ-EV resulted in the CaSR-mediated reduction of TNF-α-stimulated pro-inflammatory cytokines and chemokines including IL-8, IL-6, and IL-1β, and inhibited phosphorylation of JNK and IκBα, while increasing expression of IL-10. In vivo, using a mouse model of dextran sodium sulfate (DSS)-induced colitis, γ-EC and γ-EV treatment ameliorated DSS-induced clinical signs, weight loss, colon shortening and histological damage. Moreover, γ-EC and γ-EV reduced the expression of TNF-α, IL-6, INF-γ, IL-1β, and IL-17, and increased the expression of IL-10 in the colon, in a CaSR-dependent manner. The CaSR-mediated anti-inflammatory effects of γ-EC were abrogated in β-arrestin2 knock-down Caco-2 cells, and involvement of β-arrestin2 was found to inhibit TNF-α-dependent signaling via cross-talk with the TNF-α receptor (TNFR).

Conclusions: Thus CaSR activation by γ-EC and γ-EV can aid in maintaining intestinal homeostasis and reducing inflammation in chronic inflammatory conditions such as IBD.

Keywords: Calcium-sensing receptor (CaSR); Dextran sodium sulfate (DSS) colitis; Inflammatory bowel disease (IBD); Intestinal epithelium; Tumor necrosis factor (TNF); γ-Glutamyl dipeptides.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Allosteric Regulation / drug effects
  • Animals
  • Arrestins / genetics
  • Arrestins / metabolism
  • Blotting, Western
  • Caco-2 Cells
  • Colitis / chemically induced
  • Colitis / metabolism
  • Colitis / prevention & control*
  • Cytokines / metabolism
  • Dextran Sulfate
  • Dipeptides / pharmacology*
  • Epithelial Cells / drug effects*
  • Epithelial Cells / metabolism
  • Female
  • Humans
  • Inflammation Mediators / metabolism
  • Intestines / cytology
  • JNK Mitogen-Activated Protein Kinases / metabolism
  • Mice, Inbred BALB C
  • Phosphorylation / drug effects
  • RNA Interference
  • Receptors, Calcium-Sensing / metabolism*
  • Receptors, Tumor Necrosis Factor / metabolism
  • Signal Transduction / drug effects
  • Tumor Necrosis Factor-alpha / metabolism
  • Tumor Necrosis Factor-alpha / pharmacology*
  • beta-Arrestins

Substances

  • Arrestins
  • Cytokines
  • Dipeptides
  • Inflammation Mediators
  • Receptors, Calcium-Sensing
  • Receptors, Tumor Necrosis Factor
  • Tumor Necrosis Factor-alpha
  • beta-Arrestins
  • gamma-glutamylvaline
  • Dextran Sulfate
  • JNK Mitogen-Activated Protein Kinases
  • gamma-glutamylcysteine