D-alanine Inhibits Murine Intestinal Inflammation by Suppressing IL-12 and IL-23 Production in Macrophages

J Crohns Colitis. 2024 Jun 3;18(6):908-919. doi: 10.1093/ecco-jcc/jjad217.

Abstract

Background and aims: Free D-amino acids, which have different functions from L-amino acids, have recently been discovered in various tissues. However, studies on the potential interactions between intestinal inflammation and D-amino acids are limited. We examined the inhibitory effects of D-alanine on the pathogenesis of intestinal inflammation.

Methods: We investigated serum D-amino acid levels in 40 patients with ulcerative colitis and 34 healthy volunteers. For 7 days [d], acute colitis was induced using dextran sulphate sodium in C57BL/6J mice. Plasma D-amino acid levels were quantified in mice with dextran sulphate sodium-induced colitis, and these animals were administered D-alanine via intraperitoneal injection. IFN-γ, IL-12p35, IL-17A, and IL-23p19 mRNA expression in the colonic mucosa was measured using real-time polymerase chain reaction [PCR]. In vitro proliferation assays were performed to assess naïve CD4+ T cell activation under Th-skewing conditions. Bone marrow cells were stimulated with mouse macrophage-colony stimulating factor to generate mouse bone marrow-derived macrophages.

Results: Serum D-alanine levels were significantly lower in patients with ulcerative colitis than in healthy volunteers. Dextran sulphate sodium-treated mice had significantly lower plasma D-alanine levels than control mice. D-alanine-treated mice had significantly lower disease activity index than control mice. IFN-γ, IL-12p35, IL-17A, and IL-23p19 mRNA expression levels were significantly lower in D-alanine-administered mice than in control mice. D-alanine suppressed naïve T cell differentiation into Th1 cells in vitro, and inhibited the production of IL-12p35 and IL-23p19 in bone marrow-derived macrophages.

Conclusions: Our results suggest that D-alanine prevents dextran sulphate sodium-induced colitis in mice and suppresses IL-12p35 and IL-23p19 production in macrophages.

Keywords: D-alanine; D-amino acid; ulcerative colitis.

MeSH terms

  • Adult
  • Alanine* / pharmacology
  • Animals
  • Case-Control Studies
  • Colitis, Ulcerative* / chemically induced
  • Colitis, Ulcerative* / drug therapy
  • Colitis, Ulcerative* / immunology
  • Colitis, Ulcerative* / metabolism
  • Colitis, Ulcerative* / pathology
  • Dextran Sulfate*
  • Disease Models, Animal
  • Female
  • Humans
  • Interferon-gamma / metabolism
  • Interleukin-12 / metabolism
  • Interleukin-12 Subunit p35 / metabolism
  • Interleukin-17 / metabolism
  • Interleukin-23 Subunit p19 / metabolism
  • Interleukin-23* / metabolism
  • Intestinal Mucosa / drug effects
  • Intestinal Mucosa / metabolism
  • Intestinal Mucosa / pathology
  • Macrophages* / drug effects
  • Macrophages* / metabolism
  • Male
  • Mice
  • Mice, Inbred C57BL*
  • Middle Aged
  • RNA, Messenger / metabolism
  • Young Adult

Substances

  • Dextran Sulfate
  • Alanine
  • Interleukin-23
  • Interleukin-12
  • Interferon-gamma
  • Interleukin-17
  • RNA, Messenger
  • Interleukin-12 Subunit p35
  • Interleukin-23 Subunit p19