Soluble galectin-3 is a strong, colonic epithelial-cell-derived, lamina propria fibroblast-stimulating factor.

Abstract

BACKGROUND:

Colonic lamina propria fibroblasts (CLPFs) play an important role in the pathogenesis of fibrosis and strictures in Crohn's disease.

AIM:

To identify colonic epithelial cell (CEC)-derived factors that activate CLPFs.

METHODS:

Primary human CECs and CLPFs were isolated from control mucosa and interleukin 8 (IL8) of CLPF cultures was quantified by ELISA. Activation of nuclear factor kappaB (NF-kappaB) was shown, and translocation of NF-kappaB was inhibited by a dominant-negative IkappaB-expressing adenovirus. The major CLPF-activating and IL8 inducing protein was purified using fast-performance liquid chromatography (HiPrep 16/60 Sephacryl S-200 High Resolution Column) and sodium dodecyl sulphate gel electrophoresis.

RESULTS:

A considerable increase in IL8 secretion by CLPFs cultured in CEC-conditioned media compared with that in unconditioned media (155.00 (10.00) pg/microg v 1.434 (0.695) pg/microg) was found. The effect of CEC-conditioned media on CLPF IL8 secretion was NF-kappaB dependent. A protein or DNA array confirmed the involvement of NF-kappaB and activator protein-1. Purification of a candidate band isolated with the use of sodium dodecyl sulphate-polyacrylamide gel electrophoresis and subsequent sequencing showed soluble galectin-3 to be a strong CLPF-activating factor. Depletion of galectin-3 from conditioned media by immunoprecipitation abolished the CLPF stimulatory effect.

CONCLUSIONS:

Using a classical biochemical approach, soluble galectin-3 was identified as a strong activator of CLPFs produced by CEC. Galectin-3 induced NF-kappaB activation and IL8 secretion in these cells and may be a target for future therapeutic approaches to reduce or avoid stricture formation.

• Authors: E Lippert, W Falk, F Bataille, T Kaehne, M Naumann, M Goeke, H Herfarth, J Schoelmerich, G Rogler
• Journal: Gut (Gut) Vol. 56 Issue 1 Pg. 43-51 (Jan 2007) ISSN: 0017-5749 [Print] England
• PMID: 16709662 (Publication Type: Journal Article)

Chemical References

• Culture Media, Conditioned
• Galectin 3
• Interleukin-8
• NF-kappa B
• Transcription Factor AP-1

Topics

• Adult
• Aged
• Aged, 80 and over
• Caco-2 Cells
• Cell Line
• Cells, Cultured
• Colon (chemistry)
• Colorectal Neoplasms (pathology)
• Culture Media, Conditioned
• Diverticulitis (pathology)
• Epithelial Cells (chemistry)
• Female
• Fibroblasts (drug effects)
• Galectin 3 (analysis)
• HT29 Cells
• Humans
• Interleukin-8 (analysis)
• Intestinal Diseases (pathology)
• Male
• Middle Aged
• Mucous Membrane (chemistry)
• NF-kappa B (metabolism)
• Transcription Factor AP-1 (metabolism)