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.
Subject(s)
Colon/chemistry , Fibroblasts/drug effects , Galectin 3/analysis , Mucous Membrane/chemistry , Adult , Aged , Aged, 80 and over , Caco-2 Cells , Cell Line , Cells, Cultured , Colorectal Neoplasms/pathology , Culture Media, Conditioned , Diverticulitis/pathology , Epithelial Cells/chemistry , Female , HT29 Cells , Humans , Interleukin-8/analysis , Intestinal Diseases/pathology , Male , Middle Aged , NF-kappa B/metabolism , Transcription Factor AP-1/metabolismABSTRACT
To investigate whether endoscopy affects sphincter of Oddi (SO) manometry, three patients who had undergone previous cholecystectomy and had a T-tube in situ for drainage were studied. Manometry was performed using a perfused triple lumen manometry catheter (diameter 1.7 mm), which was advanced into the SO lumen through the T-tube. SO motility, baseline pressure, common bile duct pressure and duodenal pressure were monitored before and during endoscopy while the tip of the endoscope was in the mouth, esophagus (upper third, precardial), stomach and duodenum. Endoscopy and even a moderate insufflation of air necessary to pass the pylorus and inspect the papilla of Vater did not affect the parameters mentioned. Thus, ERCP manometry is a reliable method for evaluation of SO motility which is not affected by endoscopy. Duodenal pressure is a stable parameter and suitable for serving as reference pressure.
