45 / 172
e106
Abstracts of the 22
nd
National Congress of Digestive Diseases / Digestive and Liver Disease 48S2 (2016) e67–e231
regulation of IFN-gamma, TNF-Alpha and IL-17A and were largely
protected against TNBS- and transfer-colitis.
Conclusions:
Inhibition of ATPase with a specific and selective
compound induces intestinal T cell death thereby ameliorating
colitis.
OC.09.8
INTERLEUKIN-34 INDUCES CC-CHEMOKINE LIGAND 20 IN GUT
EPITHELIAL CELLS
Franzè E.*
1
, Marafini I.
1
, De Simone V.
1
, Monteleone I.
1
, Caprioli F.
2
,
Colantoni A.
1
, Ortenzi A.
1
, Crescenzi F.
1
, Sileri P.
3
, Sica G.
3
, Rossi P.
3
,
Pallone F.
1
, Monteleone G.
1
1
Department of Systems Medicine, University of Rome “TOR VERGATA
”, Roma, Italy,
2
Department of Pathophysiology and Transplantation,
University of Milan, milano, Italy,
3
Department of Surgery, University
“TOR VERGATA ” of Rome, Roma, Italy
Background and aim:
Production of chemokines by intestinal
epithelial cells is a key step in the amplification of the destructive
immune-inflammatory response in patients with inflammatory
bowel diseases (IBD). In this study, we examined whether intestinal
epithelial cells express macrophage colony-stimulating factor
receptor 1 (M-CSFR-1), the functional receptor of interleukin-34 (IL-
34), a cytokine that is over-produced in IBD and supposed to sustain
inflammatory pathways.
Material and methods:
M-CSFR-1 expression was evaluated in
intestinal samples of IBD patients, controls and in colon epithelial
cell lines by real-time PCR, immunohistochemistry and Western
blotting. DLD-1 cells were stimulated with IL-34 in the presence
or absence of MAP kinase inhibitors, and chemokine induction
was assessed by real-time PCR and enzyme-linked immunosorbent
assay (ELISA) and MAP kinase activation was monitored by Western
blotting. The effect of a neutralizing IL-34 antibody on CCL20
synthesis was tested in ex vivo organ cultures of IBD mucosal
explants.
Results:
Enhanced expression of M-CSFR-1 RNA transcripts was
seen in inflamed mucosa of IBD patients as compared to controls.
Immunohistochemical analysis confirmed up-regulation of
M-CSFR-1 in IBD and showed that both epithelial and lamina
propria mononuclear cells expressed this receptor. Stimulation of
DLD-1 with IL-34 increased CCL20 production through an ERK1/2-
dependent mechanism. Consistently, treatment of IBD explants with
anti-IL-34 reduced CCL20 production.
Conclusions:
These data show that intestinal epithelial cells are a
target of IL-34 and suggest that this cytokine contributes to mediate
the cross talk between epithelial cells and immune cells in IBD.
OC.09.9
PROTECTIVE EFFECT OF INULIN ON LPS-INDUCED INTESTINAL
SMOOTH MUSCLE IMPAIRMENT: A PROTEOMIC APPROACH
Guarino M.*
1
, Vannini C.
2
, Altomare A.
1
, Barera S.
2
, Locato V.
3
,
Cocca S.
1
, Arrigoni G.
2
, Alloni R.
4
, De Gara L.
3
, Cicala M.
1
1
Campus Bio-Medico, Gastroenterology Unit, Rome, Italy,
2
University of
Insubria, Department of Biotechnology and Life Science,, Varese, Italy,
3
Campus Bio-Medico University, Food Sciences and Human Nutrition
Unit,, Rome, Italy,
4
Campus Bio-Medico University, Surgey Unit, Rome,
Italy
Background and aim:
Fructans, such as inulin, are dietary fibers
which stimulate gastro-intestinal function acting as prebiotics. We
recently demonstrated the protective effect of inulin on LPS-induced
damage of colonic smooth muscle in an ex vivo experimental model,
which seems to be related to presence of oxidative stress. In the
present study, the protective role of inulin against LPS-induced
oxidative stress was evaluated on colonic mucosa using a proteomic
approach.
Material and methods:
Human colonic mucosa and submucosa,
obtained from disease-free margins of resected segments for cancer,
were sealed between two chambers containing Krebs solution,
with the luminal side of the mucosa overlayed with 5 ml of Krebs,
or 100 µg/mL LPS solution, or 100 µg/mL LPS +100 mg/mL inulin
Fructafit IQ (LPS+INU). The biological system was kept oxygenated
for 30 min at 37°C. iTRAQ based analysis was used to separate and
compare the total soluble proteomes from human colonic mucosa
and submucosa treated. Each sample was labelled by one of four
reagents of the iTRAQ 4-plex and then combined into one aliquote.
Triplicate labelling were performed, which showed a high level of
reproducibility.
Results:
Inulin exposure was able to restore, in human colonic
mucosa, the LPS-dependent alteration of some proteins involved in
the host response and in the intestinal smooth muscle contraction
(ZG16, CALM1/MLCK/MYL signaling pathway) and to reduce the
upregulation of two proteins involved in the radical-mediated
oxidative stress induced by LPS (APEX1, CCT7). Moreover the
administration of inulin entails a higher level of some detoxification
enzymes (MT2A, GSTK1, and UGT2B4) with respect to LPS treatment.
Consistently inulin exposure to colonic mucosa and submucosa was
also able to restore the LPS-induced alteration of intestinal smooth
muscle contraction as well as it was able to prevent the oxidative
damages of LPS-exposed tissues.
Conclusions:
Our preliminary data suggest that the exposure of
colonic mucosa to inulin is able to prevent LPS-dependent altered
expression of some key proteins which promote intestinal motility
and the host response, reducing the radical-mediated oxidative
stress.
OC.10 Endoscopy 3
OC.10.1
DILATION-ASSISTED STONE EXTRACTION AS ALTERNATIVE,
EFFECTIVE AND SAFE METHOD FOR REMOVAL OF COMMON BILE
DUCT LARGE STONES: DATA FROM A REFERRAL CENTER
Di Mitri R.*, Mocciaro F., Pecoraro G.M.
Gastroenterology and Endoscopy Unit, ARNAS Civico-Di Cristina-
Benfratelli Hospital, Palermo, Italy
Background and aim:
Large stones of common bile duct (CBD) are
a hard challenge after endoscopic sphincterotomy (EST) especially
in those that are unable to be managed with standard techniques
including mechanical lithotripsy. Dilation-assisted stone extraction
(DASE) after EST can be more efficient than EST alone for removal of
large CBD stones.
The aim of this study is to report the experience of a referral centre
on the efficacy and complications of DASE for CBD large stones
treatment.
Material and methods:
From January 2013 to September 2015 data
of all consecutive patients who underwent DASE due to large stones,
evidenced by CT-scan or MRI, were collected and recorded in an
electronic database for the final analysis. After selective cannulation
of the CBD, an initial cholangiogram was taken before balloon
placement. The size of the balloon was matched to the diameters
of the bile duct and stones. The balloon was gradually filled with
diluted contrast medium under endoscopic and fluoroscopic
guidance to observe the gradual disappearance of the waist in the
balloon, which was taken to indicate progressive dilation of the