P013 The effects of Schistosoma japonicum eggs antigen on

Basic Science
high dose VitD3 were signiﬁcantly decreased (P < 0.05); the
activity of caspase-3, TNF-a, CD40 in all drug intervention
groups were signiﬁcantly decreased (P < 0.01); no signiﬁcant
difference in mRNA expression was found between TNBS
group and all drug intervention groups. Rats in continuous
high dose VitD3 group had hyperealcemia and serum level
of creatinine was signiﬁcantly increased in continuous high
dose VitD3 group when compared with the other groups
(P < 0.05).
Conclusions: VitD3 can alleviate TNBS induced experimental
colitis by regulating T cell immunity.
P011
The impact of omentectomy on the severity of colitis and
adipocytokine expression
S. Yapali1 *, N. Oruc1 , C. Yenisey2 , M. Sezak3,4 , N.G. Ceri5 ,
O. Ozutemiz1 . 1 Ege University, School of Medicine,
Gastroenterology, Izmir, Turkey, 2 Adnan Menderes
University, School of Medicine, Biochemistry, Aydin, Turkey,
3
Ege Universit, School of Medicine, Pathology, Izmir, Turkey,
4
Ege University, School of Medicine, Pathology, Izmir, Turkey,
5
Adnan Menderes University, School of Medicine, Anatomy,
Aydin, Turkey
Background: Regarding the unidentiﬁed role of omental
adipose tissue on intestinal inﬂammation, we hypothesized
that complete removal of omentum may alter the severity
of intestinal inﬂammation by changing cytokine and adipokine
expression. We aimed to determine whether removal of
omentum has any impact on the severity of colitis and the
expression of cytokines and adipokines in an experimental
model of trinitrobenzene sulphonic acid (TNBS) colitis induced
after 6 months following omentectomy.
Methods: Forty Wistar albino rats (20 M, 20 F) were divided into
two groups. Group A (n = 20) underwent laparotomy and Group B
(n = 20) underwent sham laparatomy without omentectomy.
After 6 months following omentectomy, induction of colitis
was performed by a single intracolonic administration of
0.25ml of 30% ethanol (V/V) containing 30 mg of 2,4,6TNBS. Control groups received 0.25 ml of vehicle (30%
ethanol) with a similar method. Rats were divided into
four groups: 1. Omentectomy & TNBS colitis; 2. Sham
operated & TNBS colitis; 3. Omentectomy & ethanol; 4. Sham
operated & ethanol. Distal colon and mesenteric fat tissue
were removed for histological and biochemical assessments.
Myeloperoxidase (MPO) activity, tumor necrosis factor-alpha
(TNF-alpha), interleukin (IL)-18, were determined in colonic
tissue homogenates by enzyme-linked immune sorbent assay
(ELISA). Leptin and adiponectin levels were determined both
in colonic and mesenteric fat tissue homogenates by ELISA.
Expression of adipocytokines and severity of colitis were
compared between the groups.
Results: Macroscopic and histologic scoring of colitis tended
to be higher in rats who underwent omentectomy and
administered TNBS, the severity of colitis and MPO levels
were not signiﬁcantly altered with omentectomy. TNF-alpha
(p = 0.006), IL-18 (p = 0.002) and adiponectin (p = 0.004) levels
were signiﬁcantly higher in Group 1 than those in Group 2.
Omentectomy did not alter colonic leptin levels signiﬁcantly.
Moreover, mesenteric adipocytokine levels did not change
signiﬁcantly between the groups.
Conclusions: Omentectomy led to signiﬁcantly increased
TNF-alpha, IL-18 and adiponectin levels that was accompanied
by an increased tendency of colonic damage. The effects of
increased cytokines on microscopic and histological ﬁndings
may be a subsequent event. Removal of omental adipose tissue
might also enhance dysregulation of mesenteric adipocytokine
expression. This is the ﬁrst study investigating long-term effects
of omentectomy in an experimental model of colitis and
S69
demonstrating that omentum keeps the novel protective role
in intestinal inﬂammation.
P012
The effects of mesenchymal stem cells as a cell therapy
in animal model of Crohn’s disease
B.I. Jang1 *, W.Y. Jung2 , I.H. Song2 , K.O. Kim1 , C.H. Yang3 ,
G.A. Song4 . 1 Yeungnam University College of Medicine,
Division of Gastroenterology and Hepatology, Department of
Internal Medicine, Daegu, Korea, Republic of, 2 Yeungnam
University College of Medicine, Department of Anatomy,
Daegu, Korea, Republic of, 3 Dongguk University College
of Medicine, Division of gastroenterology and Hepatology,
department of Internal Medicine, Kyungju, Korea, Republic
of, 4 Pusan National University School of Medicine, and
Biomedical Research Institute, Division of Gastroenterology
and Hepatology, Department of Internal Medicine, Busan,
Korea, Republic of
Background: Inﬂammatory bowel disease (IBD) is a complex
immunological disease characterized by chronic inﬂammation
with unknown cause. Interleukin-10 Knockout (IL-10 KO) mouse
is a well-established murine model of IBD which develop a
spontaneous intestinal inﬂammation that resemble with Crohn’s
disease.
In the present study, human adipocyte derived stem cells
(hAMSC) were administrated to IL-10 KO mice to ﬁnd out antiinﬂammatory effects of hAMSCs may attenuate progress or treat
of IBD.
Methods: IBD was induced in IL-10 KO mouse by feed piroxicam
mixed diet for 1 week and then, 2×106 hAMSC were injected
into peritoneum and changed to normal diet. After 1 week,
the mice were sacriﬁced and tissue sample was harvested and
tissue score for inﬂammation and inﬂammation related gene
expression were evaluated.
Results: hAMSC administrated group showed reduced inﬂammatory changes signiﬁcantly. The effect was more powerful
in ascending colon than descending or cecum in regional
analysis. In categorical analysis, protection effect on chronic
inﬂammation was more evident than acute inﬂammation
considering ordinary progress of IBD; inﬂammatory cells appear,
goblet cells change, mucosa thickening, submucosa cell
inﬁltration, and destruction of architecture. RT-PCR analysis
showed that Rantes, toll like receptor, IL-4 expression were
not noticeably different between groups but IL-12, INF, TNF-a
showed signiﬁcant decrease in hAMSC group.
Conclusions: hAMSC attenuate IBD induction in IL-10 KO
mouse by suppression of inﬂammatory cytokine expression
and this expected to be inﬂuenced through type 1 helper
T cell pathway. Even though single injection of hAMSC, the
effect inﬂuenced chronic event of inﬂammatory changes.
These ﬁndings suggested that administration of hAMSCs show
beneﬁcial effect on animal model of CD and emerge a
therapeutic option in inﬂammatory bowel disease.
P013
The effects of Schistosoma japonicum eggs antigen on
SAMP1/Yit mice, a spontaneous ileitis model
T. Arai *, T. Seki, T. Kumagai, R. Shimogawara, N. Ohta.
Graduate School of Tokyo Medical and Dental University,
Department of Environmental Parasitology, Bunkyo-ku,
Japan
Background: It is known that the development of Crohn’s
disease (CD) is related to a polarized Th1 response.
Exposure to Schistosoma japonicum (SJ) eggs causes strong
Th2 response, which is inhibitory against the Th1 response.
Furthermore, it activates a network of regulatory T cells (Tregs)
that secret interleukin-10. This cytokine down-regulate Th1
response in mice. Several studies showed that exposure to
SJ eggs protected mice from experimental colitis. However,
S70
these experiments have been conducted with drug induced
models, such as 2, 4, 6-trinitrobenzene sulfonic acid (TNBS).
Considering that drug induced models are artiﬁcial and
affect large bowel rather than small intestine, it is not
suitable for studies on CD. A senescence-accelerated mouse
(SAM) P1/Yit is a strain that develop spontaneous ileitis
with 100% penetrance until 30 weeks of age, while losing
its senescence accelerated phenotype. In this study, we
investigated that whether exposure to SJ eggs prevent
spontaneous ileitis.
Methods: Speciﬁc pathogen-free SAMP1/Yit mice were donated
by the Yakult Central Institute for Microbiological Research
(Tokyo, Japan). AKR/NSLc mice were used as control mice.
SJ eggs were collected from the liver of Schistosome-infected
mice. The eggs were washed in phosphate buffered saline
(PBS). Mice were injected with freeze-killed 5,000 eggs
intraperitoneally once a week from 12 to 29 weeks of age.
Control mice received PBS with the same volume. The effects
of SJ eggs were quantiﬁed as follows: 1) body weight loss,
2) histological score of microscopic inﬂammation of ileum,
3) cytokines proﬁles of T-lymphocytes which are isolated
from mesenteric lymph nodes (MLN). Statistical analyses were
performed using Student’s t test. P < 0.05 was considered to be
statistically signiﬁcant.
Results: 1) SAMP1/Yit mice treatment with SJ eggs showed
tendency to gain weight. 2) Histological score of ileitis was
not modulated. 3) In the treatment group, the concentration
of Interleukin (IL) 4 was increased signiﬁcantly, however
interferon (IFN)-gamma and tumor necrosis factor (TNF)-alpha
were not decreased and IL-10 was not increased compared to
the control group.
Conclusions: This is the ﬁrst report that investigated the
effects of SJ eggs antigen on spontaneous ileitis model of
CD. These ﬁndings suggest that treatment with SJ eggs did
not attenuate ileitis, but increased body weight and the upregulation of IL-4 suggest the therapeutic potentials of the
parasite antigen for CD.
P014
The effect of the anti-TNFa adalimumab on the levels of
angiogenic factors in mucosal cultures from patients with
inﬂammatory bowel disease (IBD)
andez-Contreras1 , A. Algaba2 ,
P.M. Linares1 *, M.E. Fern´
M. Chaparro1 , F. Bermejo2 , J.P. Gisbert1 . 1 Hospital
Universitario de La Princesa, IP and CIBERehd,
Gastroenterology Unit, Madrid, Spain, 2 Hospital Universitario
de Fuenlabrada, Gastroenterology Unit, Fuenlabrada, Spain
Background: Effectiveness of adalimumab (ADA) treatment
could be related to the modiﬁcation of different angiogenic
proteins, including VEGFA, Ang1, Ang2 and their receptor Tie2.
The aim of our study is to compare the concentrations of several
angiogenic factors in colonic mucosa culture supernatant (MCS)
in patients with IBD and matched controls and to analyze
their modiﬁcations with the in vitro ADA addition to the
culture.
Methods: Prospective study in patients with IBD and nonIBD controls that underwent endoscopy. Duplicates of colonic
mucosa samples from affected and non-affected mucosa
from each IBD patient were obtained for comparison. Both
were washed and then cultured at 37ºC in 5% CO2 medium
under shaking for 24hours. In one duplicate, ADA was added
up to a ﬁnal concentration of 10 mg/mL prior to culture.
MCS levels in AP were determined by ELISA. Endoscopic
ulcerative colitis (UC) and Crohn’s disease (CD) activity
was ascertained by Mayo sub-score and SES-CD indexes,
respectively.
Results: 28 patients with IBD (16 UC, 12 CD) and 21 controls
were included. Mean age was 41±16 years, and 61% were
women. The mean disease duration was 7±7 years. According
Poster presentations
to endoscopic activity, 36% of patient had quiescent, 32% mild,
28% moderate, and 4% severe activity. All angiogenic factors
mean levels in MCS were higher in affected than in non-affected
MCS; VEGFA (19.1±18.7 versus 8.7±10.5 pg/mL/mg) and Ang2
(21.3±15.7 versus 11.8±8.9 pg/mL/mg) (p < 0.05). There were
no differences in MCS depending on endoscopic activity. Levels
of VEGFA, Ang1 and Ang2 from the affected, and VEGFA and
Tie2 from the non-affected mucosa, were lower when cultured
with ADA than without it (Table 1).
Table 1. Levels of angiogenic factors in MCS from IBD patients
after culture
VEGFA
Ang1
Ang2
Tie2
Affected
mucosa
Affected
mucosa
+ ADA
p-value Nonaffected
mucosa
Nonaffected
mucosa
+ ADA
p-value
19.1±18.7
15.7±8.8
21.3±15.7
12.0±5.5
14.6±14.5
12.2±6.2
19.4±13.1
12.4±16.1
0.003
0.021
0.021
0.911
5.8±6.1
9.1±7.1
10.4±6.7
5.3±5.2
0.027
0.069
0.406
0.021
8.6±10.5
12.5±9.3
11.8±8.9
8.8±7.0
Concentrations are expressed in pg/mL per mg of tissue.
In UC patients, VEGFA and Ang1, and all angiogenic factors
MCS mean levels were lower in the affected-mucosa and
non-affected mucosa, and in the non-affected mucosa when
cultured with ADA, respectively (Table 2).
Table 2. Levels of angiogenic factors in MCS from CD patients
after culture
VEGFA
Ang1
Ang2
Tie2
Affected
mucosa
Affected
mucosa
+ ADA
p-value Nonaffected
mucosa
Nonaffected
mucosa
+ ADA
p-value
20.4±13.9
18.2±9.6
27.2±14.6
12.7±4.6
15.7±10.9
13.8±6.4
25.7±10.9
14.7±18.8
0.015
0.049
0.743
0.720
5.0±6.0
8.7±5.8
12.2±7.8
4.4±3.1
0.015
0.020
0.001
0.026
7.0±7.2
13.0±9.6
14.5±9.8
7.5±3.7
Concentrations are expressed in pg/mL per mg of tissue.
In contrast, in CD patients there were no differences in MCS
angiogenic factors levels and the addition of ADA to the
culture.
Conclusions: ADA might downregulate the production of
angiogenic factors in MCS. ADA addition to mucosal cultures
affects angiogenic factors levels in samples from patients with
UC differently than in patients from CD.
P015
The VEGFC/VEGFR3 pathway controls colitis associated
cancer formation and progression through
lymphangiogenesis and lymphatic endothelial VE-cadherin
C. Tacconi *, S. D’Alessio, C. Correale, A. Gandelli, S. Vetrano,
S. Danese. Humanitas Research Hospital, IBD Center, Rozzano,
Milan, Italy
Background: Chronic intestinal inﬂammation occurring in
Inﬂammatory Bowel Disease (IBD) predisposes to colitisassociated colorectal cancer (CAC). Tumor-induced lymphangiogenesis is one of the processes taking place in cancer,
but no mechanistic data is available on the molecules
modulating lymphangiogenesis in CAC. VEGFC is a key player
in lymphangiogenesis that signals through its cognate receptor
VEGFR3 and its blockade has been used in several models of
metastatic cancer. However, little is known about the role
played by adherens’ junction proteins, such as Vascular Endothelial (VE)-cadherin, on the lymphatic endothelium during
the metastatic process. We hypothesized that lymphaticstargeted therapy may be effective in CAC formation and would

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