UEG Journal Abstracts 2022 - 96
The microbiome as disease modulator
11:00-12:00 / Lehar 4
OP121
PEPTOSTREPTOCOCCUS ANAEROBIUS PROMOTES GASTRIC MUCOSAL
LESIONS THROUGH TLR4/TRIF/NF-ΚB SIGNALING PATHWAY
Y. Ouyang1
1
, N. Li1
, C. Peng1
, C. He1
, C. Xie1
, Y. Hu1
, Z. Yin1
The First Affiliated Hospital of Nanchang University,
Gastroenterology, Nanchang, China
Contact E-Mail Address: 18720997548@163.com
Introduction: In addition to Hp, there are a large number of bacteria in the
stomach to participate in gastric mucosal lesions. Our study found that
compared with patients with gastritis, the abundance of gastric mucosal
Peptostreptococcus anaerobius (Pa) in patients with gastric cancer was
significantly increased, indicating that it may be involved in the process of
gastric mucosal carcinogenesis.
Aims & Methods: Human samples: The bacterial flora of gastric mucosa
was detected by 16SrDNA sequencing, and the significant difference between
gastritis and gastric cancer was detected by LEFSe analysis and
FISH experiments.
Cell model: The AGS co-cultured with the AGS cell model was constructed,
and biological behavior experiments were performed. Cell total mRNA
was extracted and sequencing was performed. The mRNA and protein expression
of key proteins of NF-κB and TLRs, TRIF, Myd88 was detected by
q-PCR and WB. The effect of inhibition of TLR4, TRIF on Pa induced NF-κB
activation and the expression of cytokines was also detected.
Animal model: The Pa-infected INS-GAS animal model was constructed.
The feces and gastric mucosa samples of mice were collected to detect the
colonization of Pa. H&E staining and alcian blue nuclear fast red staining
was used to detect the pathology of gastric mucosal sections. The expressions
of CD45 and Ki67 in mouse gastric mucosa were detected by IHC.
qPCR was used to detect the expression of inflammatory factors in mouse
gastric mucosa. WB, IHC, and IF were used to detect the expression of key
proteins of NF-κB and upstream TLR4, TRIF, and Myd88 expression.
Results: Human samples: The abundance of Peptostreptococcus in the
gastric cancer group was significantly higher than that in the gastritis
group. FISH experiment results were consistent with the sequencing results.
The abundance of Pa bacteria in gastric mucosa of patients with gastritis,
intestinal metaplasia, and gastric cancer gradually increased.
Cell model: Pa can cause malignant changes in the biological behavior
of gastric mucosal epithelial cells in vitro. The transcriptome sequencing
results showed that after Pa infected the cells, NF-κB signal pathway
is significantly enriched. Pa bacteria could up-regulate the expression of
p-p65 and p-IKK and downregulate IκBα Expression, promote p65 into the
nucleus.
We found that Pa infection significantly promoted the expression of TLR4,
and TRIF , but not TLR1 and Myd88. The inhibition of TLR4 and TRIF could
inhibit Pa-mediated NF-κB signal pathway activation and cellular inflammatory
response.
Effect of Pa infection on gastric mucosal lesions in INS-GAS mice: H&E
staining of mouse gastric mucosa showed that the gastric mucosa of mice
in the infection group was significantly thickened, some cystic dilated
glands were visible, and obvious inflammatory cell infiltration was visible
at the base of mucosa.
The inflammatory score was significantly higher than that of the control
group. In addition, Pa infection could significantly up-regulate IL-1β,
TNA-α, and IL-18 mRNA expression of mouse gastric mucosa. Pa infection
could up-regulate the expression of TLR4, TRIF, and p-P65 in mouse gastric
mucosa, promote the entry of p65 into the nucleus, and down-regulate
IκBα expression.
96
, N. Lu1
OP122 WITHDRAWN
Conclusion: Pa is a pathogenic bacterium in the stomach. Its infection can
cause gastric mucosal cells and gastric mucosal lesions in ins-gas mice,
especially inflammatory reactions. Pa promotes gastric mucosal lesions
through TLR4/TRIF/NF-κB signaling pathway and inhibiting TLR4 andTRIF
can reduce the pro-inflammatory effect of Pa.
References: NA
Disclosure: Nothing to disclose.
OP123
FACTORS AFFECTING THE OUTCOME OF FECAL MICROBIOTA
TRANSPLANTATION FOR PATIENTS WITH IRRITABLE BOWEL SYNDROME
M. El-Salhy1
1
, T. Hausken2
, J.G. Hatlebakk2
Stord Hospital Medical Clinic Dept. of Gastroenterology/University
of Bergen, Gastroenterology, Stord, Norway, 2
Hospital, Medical Dept, Bergen, Norway
Haukeland University
Contact E-Mail Address: magdy.elsalhy@sklbb.no
Introduction: Fecal microbiota transplantation (FMT) as an intervention
for irritable bowel syndrome (IBS) has been applied in 7 randomized controlled
studies with varied outcome.1
Aims & Methods: In a study of FMT for IBS with a high efficacy, a protocol
for FMTwith a combination of favorable factors wasused.2
The present study aimed at evaluate some of these factors such as the
dose of fecal transplant, the route of administration, and repeated FMT.
The study included 186 IBS patients who randomized 1:1:1 into transplant
administrated to the colon (single LI), to the duodenum (single SI), or to
the distal duodenum twice with a week interval (repeated SI). The fecal
transplant dose was 90g. The patients provided a fecal sample and were
asked to complete five questionnaires to assess their symptoms, fatigue
and quality of life at the baseline, and at 3, 6, and 12 months after FMT.
The fecal bacteria composition and dysbiosis index (DI) were analyzed using16S
rRNA genePCR DNA amplification/probe hybridization covering the
V3-V9 regions.
Results: The response rates in single LI, single SI and repeated SI were
80.4%. 80% and 80.4% after 3 months, 67.9%. 71.4%, and 86% after 6
months, and 51.9%. 75.5% and 80.9% after 12 months.The response rate
was significantly higher in single SI than that of single LI at 12 months after
FMT. The response rates did not differ between single SI and repeated SI at
all observation times after FMT.Symptoms and quality of life improved in
all the treated groups at all time intervals after FMT.
The abdominal symptoms were significantly lower and quality of live
higher in repeated SI group than those of single SI at all observation times
after FMT. DI decreased significantly in all the treated groups at all observation
times after FMT. The bacterial profiles changed in all groups at all
observation intervals.
However, these changes differed between single LI on and single SI/repeated
SI. Of these bacteria 6 were correlated to abdominal symptoms
and 2 were correlated to fatigue. Holdemanella biformisfluorescence signalsincreased
significantly in all groups at 3 and 6 months. Whereas the
Holdemanella biformisfluorescence signals remained significantly higher
than those at the baseline in single SI and repeated SI at 12 months after
FMT, those of the single LI were significantly lower than baseline values.
Furthermore, whereas the Lactobacillusspp. fluorescence signalsincreased
significantly in single SI, and repeated SI at all observation intervals
after FMT, those in single LI increased only at 3 months after FMT.
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