Curcumin alleviates experimental colitis via a potential mechanism

involving memory B cells and Bcl-6-Syk-BLNK signaling

Si-Yi Wei, Tian-Tian Wu, Jia-Qi Huang, Zeng-Ping Kang, Meng-Xue Wang, You-Bao Zhong, Wei Ge, Bu-Gao Zhou, Hai-
Mei Zhao, Hai-Yan Wang, Duan-Yong Liu

Rohani
260120220512

Dosen pengampu : Dr. Med. apt. Melisa Intan Barliana

Universitas Padjadjaran
Background
1. As a chronic nonspecific
inflammatory disorder,
inflammatory bowel disease
(IBD), classified into 3. Curcuma longa L, also named
ulcerative colitis (UC) and turmeric have bioactive natural
Crohn’s disease (CD), is polyphenols called curcumin (Cur)
characterized by recurrent [1,7-bis(4-hydroxy-3-
abdominal pain, diarrhea, methoxyphenyl)- 1,6-heptadiene-
anemia, bleeding, and weight 3,5-dione].
loss.

4. As an immune suppressant,
2. Immune system curcumin (Cur) has a therapeutic
dysfunction has been regard role in treating many immune
as the main cause of IBD, diseases, such as IBD and
which is mainly involving rheumatoid arthritis.
abnormal lymphocytes
(including T and B cells,
including memory B cells),
macrophages, mast cells, and
activated neutrophils
Aim
To investigate whether or not curcumin (Cur) can alleviate experimental colitis by regulating
memory B cells and Bcl-6-Syk-BLNK signaling
Materials and Method
A. Animals

BALB/c male mice, aged 8-9 wk and weighing 20-

22 g

Control group, in which DSS + Cur group, in

Control + Cur group, in DSS group, in which
mice were fed normal which mice with DSS-
which normal mice were mice with DSS-induced
food without DSS or drug induced colitis were
given Cur treatment colitis
treatment treated with Cur.
B. DSS-induced experimental colitis

Mice were freely given a 3% (w/v) solution of DSS) in drinking water for 7 d, then
with sterile drinking water for 7 d, and finally with 2% (w/v) DSS for 7 d

Cur was dissolved in 1.5% sodium carboxymethylcellulose, mice in the DSS + Cur
group and Control + Cur group were administrated with Cur (100 mg/kg/d) by
gavage for 14 consecutive days

Mice in the Control and DSS groups were administrated with an equal volume of
saline starting from day 8
C. Histological evaluation

All mice were weighed before anesthesia with sodium pentobarbital (20 mg/kg i.p.)

the whole colons were rapidly separated on the ice box, and their length and weight
were measured

the colonic pathological changes were observed under a microscope 

D. Flow cytometry
500 μL of peripheral blood from each mouse was collected and treated with 1 mL of
lysing buffer and incubated for 15 min in the darkness to remove red blood cells

The cell suspension was combined with an Fcγ receptor-blocking mAb for 15 min
at 4 °C

The cells were detected for surface antigens by labelling with BV510 rat anti-
mouse CD19 , AF488 rat anti-mouse CD27 , PE-A rat anti-mouse IgM ,BV421 rat
anti-mouse IgG , APC rat anti-mouse IL-10, BV421 rat anti-mouse IgA , APC rat
anti-mouse FCRL5, PE-A rat anti-mouse CD103 , BV421 rat anti-mouse FasL ,
APC rat anti-mouse PD-1 , BV421 rat anti-mouse CXCR3 , and PE-A rat anti-
mouse Tim-3

The single-cell suspensions were analyzed with a FACS Canto II flow cytometer
E. ELISA
One hundred micrograms of mouse colon tissue was homogenized by adding 1000
μL of RIPA lysis buffer

The samples were incubated at 4 °C for 1 h, ultrasonically homogenized, and

centrifuged at 13000 rpm for 10 min

ELISA kits were used to detect the secretion of IL-1β, IL-6, IL-10, IL-35, IL-17A,
and TNF-α
D. Western blot analysis

Colonic mucosa protein were resolved by polyacrylamid gel electrophoresis and

transferred onto PVDF membranes, which were then blocked with 3% bovine
serum albumin (BSA) solution for 1 h at room temperature and then incubated with
the following primary antibodies overnight at 4 °C; BLNK, p-BLNK, Syk, p-Syk,
CIN85, Bcl-6, and anti-GAPDH antibodies.

HRP-coupled secondary antibody was added and incubated for 2 h at room

temperature

ECL hypersensitive solution was used to visualize the protein blotting. 

Photographs were taken by using the Highly Sensitive Chemiluminescence Imaging

system
Result
A. Cur alleviates DSS-induced colitis
B. Cur regulates inflammatory cytokine expression
C. Cur regulates memory B cell subsets
D. Cur regulates the activation of the Bcl-6-Syk-BLNK signaling pathway
Conclusion
Cur could effectively alleviate DSS-induced colitis in mice by regulating memory B cells and the Bcl-6-Syk-
BLNK signaling pathway.
Reference
Wei SY, Wu TT, Huang JQ, Kang ZP, Wang MX, Zhong YB, Ge W, Zhou BG, Zhao HM, Wang HY, Liu DY.
Curcumin alleviates experimental colitis via a potential mechanism involving memory B cells and Bcl-6-
Syk-BLNK signaling. World J Gastroenterol. 2022 Oct 28;28(40):5865-5880. doi:
10.3748/wjg.v28.i40.5865. PMID: 36353208; PMCID: PMC9639655.
Thank you!