Referencias: - Dextran Sulfate Sodium (DSS)‐Induced Colitis in Mice - Flow Cytometry Fundamental Principle https://www.bosterbio.com/protocol-and-troubleshooting/flow-cytometry-principle - Hayden MS, Ghosh S. Signaling to NF-kappaB. Genes Dev. 2004;18:2195-2224. - http://www.scielo.org.co/scielo.php?script=sci_arttext&pid=S0120-00112008000200006

1. Carla Vega Salazar
Medicine
Third Semester
2020-01

2. I N T R O D U C T I O N
COLITIS
Inflammatory bowel disease
(IBD)
https://www.manhattangastroenterology.com/conditions/ulcerative-colitis/
• Genetic susceptibility.
• Intestinal microflora.
• Inmune factors.
• Fever.
• Joint pain.
• Rectal bleeding/Diarrhea.
• Weight loss.
Anti-inflammatory and
immune-regulatory
medications.

3. I N T R O D U C T I O N
DSS
Chemical
colitogen with
anti-coagulant
properties.
GOSS
Anti-inflammator
y
Anti-aging
Anti-cancer
BIFIDUS FACTOR
NF-kB
Protein complex
that controls
DNA
transcription
Oxidative and genotoxic
stress -> DNA damage
pathway.

4. O B J E C T I V E
Explore the effects of Galactooligosaccharides (GOS) on
intestinal flora and its anti-inflammatory effects in Dextran
Sulfate Sodium (DSS) induced murine colitis and
explorate the underlying mechanism.

5. M E T H O D S
Sample containing cells
marked with fluorescent
substances, absorb a laser
beam that causes them to
emit a characteristic
wavelength of the cell
population that is going to be
measured.
Measure the percentage
of Treg and Th17 cells.
F L O W C Y T O M E T R Y
F U N D A M E N T W H A T F O R ?
https://www.bosterbio.com/protocol-and-troubleshooting/flow-cytometry-principle

6. Use of an enzyme to detect
the presence of a ligand in a
liquid sample, using
antibodies directed against
the protein that is going to be
measured.
Quantification of
cytokines secretion and
mRNA expression
M E T H O D S
E N Z Y M E L I N K E D I M M O N O A B S O R B E N T A S S A Y (ELISA)
F U N D A M E N T W H A T F O R ?

7. Thermocycler induces a
beam of light of an
established wavelength and
detects fluorescence by the
excited fluorochrome of the
sample cells
Quantification of
cytokines secretion and
mRNA expression
M E T H O D S
R E A L T I M E P C R (RT-PCR)
F U N D A M E N T W H A T F O R ?

8. Identify the presence of a
specific protein in
electrophoretic separated
samples using
immunoblotting (specific
antibodies)
Measure the percentage of
cytosolic and nuclear
proteins on the colon tissue
M E T H O D S
W E S T E R N B L O T
F U N D A M E N T W H A T F O R ?

9. R E S U L T S
1d: Intragastrical GOS (2-4) and regular
water (1-3).
3d: Tracking day.
5d: Tracking day.
6d: Tracking day.
7d: Mice were sacrificed.
GROUPS
1: Negative control group.
2: GOS group.
3: DSS model group.
4: DSS + GOS treatment group.

10. R E S U L T S

11. R E S U L T S

12. R E S U L T S

13. D I S C U S S I O N
AUTHOR HYPOTHESIS YES/NO
Wendy S. Garrett, Jeffrey
I. Gordon, Laurie H. Glimcher
“Both innate and adaptive immunity have been suggested
playing a key role in maintaining intestinal homeostasis and
there are various factors involved in this process. A breakdown
in this well-controlled balance could result in intestinal
disorders, such as ulcerative colitis and increased infectious
diseases”
Amrita Kumar, Huixia Wu,
Lauren S Collier, Hyams
Jason M, Hansen Tengguo Li
Kosj Yamoah, et al.
“Inflammation is usually initiated when host cells come in
contact with microbes and is dependent on the activation of
nuclear factor κB (NF-κB) signaling pathway”
Atreya, R. Atreya, MF
Neurath
“Inhibition of NF-kB activation has been suggested as an
anti-inflammatory strategy in UC”

14. C O N C L U S I O N S
GOS have positive effects in the treatment of
DSS-induced colitis, which gives a possible
effective treatment for ulcerative colitis in humans.
The implementation of anti-inflammatory
molecules such as GOS, could benefit the
treatment of other related diseases.

15. C O N C E P T U A L M A P