molecular biology article

1. Valentina Dávila Arenas
María Daniela De la Ossa Posada
Third semester
School of Medicine UPB
Published: 21 january of 2021

2. OBESITY
What is it?
Obesity is a chronic, complex and
multifactorial disease.

3. ADH1B
What is it and for what do we use it?
*ADH is an enzyme that catalyzes many reactions, in
this study the ADH1B subunit was used to see an
inverse correlation with obesity and IR

4. “We performed molecular studies, using human subcutaneous
pre-adipocyte cell lines to examine the role of ADH1B protein in
adipocytes and the potential involvement of ADH1B in the
development of obesity.”
GOAL OF THE ARTICLE
HUMAN PRE-ADIPOCYTES

5. METHODS
Fundament what do we use it for?
Western blot analysis.
It’s used to investigate the potential function
of ADH1B protein in adipose tissue. They
examined in vitro protein expression levels
of ADH1B in differentiated subcutaneous
human pre-adipocytes derived from adipose
tissue.
The method involves the use of gel electrophoresis to
separate the proteins from the sample. The separated
proteins are transferred from the gel to the surface of
a membrane. The membrane is exposed to a specific
antibody against the protein under study. Antibody
binding is detected using a radioactive or chemical
marker

6. METHODS
Fundament what do we use it for?
Glucose uptake assay
The glucose uptake assay protocol is
based on a fluorescent glucose analog,
which just like glucose can be taken up by
cells through glucose transporters. This
glucose analog cannot be fully utilized in
glycolysis because of its modification and
thus accumulates inside the cells.
Fluorescence generated by the fluorescent
glucose analog is proportional to the
glucose uptake by the cells
It is used to measure glucose
uptake using fluorescent
microscopy and flow cytometry.
In the studio, they comprobate that
knockdown of ADH1B expression
decreased insulin-stimulated
glucose uptake
The results showed that deficiency in ADH1B
protein expression significantly reduced
adipocyte glucose uptake following insulin
treatment by almost 40% compared to control

7. METHODS
RNA interference (RNAi).
Fundament
what do we use it
for?
Is a biological process in
which RNA molecules
inhibit gene expression or
translation by neutralizing
targeted mRNA molecules.
They used it to evidence
that knockdown of
ADH1B yields a decrease
in insulin-stimulated
glucose uptake within
adipocytes.

9. METHODS
Fundament what do we use it
for?
Quantitative polymerase chain reaction
(qPCR)
It’s an enzymatic DNA
amplification
technique where
polymerase starts the
replication simulation
Q: quantification
It is used to
identify genes
and relate their
expression in
obesity

10. RESULTS
The expression of ADH1B only starts to
show from day 2 and is directly
proportional to the days, the expression
of B actin is always present because it is
a protein that is used in all cell lines as
a test control.
ADH1A
ADH1C
They are not expressed because they
have a higher concentration in the liver
than in the adipose tissue.

11. RESULTS
Proteins and genetic
material were evaluated in
this graph you can see the
increased expression in
ADH1B in myc.
FABP4 has a decreased
expression

12. RESULTS
In this result, we can saw that Insulin
promotes expression of ADH1B in lean
and obese adipocytes.
*at a higher insulin concentration, we have a
higher expression of ADH1B.
*B-actin is the control protein.

13. RESULTS
*ADH1B expression with ( +) and without (−) treatment with AKT inhibitor and/or insulin.
*Non-treated cells (−Insulin, −AKT inhibitor) were utilized as control.
*β-Actin was used as loading control.
Mature adipocytes were treated with an
AKT-specific inhibitor to suppress phosphorylation
of AKT to the active form.
Western blot analysis of expression levels of total
AKT protein (AKT), phosphorylated AKT (p-AKT),
and ADH1B

14. DISCUSSION
Author What does he said? Agree or not
Winnier,D A.(22)
Our previous and current
findings have also revealed that
ADH1B gene and protein
expression levels are clearly,
inversely correlated with BMI
DeFronzo, R. A(4)
DeFronzo, R. A. & Prato, S.
D.(6)
We and others have demonstrated
that T2D is an inherited disease
characterized by insulin
resistance in insulin target
tissues, notably adipose tissue,
skeletal muscle, liver and
pancreatic b-cells
Burke, J. P(11)
Duggirala, R(13)
We previously showed that
obesity, insulin resistance and
T2D traits have a strong genetic
basis and high prevalence rates
in Mexican Americans

15. CONCLUSIONS
1. The expression of the protein and the ADH1B gene is inversely proportional to
the BMI, it means that the more the BMI increases, the lower the expression of
this gene will be.
2. ADH1B plays a functional role in the cell response to insulin: insulin induced
ADH1B expression results in a proportional increase in activity. And also
induced enzyme activity.
3. Knockdown of ADH1B expression decreased insulin-stimulated glucose uptake.
Also a higher insulin concentration shows a higher expression of ADH1B and
this increased expression is related to a low BMI.