The study used proteomic analysis to examine the aortas of human lipoprotein(a) transgenic mice. The analysis found an early metabolic response in the transgenic mice that was independent of atherosclerosis. Specifically, it showed changes in arterial protein levels compared to wild type mice. The goal was to determine proteomic changes before atherosclerotic lesions appear to better understand disease development at the early stage. The results provide insights into molecular mechanisms influenced by elevated lipoprotein(a) levels independently of atherosclerosis.
1. PROTEOMIC ANALYSIS OF AORTAE FROM
HUMAN LIPOPROTEIN(A)
TRANSGENIC MICE SHOWS AN EARLY
METABOLIC RESPONSE
INDEPENDENT OF ATHEROSCLEROSIS
Euan J. Rodger, Rachel J. Suetani, Gregory T. Jones, Torsten Kleffmann, Alan Carne,
Michael Legge,Sally P. A. McCormick.
Esteban Vallejo Agudelo
María Camila Vélez Muñoz
TERCER SEMESTRE
2. INTRODUCTION
Many transgenic mice
models were used in
studies of atherosclerosis
The apoE-
deficient mouse is the
most spontaneous lesions t
o form assembled with
human-like
This mouse has been used
in genetic and drugs
studies to
identify molecules that
modify the disease
development
3. INTRODUCTION
Studies in the early stage of
atherosclerosis before lesions
appear were very few.
Quantitative proteomics studies can detect
changes in cellular proteins and reveal
molecular mechanisms of disease
4. INTRODUCTION
Proteomic changes in apoE
deficient mice at different stages of
the lesion showed an
imbalance and a deterioration
of energy
metabolism probably given by
increased levels
of remnant lipoproteins
5. INTRODUCTION
Humans have a
lipoprotein that is not in
mice, the lipoprotein has
an LDL coupled with
a apoprotein(a).
6. INTRODUCTION
Some studies
Other studies
found that Lp (a) have
found that low levels of
a moderate
this may decrease the
risk associated with
risk of coronary heart
the development of
disease.
atherosclerosis.
These studies show that the
combination of elevated levels
of LDL and Lp (a) with a high fat
diet may be atherogenic.
7. CONCEPTS
PROTEOMIC
Is the study of all proteins in a
genome allowing to classify
them according to
their function and
interactions between them.
8. CONCEPTS
LIPOPROTEINS
Lipoprotein(a)
Molecules composed of protein conteins Apo(a)
and fat that have the function to
transport cholesterol and other
substances through the blood.
Interferes
with fibrinolysis and is
a risk factor for developing
diseases
9. CONCEPTS
Atherosclerosis : Is the
hardening of the arteries, presented
for the accumulation of cholesterol
and fats along the walls of the arteries.
This plate prevents the passage of
blood and oxygen to the tissues.
Transgenics: Are beings that has
been made through genetic
manipulation.
10. RELATIONSHIP
ATHEROSCLEROSIS LIPOPROTEINS
TRANSGENICS PROTEOMICS
Lp(a)
The relationship between them is that in
this study used proteomic analysis of the
arteries in mice
with transgenic manipulation genetics for
finding high levels of Lp(a)
12. MATERIALES Y METODOS
RATONES
Ratones de tipo salvaje C57BL/6
Se utilizaron 40 ratones: 20
Transgénicos y 20 de tipo Salvaje
Edad: 30 semanas
13. MATERIALES Y METODOS
CORTES HISTOLOGICOS
1. 8 ratones de tipo transgénico y 8 de tipo salvaje.
2. Fijación en formol y otras sustancias.
3. Coloración con hematoxilina y eosina, más otro como el Verhoeff.
4. Anticuerpos de conejo.
14. MATERIALES Y METODOS
ANALISIS DE LIPIDOS EN EL PLASMA
Los niveles Los niveles plasmáticos
plasmáticos de de Lp(a) y apoB se
colesterol, triglicéridos midieron utilizando las
y fosfolipidos se enzimas especificas de
midieron utilizando apo(a) y apoB en el
reactivos enzimáticos método ELISA.
15. MATERIALES Y METODOS
ANALISIS DE LIPIDOS EN EL PLASMA
La distribución de los
lípidos en el plasma se
analizó por la separación
de las lipoproteínas en el
plasma.
16. MATERIALES Y METODOS
ANALISIS DE LIPIDOS EN LA AORTA
Los lípidos se
extrajeron mediante
cloroformo y
metanol
Se resuspendieron
mediante
isopropanol
17. MATERIALES Y METODOS
ANALISIS DE LIPIDOS EN LA AORTA
Los niveles de
El acido tiobarbiturico
colesterol, triglicéridos
se utilizo para medir la
y fosfolipidos se
concentración de
midieron mediante
aldehídos para análisis
enzimas de
de lípidos oxidados.
restricción.
18. MATERIALES Y METODOS
2D PAGE – ELECTROFORESIS BIDIMENCIONAL
En gel de poliacrilamida.
Se utiliza para el análisis global
de la separación de los
componentes del proteoma.
Separación de muchas
proteínas en un único gel.
19. MATERIALES Y METODOS
RT-PCR CUANTITATIVA
Se usa para amplificar y
En la investigación se
cuantificar el cDNA y así
utilizo para cuantificar el
tener datos precisos
nivel de transcripción del
sobre la expresión génica
RNAm en las proteínas
de las células en el
de interés en la muestra.
estudio.
20. RESULTADOS
Uso de Anticuerpo
específico de ApoB humana
ApoB libre mucho mayor a
unida.
21. RESULTADOS
Uso de Anticuerpo
específico de Apo(a)
humana
Toda la apo(a) estaba unida
a ApoB en forma de Lp(a)
27. DISCUSSION
RESEARCHERS RESULTS RELATIONSHIP
Mayr M, Chung YL, Impaired fatty, energy Agree, but the proteins
Mayr U, Yin X, Ly L, et and reduction found were different.
al. metabolism but without
vascular damage
Makowski L, Boord JB, Decrease in FABP4 Not agree, because
Maeda K, Babaev VR, levels deficient levels of
Uysal KT, et al FABP4 are protective
factors against
atherosclerosis
28. DISCUSSION
RESEARCHERS RESULTS RELATIONSHIP
Yvan-Charvet L, Wang Increased plasmatic Agree, might be a early
N, Tall AR levels of HDL. protective factor
against atherosclerosis
Feil S, Hofmann F, Feil Decrease of Not agree, that´s
R cytoskeletal proteins contradictory when
(trangeline y Mfap4) atherosclerosis is
beginning
29. CONCLUSIONS
• The interaction with the environmental factors affect
the expression of our genetic information, so is possible
to find a change in protein production when there is an
increase in fatty acids.
• The level of different lipoproteins contributes directly or
indirectly in the formation of vascular lesions that can
become atherosclerosis and
cause serious cardiovascular disease.
30. CONCLUSIONS
• Atherosclerosis is a condition that is influenced
by lifestyle and diet, but has a strong genetic basis.
• Atherosclerosis is a disease of high incidence
with high levels of mortality. There have been
many studies but still found conflicting results
and unrelated data. It is therefore very important to
continue studying further.
