Accelerated fat cell aging links oxidative stress and insulin resistance in adipocytes. Researchers induced oxidative stress in 3T3-L1 adipocytes to study shortened telomeres, senescence, and functional impairment. The stressed adipocytes showed increased reactive oxygen species, DNA damage, and expression of senescence and pro-inflammatory markers. They also exhibited shortened telomere length and decreased glucose uptake. The study aimed to determine how oxidative stress and accelerated senescence impact fat tissue function and how this leads to age-related diseases like obesity and type 2 diabetes.

1. Accelerated fat cell aging
links oxidative stress and
insulin resistance in
adipocytes
Finny Monickaraj, Sankaramoorthy Aravind, Pichamoorthy
Nandhini,
Paramasivam Prabu, Chandrakumar Sathishkumar, Viswanathan
Mohan and Muthuswamy Balasubramanyam
Natalia Perilla Hernández
Salomé Rave Diosa
Third semester of medicine
Molecular Biology

2. INTRODUCTION
Recent studies have shown the importance of fat tissue in telomere
shortening and its relationship with obessity and type 2 diabetes.
This study was made inducing oxidative stress in 3T3-L1 adipocytes to
test shortened telomeres, senescence and functional impairment. This
adipocytes showed ROS, DNA damage, mRNA and protein expression
of senescence and pro-inflamatory markers, telomere length and
glucose uptake.
The researchers wanted to study adipocytes rather than white blood
cells due to its reflection on target tissues.

3. ADIPOCYTES
Become from fibroblasts,
stores lipids such as
triglyceride and cholesteryl
ester, as an energetic
reserve.
There are two types:
•White: one fat vesicle. Stores
lipids as a long term
energetic reserve.
•Brown: multiple vesicles.
Produces heat.

4. OXIDATIVE STRESS
Appears when there are
unpaired reactive oxygen
species (ROS) that generates
free radicals and hydrogen
peroxide which damages the
cellular components
including proteins, lipids and
DNA.
This damage lowers the ATP
levels leading to an
uncontrolled apoptosys that
releases cytotoxic
components.

5. INSULIN RESISTANCE
Its a genetic or acquired cell
inability to uptake insulin-
dependent glucose in tissues
such as liver, muscle and fat.
It leads to hyperglycemia
and disminished glucose
uptake if is not treated.
Some risk factors are obesity
and low physical activity.

6. ADIPOCYTE

7. OBJECTIVE
•Determine how oxidative stress and accelerated
senescence impacts fat tissue function and how
this, in turn, leads to age-related diseases.

8. MATERIALES Y MÉTODOS
•DMEM
•IBMX, insulina, dexametasona y FBS
Cultivo
•H2O2 - medio sin suero
Tratamiento de células e inducción de •H2O2 - glucosa oxidasa
estrés oxidativo •ADMA
•HG-LG
Tinción β galactosidasa asociada a •PBS
senectud
(SA – β-gal)
•Tinción DCF-DA
•HEPES
Medición ROS •PMA ROS
•PBS – agarosa – extendido – lisis – electroforesis – neutralización – tinción –
microscopio.
Ensayo cometa

9. MATERIALES Y MÉTODOS
Medición telómeros RT • DNA genómico – amplificación PCR –
- PCR relación T/S.
Cuantificación mRNA • cDNA – amplificación PCR – β actina.
RT-PCR
Extracción de proteínas • PBS – RIPA – Nano drop – SDS PAGE –
y Western blot Western blot – Ac – β actina.
• KRH – insulina – KRH – DOG – PBS –
Captación 2 – DOG SDS – conteo.

10. MATERIALES Y MÉTODOS

11. MATERIALES Y MÉTODOS
ABI 7000

12. MATERIALES Y MÉTODOS
W
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13. RESULTADOS

14. RESULTADOS

15. RESULTADOS

16. DISCUSSION
Morley. “In fact, diabetes mellitus has recently been Monickaraj agrees
2008 considered as a cause of accelerated aging”.
Tchkonia et al. “Despite the fact that senescence in adipocytes Monickaraj agrees
2010 could have profound clinical consequences because
of the large size of the fat organ and its central
metabolic role, there are only very few studies that
have looked at the senescence mechanisms in
adipocytes”.
Beliveau and Yaswen. “In fact, ectopic expression of hTERT was shown to Monickaraj agrees
2007 reduce the p53-dependent cellular stress
responses”.
Campisi. “Senescent cells are known to secrete molecules Monickaraj agrees
2005 that can alter the local microenvironment, such as
pro – inflammatory cytokines”.
Minamino and
Komuro.
2007

17. CONCLUSIONS
•Adipocytes subjected to glucose levels oscillation causes a
hyperactivation of p53 which is associated with insulin resistance, pro-
inflamatory effects and premature cell-aging.
•Adipocytes exposed to oxidative stress showed increased cellular
aging due to the following:
•Telomere shortening
•Increased expression of mRNA of p53 and p21
•Decreased adiponectin expression.

18. CONCLUSIONS
•The oxidative stress cause a increased secretion of IL6,
TNFa and decreased secretion of adiponectin and that
woul be reflected in insulin resistance.
•Type 2 diabetes and obesity induce oxidative stress
which causes a shortened telomeres, aging and
functional impairment

19. CONCEPTUAL MAP
Induced: NATALIA
H202, HG-LG, Obesity and
NATALIA
ADMA,GO type 2 diabetes
Increase
Oxidative p21
Insuline Aging
stress
resistance
Increase • Shortened
Dcreased ROS telomeres
p53
adiponectin • Functional
impairment
Pro-
inflamatory
cytokines Damage
Hyperglycemia
of:
DNA
PROTEIN Apoptosis
Increase LIPIDS
TNFa, IL6

20. CONCEPTUAL MAP
SALOME

21. GRACIAS
THANKS