SEMINARIO BIOMOL SUSANA SILVA Y STEPHANIE SERNA

Seminario Biología celular
Susana Silva
Stephanie Serna

1. Stephanie Serna Arbelaez – Susana Silva Salazar
Medical students – 3rd semester
Universidad Pontificia Bolivariana.

2. Melatonin is a hormone produced in the pineal gland.
Regulates the circadian rhythm.
Necroptosis is a form of celular death, different from
apoptosis. Mediated by RIPK1, RIPK3 y MLKL.
mtDNA is the DNA finded in the mitochondria.
INTRODUCTION

3. MELATONIN PROTECTS
LUNGS BY INHIBITING
MTDNA–STING–NECROPTOSIS
PATHWAY Sepsis → mtDNA release →
STING activation.
STING → necroptosis →
inflammation and lung
injury.
Melatonin blocks this
pathwayC → lung
protection.

4. GENERAL OBJECTIVE To examine whether melatonin alleviates sepsis-
induced acute lung injury by inhibiting
necroptosis through reducing the release of
circulating mtDNA.

5. ELISA
Inflammatory cytokines
measured in serum and
BALF

6. WESTERN
BLOT Necroptosis and STING pathway
proteins analyzed

7. INMUNOHISTOCHEMISTRY
Histology and inmunologic methods to detect especific
proteins
Detection of p-MLKL (final executor in the necroptosis) => IHC staining made possible
to visualize if lung cells of septic mice were activating necoptosis => Compare in
experimental groups.
IHC allowed to view how and where the
necroptosis occurred in the lungs, and
how the melatonin exerts its protective
effect.

8. qPCR
Measures in real time the amount of amplified
DNA in each cycle by using fluorescent signals.
It was used to identify the regions that
activate the STING cascade.
qPCR primers in different places of mtDNA and a nuclear control:
D-loop, mt-Cox 1, mMito, 18S rDNA.
Cuantification of mtDNA levels

9. SEPSIS INDUCES LUNG
INJURY AND NECROPTOSISFigure 1. Panels A & J

10. MELATONIN INHIBITS
NECROPTOSIS AND
PROTECTS LUNG TISSUE Figure 3. Panels A & E

11. RESULTS Figure 4. Panel E

12. RESULTS Figure 5. Panel A & E

13. DISCUSSION

14. CONCLUSIONS Identify the hidden
processes of
diseases Helps to see the efects even
unexpected compounds, like in the
case of melatonin that is primarly for
sleep and avoid necroptosis. Alternatives of treatment
through understanding
molecular processes

15. MAPA CONCEPTUAL SUSANA https://cards.algoreducation.com/app/set/68b46b8bd52144eac8686cca?
sheetId=68b46b8bd52144eac8686ccd&toolType=map

16. MAPA CONCEPTUAL
STEPHANIE