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LISSETH KARINA MARÍN RÍOS
3RD SEMESTER
UNIVERSIDAD PONTIFICIA BOLIVARIANA
2024
A venous thrombosis is a serious condition that
occurs when a blood clot forms in a vein,
typically in the legs, thighs, or pelvis. These clots
can block blood flow and cause swelling, pain,
and even break off and travel to the lungs,
leading to a life-threatening condition called
pulmonary embolism.
VENOUS THROMBUS
INTRODUCTION
Function:
Acts as both a pro-inflammatory cytokine and an anti-
inflammatory myokine (produced by muscle cells)
Production:
Primarily produced by immune cells like macrophages and T cells,
but also by other cell types like endothelial cells and fibroblasts
Effects:
Stimulates the production of acute phase proteins in the liver
during inflammation
Plays a role in fever and the acute phase response
Regulates various metabolic processes
IL-6
INTRODUCTION
Analyzing experimental
resolution of venous
thrombus is driven by IL-6-
mediated monocyte
actions.
GENERAL
OBJECTIVE
The objective of this experimental method is to investigate how
the protein composition of blood clots (thrombi), the inferior
vena cava (IVC), and specific immune cells (BMDMs and
RAW264.7) is influenced by exposure to IL-6 (an inflammatory
molecule) or Brefeldin A (an inhibitor of protein transport).
The objective of this method is to analyze the
structure, cellular composition, and components of
blood clots (thrombus) formed in the lower back
(retroperitoneal area) of mice.
METHODS
WESTERN BLOTS
HISTOLOGY
The objective of this experimental method is to
measure and analyze the levels of specific RNA
molecules within cells
METHODS
FLOW CITOMETRY
PCR
The objective of this experimental method is to isolate and
analyze specific types of immune cells from mice to
investigate how they are involved in the process of blood
clot formation or resolution
Following sham surgery
or stasis thrombosis, vein
wall lysate protein levels
Gel blots cropped from
different parts of the gel are
presented, with white line
separation. Protein levels of
intrathrombus.
Representative
photomicrographs at
40 × of WT (left, arrows
mark recanalization
channels) and IL-6−/−
RESULTS
Quantification of protein
levels relative to total
protein loading is shown
in the upper panels.
Protein levels of MMP9 and
Urokinase were assayed in the
BMDMs
DISCUSSION
From this series of experiments, we
have characterized the role of IL-6 in
two models of VT which replicate
clinical venous thrombosis; namely,
partial and complete blood stasis.
We found that IL6 gene deletion did
not affect thrombus formation,
contrary to data suggesting that anti-
IL-6 inhibition can impair
thrombogenesis
. Zhang, Y. (2020)
. Deatrick,K.B. (2011)
IL-6 has been shown to play a role in
thrombogenesis and resolution17,54,
but not as a key mediator of fibrinolysis
. Stouthard,J.M. (1996)
AUTHOR
What did they say?
AGREE OR
DESAGRREE
Conclusions
Interleukin-6 (IL-6) plays a crucial role in the resolution of venous thrombi:
The article demonstrates that IL-6 is produced by macrophages at the thrombus site.
IL-6 promotes the differentiation of monocytes to macrophages with a pro-resolving
phenotype. These pro-resolving macrophages phagocytize (ingest) dead cells and debris,
which facilitates dissolution of the thrombus.
Implications for the development of new therapies:
Strategies that modulate IL-6 signaling or that increase the activity of pro-resolving
macrophages could be useful in promoting thrombus dissolution and preventing thrombotic
complications.
* TVP: Trombosis venosa profunda EP: Embolisis Pulmonar
Referencias: Mayo Clinic Trombosis venosa profunda (TVP) [Internet]. 2022. Disponible en: https://www.mayoclinic.org/es/diseases-conditions/deep-vein-thrombosis/symptoms-causes/syc-20352557
National Heart, Lung, and Blood Institute. Venous thromboembolism [Internet]. Disponible en: https://www.nhlbi.nih.gov/health/venous-thromboembolism

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Seminario Biología Molecular

  • 1. LISSETH KARINA MARÍN RÍOS 3RD SEMESTER UNIVERSIDAD PONTIFICIA BOLIVARIANA 2024
  • 2. A venous thrombosis is a serious condition that occurs when a blood clot forms in a vein, typically in the legs, thighs, or pelvis. These clots can block blood flow and cause swelling, pain, and even break off and travel to the lungs, leading to a life-threatening condition called pulmonary embolism. VENOUS THROMBUS INTRODUCTION
  • 3. Function: Acts as both a pro-inflammatory cytokine and an anti- inflammatory myokine (produced by muscle cells) Production: Primarily produced by immune cells like macrophages and T cells, but also by other cell types like endothelial cells and fibroblasts Effects: Stimulates the production of acute phase proteins in the liver during inflammation Plays a role in fever and the acute phase response Regulates various metabolic processes IL-6 INTRODUCTION
  • 4. Analyzing experimental resolution of venous thrombus is driven by IL-6- mediated monocyte actions. GENERAL OBJECTIVE
  • 5. The objective of this experimental method is to investigate how the protein composition of blood clots (thrombi), the inferior vena cava (IVC), and specific immune cells (BMDMs and RAW264.7) is influenced by exposure to IL-6 (an inflammatory molecule) or Brefeldin A (an inhibitor of protein transport). The objective of this method is to analyze the structure, cellular composition, and components of blood clots (thrombus) formed in the lower back (retroperitoneal area) of mice. METHODS WESTERN BLOTS HISTOLOGY
  • 6. The objective of this experimental method is to measure and analyze the levels of specific RNA molecules within cells METHODS FLOW CITOMETRY PCR The objective of this experimental method is to isolate and analyze specific types of immune cells from mice to investigate how they are involved in the process of blood clot formation or resolution
  • 7. Following sham surgery or stasis thrombosis, vein wall lysate protein levels Gel blots cropped from different parts of the gel are presented, with white line separation. Protein levels of intrathrombus. Representative photomicrographs at 40 × of WT (left, arrows mark recanalization channels) and IL-6−/− RESULTS
  • 8. Quantification of protein levels relative to total protein loading is shown in the upper panels. Protein levels of MMP9 and Urokinase were assayed in the BMDMs
  • 9. DISCUSSION From this series of experiments, we have characterized the role of IL-6 in two models of VT which replicate clinical venous thrombosis; namely, partial and complete blood stasis. We found that IL6 gene deletion did not affect thrombus formation, contrary to data suggesting that anti- IL-6 inhibition can impair thrombogenesis . Zhang, Y. (2020) . Deatrick,K.B. (2011) IL-6 has been shown to play a role in thrombogenesis and resolution17,54, but not as a key mediator of fibrinolysis . Stouthard,J.M. (1996) AUTHOR What did they say? AGREE OR DESAGRREE
  • 10. Conclusions Interleukin-6 (IL-6) plays a crucial role in the resolution of venous thrombi: The article demonstrates that IL-6 is produced by macrophages at the thrombus site. IL-6 promotes the differentiation of monocytes to macrophages with a pro-resolving phenotype. These pro-resolving macrophages phagocytize (ingest) dead cells and debris, which facilitates dissolution of the thrombus. Implications for the development of new therapies: Strategies that modulate IL-6 signaling or that increase the activity of pro-resolving macrophages could be useful in promoting thrombus dissolution and preventing thrombotic complications.
  • 11. * TVP: Trombosis venosa profunda EP: Embolisis Pulmonar Referencias: Mayo Clinic Trombosis venosa profunda (TVP) [Internet]. 2022. Disponible en: https://www.mayoclinic.org/es/diseases-conditions/deep-vein-thrombosis/symptoms-causes/syc-20352557 National Heart, Lung, and Blood Institute. Venous thromboembolism [Internet]. Disponible en: https://www.nhlbi.nih.gov/health/venous-thromboembolism