HEMORRHAGIC SHOCK INDUCED FLUID LEAK AND MICROVASCULAR ENDOTHELIAL - - PowerPoint PPT Presentation

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HEMORRHAGIC SHOCK INDUCED FLUID LEAK AND MICROVASCULAR ENDOTHELIAL - - PowerPoint PPT Presentation

May 17, 2023 168 likes •392 views

DOXYCYCLINE INHIBITS HEMORRHAGIC SHOCK INDUCED FLUID LEAK AND MICROVASCULAR ENDOTHELIAL DERANGEMENTS Katie Wiggins-Dohlvik, Hayden Stagg, Himakarnika Alluir, Chinchusha Anasooya Shaji, Matthew Davis, Binu Tharakan Nothing to Disclose

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Katie Wiggins-Dohlvik, Hayden Stagg, Himakarnika Alluir, Chinchusha Anasooya Shaji, Matthew Davis, Binu Tharakan

DOXYCYCLINE INHIBITS HEMORRHAGIC SHOCK INDUCED FLUID LEAK AND MICROVASCULAR ENDOTHELIAL DERANGEMENTS

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SLIDE 2

Nothing to Disclose

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Hemorrhage

  • Treatment:
  • Stop the bleeding and fill the

tank

  • But is it that simple…
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Hemorrhage

Bench-to-bedside review: Latest results in hemorrhagic shock Angele MK, Schneider CP, Chaudry IH - Crit Care (2008)

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SLIDE 5

0 minutes 30 minutes 60 minutes

Sham HS

Vessel Permeability

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SLIDE 6
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SLIDE 7

On a microscopic level, the Adherens Junction…

www.humpath.com

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SLIDE 8

Background

  • Degrade ECM
  • Degrade AJ

proteins

Matrix metalloproteinases

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SLIDE 9

Background

  • Increase vascular

permeability

  • Mechanism not

fully understood

ROS (Reactive Oxygen Species)

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SLIDE 10

Background Doxycycline Anti MMP and ROS outside HS

  • AAA Aziz 2007
  • HTN/CV disease Castro 2013
  • Re-absorptive bone

diesase Franco 2011

  • Trichiasis Li 2013
  • Leukemia Wang 2015
  • Burn Stagg 2013
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Hypothesis

HS

Fluid Leak

ROS MMP

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Serum Collected for in vitro experiments

HS Sham

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Experiments

  • Rat Lung Microvascular Endothelial Cells
  • 4 Groups:
  • Sham
  • HS
  • Sham + doxycycline (.2ug/ml)
  • HS + doxycycline (.2ug/ml)
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SLIDE 14

Permeability

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SLIDE 15

FITC-Albumin Fluorescent Intensity (% of Sha m)

* Sham vs HS, p<0.05 § HS vs HS Doxycycline, p<0.05

Permeability

*

§

20 40 60 80 100 120 140 160 180 Sham Shock Doxycycline + Sham Doxycycline + Shock HS Sham + Doxycycline Sham HS + Doxycycline

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ROS Formation Assay

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100 200 300 400 500 600 700 800

Sham Shock Doxycycline +Sham Doxycycline +Shock

ROS formation (% of Sham)

Sham HS + Doxycycline HS Sham + Doxycycline

ROS Formation

* Sham vs HS, p<0.05 § HS vs HS Doxycycline, p<0.05 §

*

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SLIDE 18
  • Cells stained for:

– β-Catenin – f Actin

Adherens Junctions

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SLIDE 19

Staining for β-Catenin

Adherens Junctions

HS Sham

Sham + Doxycycline HS + Doxycycline

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Staining for f-Actin

Adherens Junctions

Sham + Doxycycline HS Sham HS + Doxycyclin e

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Discussion

  • HS
  • Increases vascular permeability
  • Increases permeability
  • Increases ROS formation
  • Damages adherens junctions
  • Doxycycline attenuates

changes in vitro

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SLIDE 22

What’s next?

  • Assay MMP-9 levels
  • Examine Doxycycline in vivo
  • And beyond…