Simvastatine, Edaravone and Dexamethasone protect against kainate - induced endothelial cell damage Examine the direct effects of kainate on cultured cells of the BBB and to test three anti inflammatory and antioxidant drugs used in clinical practice, simvastatin, edaravone and dexamethasone, to protect against kainate-induced changes.

1. Presented by:
Elizabeth Velasquez Ramirez
Sarita Velez Oviedo
Third semester of medicine
Universidad Pontificia Bolivariana

2. Introduction

3. Endothelial damage
• Alterations in endothelium regulating
factors
• Proinflammatory, prothrombotic,
vasoconstriction, increased cell adhesion
and oxidative stress
• Central pathway in CNS diseases

4. Overall Objective
Examine the direct effects of kainate on cultured cells of the BBB
and to test three anti inflammatory and antioxidant drugs used in
clinical practice, simvastatin, edaravone and dexamethasone, to
protect against kainate-induced changes.

5. Methods: cells cultures
• Create an environment where cells can live and
grow, simulating their physiological, metabolic and
biochemical conditions.
• Rat brain endothelial cells, glial cells, pericytes
and co-culture BBB.
• Cell viability, excitotoxicity, BBB permeability.

6. Treatments
• Kainate → was used to induce hyper excitability.
• Triton X - 100 → reference compound to cause cell death.
• Hydrogen peroxide → measure ROS and sodium nitroprusside.
• Simvastatine, Edaravone and Dexamethasone → potential protective agents.

7. PCR
• It is a technique where a gene or DNA fragment is
amplified directly or an RNA fragment indirectly.
• RNA amplification.

8. Immunohistochemistry
• It is a technique used to
differentiate tissues where a
specific cell location or cell is
pre-colored for easy differentiation.
• Effect of kainate (K) on rat brain
endothelial cells and kainate in
conjunction with statins.

9. Results

10. Results

11. Results
RT-PCR

12. Results
Immunohistochemistry

13. Discussion
Authors Position
Basuroy S, Lefer CW,
Parfenova H.
“Although oxidative stress and the production of excess ROS contribute
to the damage in excitotoxicity, and kainate was described to increase
ROS production in porcine brain endothelial cells, in the present
experiments kainate had no effect on ROS production in our model”.
Tóth AE, Walter FR, Bocsik
A, Sántha P, Veszelka S,
Nagy L.
“Edaravone was also protective against kainate induced damage and it
reduced ROS levels and kainate-induced NO production in our
experiments. These data confirm our previous results on the protective
and antioxidant effects of edaravone on cultured brain endothelial
cells”.
Minami T, Okazaki J,
Kawabata A, Kawaki H,
Okazaki Y, Tohno Y.
“In addition to its barrier protecting effect dexamethasone decreased
NO production and the iNOS mRNA expression elevated by kainate in
brain endothelial cells”.

14. Conclutions
1. immunohistochemistry allows us to observe that kainate makes
proteins discontinuous at the cell border, which increases cell
damage.
2.The presence of kainate receptors was demonstrated on brain
endothelial cells thanks to the PCR test.
3.the cells cultures allow us create an environment where cells can
live and grow, simulating their physiological, metabolic and
biochemical conditions.