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Effects of Electromagnetic Pulses on the Permeability of Blood-Brain-Barrier

Effects of Electromagnetic Pulses on the Permeability of Blood-Brain-Barrier. Gui-Rong Ding, Ph.D Department of Radiaition Medicine, Fourth Military Medical University, China. Introduction.

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Effects of Electromagnetic Pulses on the Permeability of Blood-Brain-Barrier

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  1. Effects of Electromagnetic Pulses on the Permeability of Blood-Brain-Barrier Gui-Rong Ding, Ph.D Department of Radiaition Medicine, Fourth Military Medical University, China

  2. Introduction • EMP( Electromagnetic pulses) exposure resulted in various changes in morphology and function of nervous system. • BBB (Blood-Brain-Barrier) plays an important role in maintaining homeostasis in brain, and BBB disruption can initiate the development of brain injuries. • Our previous study indicate that membrane is one of the sensitive targets of EMP.

  3. Blood–Brain Barrier and the Tight Junction Endothelium Tight junction Basal Lamina Astrocytic Foot Process

  4. Our concern ? EMP exposure BBB openning Molecular mechanism?

  5. Material and Methods Animals SD rats, male,6-8 weeks old，200±20g

  6. Material and Methods Exposure apparatus EMP Generator GTEM-Cell

  7. Material and Methods EMP parameters • Frequency band 0-1.5 GHz • Half width 100ns • Pulses number 50 , 100 and 200 • Intensity 50 , 100, 200 and 400 kV/m

  8. Material and Methods BBB permeability detection Serial frozen section, whole coronal slices of brain Evans Blue (EB) tracer and fluorescent microscope Lanthanum nitrate tracer and transmission electron microscope Protein expression Western blotting Immunofluorescence

  9. control 50kV/m, 200 pulses 100kV/m, 200pulses Results EMP induced BBB opening (dose-effect) 200kV/m, 200pulses

  10. Results EMP induced BBB opening (dose-effect) 200kV/m, 50 pulses 200kV/m, 100pulses 200kV/m, 200pulses

  11. 2h after exposure 4h after exposure 6 h after exposure 200kV/m, 200pulses Results EMP induced BBB opening (time-effect)

  12. EMP induced BBB opening 200kV/m 200pulses (×10000) control (×10000) Results

  13. Changes of BBB tight junction related proteins after EMP exposure ZO-1 Occludin Claudin-5 3h after exposure(200kV/m, 200pulses) Control

  14. Protein expression of tight junction related proteins after EMP exposure 220kDa Cellular fractionation ZO-1 ZO-1 220kDa Claudin-5 22kDa Occludin 65kDa Occludin 65kDa SOL CSK SOL CSK SOL CSK SOL CSK CSK Con Con 0hr 3hr 6hr 6hr 0hr 3hr SOL Membrane protein

  15. Conclusions • EMP exposure may lead to an increase in the permeability of BBB in rats. • The injury of BBB tight junction was involved in EMP-induced BBB permeability increase.

  16. Ongoing work Primary cultured brain microvascular endothelial cells(BMEC) Primary cultured brain astrocyte

  17. In vitro BBB model Polyester-multiple-membrane Astrocytes BMEC Transwell Transwell Astrocytes Transwell Culture dish Culture dish

  18. Blood Testicle Barrier

  19. Effects of EMP (200kV/m, 200 pulses) on the Permeability of Blood-Testicle -Barrier control 1h 6h 2h

  20. Changes of BTB tight junction related protein-ZO-1 after EMP (200kV/m, 200 pulses)exposure 1h control 2h 6h

  21. Further research • To set up in vitro BBB model • To identify the size of the molecules that get though BBB after different parameters EMP exposure • To clarify the molecular mechanism of EMP-induced BBB opening

  22. Further research • By observing the permeability and changes of TJs in other “Barrier systems”, to find out the common mechanism of EMP-induced barrier injury. BBB In common Sensitive organs Barrier system BRB EMP exposure Tight Junctions（TJs） BTB

  23. Thank you!

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