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Prof. A. M. Vollmar PD Dr. V. M. Dirsch Dr. Hans Peter Keiss Thomas Roos Nancy Lopez

WP 5: Cardiovascular diseases 5.1: The Interaction of garlic with mediators of inflammation. Prof. A. M. Vollmar PD Dr. V. M. Dirsch Dr. Hans Peter Keiss Thomas Roos Nancy Lopez. Highlights. Specific Targets:. Garlic. Adhesion: CAM expression. Endothelial activation.

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Prof. A. M. Vollmar PD Dr. V. M. Dirsch Dr. Hans Peter Keiss Thomas Roos Nancy Lopez

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  1. WP 5: Cardiovascular diseases 5.1: The Interaction of garlic with mediators of inflammation • Prof. A. M. Vollmar • PD Dr. V. M. Dirsch • Dr. Hans Peter Keiss • Thomas Roos • Nancy Lopez Highlights

  2. Specific Targets: Garlic Adhesion: CAM expression Endothelial activation Migration: Cytokines Cytokines Smooth muscle cell growth Aktivation NF-kB Activated macrophage

  3. Objectives • Influence of „garlic“ on the activation of the • transcription factor NF-kappaB • Influence of „garlic“ on the expression of adhesion • molecules (ICAM-1, VCAM-1, E-Selectin) • Influence of „garlic“ on cytokine production in blood cells

  4. Transcription factor NF-kB activation P proinflammatory factors i.e. CAM, Cytokines TNF-a P IkB NF-kB binding activity transactivation activity

  5. + radio-labeled oligonucleotides isolation of nuclear protein gel-electrophoresis autoradiography EMSA: electrophoretic mobility shift assay

  6. NF-kappaB luciferase reporter-gen assay luciferase TNF-a lysis activation luciferase light + substrate luciferase reporter- gen Luminometer

  7. Measurement of adhesion molecules Analysis by FACS (fluorescence activated cell sorting) Fluorochrome-labeled antibodies against Test compound e.g. ICAM-1, VCAM-1, E-Selectin TNF-a ICAM-1 control TNF-a/ 8 h Endothelial cells (HUVECS)

  8. Eur J Nutr., in press

  9. cytokine Cytokines / Chemokines (blood cells) supernatant: blood medium test compound = garlic LPS 37°C, 5% CO2,16 h ELISA (Enzyme –Linked-ImmunoSorbent-Assay)

  10. J. Nutr., 2003, 133: 2171-2175

  11. Additional targets • Effect of garlic on NO production • Effect of garlic on PAI • Effect of garlic constituents on VSMC proliferation

  12. Garlic induced NO-liberation Garlic constituens 1.5 S-allylcysteine 1.0 Diallyl disulfide x-fold Allylmethyl-SO 2 Allicin 0.5 Allylmethyl-sulfide Allyl-mercaptan 0.0 1.25 CN 10 µM 100 µM 1.00 0.75 Spain 2000 x-fold France 2000 Spain 2001 0.50 France 2001 0.25 0.00 CN 50 µg/ml 500 µg/ml Printanor

  13. Garlic and PAI-1 and tPA

  14. + Serum ? VSMC proliferation garlic Rat Primary rat aortic vascular smooth muscle cells (VSMC)

  15. n.s. n.s. n.s. 750000 500000 Number of cells 250000 serum 10% - + + + + 25 50 100 DADS (µM) Cell proliferation - DADS

  16. CS-activated cells Quiescent cells 25 µM DADS + CS 50 µM DADS + CS Events 100 µM RV + CS 100 µM DADS + CS DNA content Cell cycle analysis

  17. Outlook for the „real“ highlight Human intervention study

  18. Special highlight Dr. rer. nat. Hans- Peter

  19. Special highlight PD Dr. Verena Dirsch Professorship in Vienna

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