1. Rolf Gebhardt University of Leipzig Medical Fakulty Institute of Biochemistry EU-Project: Garlic & Health Effects of High Quality Garlic on Markers of Arteriosclerosis and Inflammation in vitro and in vivo

2. Garlic & Health:Health part - general aim Elucidation of mechanism Analysis of active of action compoundsImproved drug formulation High quality garlic Improvement of garlic Optimal use of garlic plants and growth conditionsfor therapeutic application Main Focus:- Inflammation and cytokine signaling - Atherosclerosis and serum cholesterol - Cancer and biotransformation - Human Intervention Study Main Goal:- Elucidation of garlic action - Interactive improvement of garlic quality

4. Effects of Garlic on Cytokine Production Induced by LPS in Blood Cells: H.-P. Keiss, V. Dirsch, A. Vollmar University of Munich

6. Cholesterol Biosynthesis is highly regulated at various steps among others HMGCoA reductase is a key enzyme Many important biomolecules are derived from intermediates of cholesterol biosynthesis -->the biosynthetic pathway should never be completely inhibited

7. Comparison of different garlic-derived organosulfur compounds for inhibition of cholesterol biosynthesis

8. Inhibition of HMGCoA reductase and cholesterol biosynthesis Test Compound EC 50 values HMGCoACholesterol reductasebiosynthesis Allicin> 300 17 Diallyl disulfide> 250 65 Allyl mercaptan> 1500 450 S-allyl cysteine n.d. > 1500 Diallyl sulfide> 1000 735 Dipropyl disulfide> 1500 > 1000

9. Schematic view of the probable mode of action of several garlic-derived organosulfur compounds

10. Correlation between AMPK activity and inhibition of cholesterol biosynthesis Correlation coefficient: r = - 0.96 for all conditions: - absence of DADS (closed symbols) - presence of DADS (open symbols)

11. Central Role of AMP-dependent kinase (AMPK) as a mediator of stress signals, e.g. low energy charge Energy consuming pathways e.g. fatty acid and cholesterol synthesis are shut down

12. AMPK mediates the effect of DADS on various metabolic functions DADS + Inhibition of cholesterol and lipid biosynthesis + AMPK Cellular Stress ATP / AMP + antidiabetic effects

13. Studies on Lipid Metabolism in vivo: Use of transgenic ApoE Leiden mice as a model system for arteriosclerosis S. Espirito-Santo, H. Princen TNO Leiden, The Netherlands

16. Studies on Arteriosclerosis: K. Meyer, R. Gebhardt, S. Espirito-Santo, H. Princen

17. MMP TIMP ArteriosclerosisFibrosis ArthritisDiabetic Nephro- Rheumatoid Arthritis pathy InflammationGlomerulosclerosis Tumour-GrowthScleroderma -Metastasis -Angiogenesis Ulcers Parodontosis Morphogenesis, Differentiation, Wound healing, etc. Pathological Implications of MMP-TIMP-Imbalance:

18. Inhibition of MMP-9 and TIMP-1 Production by DADS, AM and SAC MMP-9 (activity assay) TIMP-1 (ELISA) HUVEC stimulated with PMA, forskolin and TNF-alpha

20. Summary: 1. Garlic substances like DADS show antiinflammatory modulation of cytokine production 2. DADS may inhibit hepatic cholesterol biosynthesis in certain diseased states 3. Garlic did not alter lipid profiles in vivo 4. DADS did modulate MMP-production by endothelial cells but again: in vivo no antiarteriosclerotic effects could be demonstrated