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Effects on lipid levels and lipid metabolism under more human-like conditions.

Aim of the study: Effects of garlic on lipid metabolism and atherosclerosis. Effects on lipid levels and lipid metabolism under more human-like conditions. Lipid levels, cholesterol synthesis and cholesterol absorption.

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Effects on lipid levels and lipid metabolism under more human-like conditions.

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  1. Aim of the study: Effects of garlic on lipid metabolism and atherosclerosis • Effects on lipid levels and lipid metabolism under more human-like conditions. • Lipid levels, cholesterol synthesis and cholesterol absorption. • Effects on fast atherosclerosis development and other processes that might be related with atherosclerosis. • Inflammation, blood pressure, platelet activation and atherosclerosis analysis.

  2. APOE*3-Leiden transgenic mouse as a model for hyperlipidemia and atherosclerosis studies • Transgenic mice carrying a mutant form of human APOE*3 gene. • Dominant dysfunctional APOE*3-Leiden results in defect in chylomicron/ VLDL clearance and more human-like lipoprotein profile. • Highly responsive to feeding cholesterol-rich diets. • Develops atherosclerosis varying from fatty streaks to more human-like complicated lesions.

  3. Effect of garlic on lipid metabolism and atherosclerosis in APOE*3-Leiden mice: Selection of compounds • Lichtwer LI111 garlic powder: Reference dry garlic powder. • Morado de Cuenca 200 (Mor200): Garlic powder rich in organic sulfur-containing compounds. • Diallyl disulfide (DADS): Isolated garlic compound rich in sulfur. • Allicin: Reference isolated garlic compound reported to result in 43% reduction of fatty streaks in C57Black/6 mice (D. Abramovitz et al., Coronary Artery Disease, 1999, 10: 515-519)

  4. 80 Effect of garlic on lipid metabolism and atherosclerosis in APOE*3-Leiden mice: Experimental scheme 25 -2 0 Regular diet Western diet Atherogenic diet 6 14 Run in 8 10 12 16 18 20 22 2 4 Matching and start of the study 80 65 Weight and food intake 80 80 80 65 60 65 65 65 Blood sampling/ plasma isolation 80 80 65 65 65 65 65 60 80 Plasma lipid levels 80 65 80 65 65 65 65 Cholesterol lipoprotein profile p p p p p p Chol. synthesis/ Intestinal absorption p p p p p Marker for liver function p p p Liver lipid composition 15 60 Blood pressure 80 65 65 Platelet activation measurements 80 65 Inflammation markers 60 Atherosclerotic aorta lesions typing 60 5 Atherosclerotic aorta lesions area 60 5 Compound biostability p p p p p Inflammation and atherosclerosis related parameters General parameters Lipid metabolism related parameters

  5. Plasma cholesterol levels Regular diet Western diet Atherogenic diet mM Control 3.1 9.3 63 LI111 2.9 9.3 57 * Mor 200 3.0 9.1 44 DADS 2.9 8.3 70 * Allicin 3.5 9.2 73 Effect of garlic on plasma cholesterol levels under 3 dietary conditions *P< 0.05

  6. squalene 1.5 lathosterol lanosterol 1.0 0.5 Concentration (µmol sterol / mmol cholesterol) 0.0 Control LI111 Mor 200 DADS Allicin 40 campesterol b-sitosterol 30 20 10 0 Control LI111 Mor 200 DADS Allicin Effect of garlic on markers of cholesterol synthesis and intestinal absorption under 3 dietary conditions Regular diet Markers of cholesterol synthesis Markers of intestinal absorption *P< 0.05

  7. Effect of garlic on liver function under 3 dietary conditions Plasma alanine amino transferase Regular diet Western diet Atherogenic diet U/L Control 33.4 30.3 175 t LI111 104 33.2 34.4 t Mor 200 94.1 32.9 30.3 DADS 29.9 41.8 203 Allicin 25.2 28.2 171 tP=0.051

  8. 8 250 p=0.001 p<0.001 200 6 * 150 * SAA (µg/ ml) vWF (%PP) 4 100 2 50 0 0 LI111 Mor 200 DADS LI111 Mor 200 DADS Allicin Control Allicin Control Effect of garlic on Serum Amyloid A and von Willebrand factor concentrations under fast atherogenic conditions *P< 0.05

  9. Effect of garlic powders on atherosclerosis under more human-like conditions: Groups selection • Control • Primador FRANCE 2001: 0.64 g/kg b.w. of dry garlic powder with low sulphur content. • Primador FRANCE 2001: 6.4 g/kg b.w. of dry garlic powder with high sulphur content. • Kyolic (positive control): 200 mg/kg b.w. of powder rich in S-allylcysteine reported to result in 64% reduction of fatty streaks in New Zealand white rabbits. Campbell JH, Efendy JL, et al. J.Nutrition. 2001;131:1006S-1009S.

  10. Effect of garlic powders on atherosclerosis under more human-like conditions: Experimental design Diet T + 0.2% cholesterol 28 In weeks 0 Run in 26 4 8 12 16 20 24 Matching and start of the study 60 Weight and food intake 60 60 60 55 60 60 60 60 Blood sampling/ plasma isolation 60 60 60 60 60 60 60 55 Plasma lipid levels 60 60 60 60 60 60 60 p Cholesterol lipoprotein profile p p Marker for liver function p p p p p p p p SAA and Fibrinogen 55 60 60 vWf, t-PA and soluble ICAM 55 60 60 All blood stimulation assay 55 Aortic root lesions typing and area 5 55 Aortic arch area and lipids deposition 55 5 Aortic root lesions composition 55 5 Mice on standard chow diet 8 5 Liver lipid composition (p.m.) 55 Compound biostability p p Inflammation and atherosclerosis development related parameters General parameters Lipid metabolism related parameters

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