Workshops W2 Genes and environment: nutrigenetics W02-P-006 / HERBAL TERPENOIDS ACT AS LIGANDS FOR PPAR-ALPHA AND GAMMA TO MANAGE GENE EXPRESSION INVOLVED IN LIPID METABOLISM AND INFLAMMATION T. Kawada 1, N. Takahashi 1 , T. Goto 1, K. Egawa 2, S. Kato 1, K. Kuroyanagi 1, T. Kusudo 1, C. Kim 2, R. Yu 2. 1Division of Food Science and Biotechnology, Kyoto University, Uji, Japan; 2Department of Food Science and Nutrition, University of Ulsan, Ulsan, South Korea Objective: Terpenoids, which are contained in many herbal plants, show many biological effects: for example, anti-diabetic and anti- hypercholesteremic effects. Therefore, terpenoids have been thought valuable compounds to control medical conditions in vivo. However, a raechanism of the biological effects remains unclear. To address this issue, we examined the PPARalpha and gamma activities of terpenoids in cellurat function level and reporter assay. Methods: Anti-inflammatory effects were evaluated by the suppressed expression of TNF-alpha and COX2 in LPS-stimulated RAW264.7 macro- phages. Adipogenic effects were evaluated in 3T3-L 1 adipocytes. The effect on hepatic lipid metabolism were estimated by gene expression in HepG2 cells. PPARs lucfferase reporter assays were performed using an advanced highly sensitive system with coexpression of coactivator cAMP-response dement binding protein (CREB)-binding protein (CBP) developed by mod- ifying the dual luciferase system. The profiles of PPARalpha and gamma target genes by ligands were analyzed by a real time PCR in mcrophage, adipocyte and hepatocyte. Results: Abietic acid, one of terpenoids, strongly suppressed expression of TNF-alpha and COX2 in macrophages. This effect was resembled to that of a PPARgamma-synthetic ligand. The activity of abietic acid caused induction of PPARgamma-target gene expression in macrophages and adipocytes. Phytol, a component of chlorophyl, significantly enhanced the mRNA expression of acyl-CoA synthetase, carnitine palmitoyl trans- feraselA and acyl-CoA oxidase in HepG2. Actually, abietic acid and other terpenoids activated PPARalpha and/or gamma in reporter assays. Conclusions: Some terpenoids, such as abietic acid and phytol, act as naturally accruing PPARalpha and/or gamma ligands for not only anti-inflammation but also managing lipid metabolism and atherosclerosis. W02-P-007 ] -204C ALLELE OF THE CHOLESTEROL-7ALPHA HYDROXYLASE (CYP7A1) GENE DETERMINES HYPERRESPONSIVENESS OF LDL-CHOLESTEROL CONCENTRATION TO A HIGH-FAT DIET J. Kov~r 1, D. Bobkov~ 1, P. Such~nek 1, J. A. Hub~icek 1, M. Rudling 2, R. Poledne 1. l lnstitute for Clinical and Experimental Medicine, Prague, Czech republic; 2Karolinska Institute at Huddinge University Hospital, Stockholm, Sweden Objective: The data from population studies brought the evidence that A-204C polymorphism in the cholesterol-7a hydroxylase (CYP7A 1) gene is associated with the cholesterolemia responsiveness to a content of fat and/or cholesterol in a diet. Therefore, to test the hypothesis that -204C promotor variant of CYP7A 1 gene is responsible for hyperresponsiveness of choles- terol and LDL-cholesterol concentration to a diet rich in fat and cholesterol was tested in a dietary intervention study in young healthy volunteers. Methods: Eleven men homozygous for -204A allele (AA subjects) and thirteen homozygous for -204C allele (CC subjects) were included into the study. They did not differ in age, body mass index, lipid concentration and apo E gene polymorphism at the entry into the study. They were fed by the high-fat high-cholesterol (HF) diet for three weeks and by the low-fat low-cholesterol (LF) diet for another three weeks in randomized order in cross-over design. LF diet contained 22% of fat (one third of that being animal fat) and 2.2 mg of cholesterol/kg of body weight, HF diet contained 40% of fat (86% of animal fat) and 9.7 mg of cholesterol/kg. At the end of each dietary period, blood was taken after 12hour fasting and lipeproteins were isolated by ultracentfifugation; the concentration of 7ct-hydroxy-4-cholesten-3-one (C4) was also determined. Remits: In CC subjects, cholesterol, LDL-cholesterol, and apo B concentrations were significantly higher on HF diet than on LF diet (12%, 22%, and 8%, respectively), whereas no such difference was observed in AA subjects. That suggests that CC subjects are hyperresponsive to HF diet. Interestingly, no difference in change of C4/cholesterol ratio (that correlates with CYP7A1 activity) between both groups between LF and HF diet was observed. Conclusions: Our data.bring an evidence that -204C allele but not -204A allele is involved in detemaining the hyperresponsiveness of LDL- cholesterol to a diet rich in fat and cholesterol. However, the exact mechanism involved remains to be determined. The study was supported by grant # 1M6798582302 from ME CR. wo2-P-OOSJ SYNERGISTIC EFFECT OF STROMELYSIN-1 (MATRIX METALLOPROTEINASE-3) PROMOTER 5A/6A POLYMORPHISM WITH SMOKING ON THE ONSET OF ACUTE CORONARY SYNDROME P.-Y. Liu 1, J.-H. Chen 1 , Y.-H. Li 1, H.-L. Wu 2, G.-Y. Shi 2 . 1Division of Cardiology, Internal Medicine, National Cheng-Kung University Hospital, Tainan, Taiwan; 2 lnsitute of Biochemistry, College of Medicine, National Cheng-Kung University, Tainan, Taiwan Background: Plaque rupture with thrombosis is well established as a critical factor in the pathogenesis of acute coronary syndrome (ACS). Stromelysin-1, also called MMP-3, can degrade extracellular matrix and may contribute to the weakening of the cap and subsequent rupture. Methods: We studied 350 consecutive patients diagnosed as ACS (with 250 ~I, 100 NSTMI and 100 unstable angina patients, respectively) with mean age of 55.7-t-8.5 years (86% men) and 200 sex-matched control subjects (mean age 57.55=7.8 years) of 5A/6A mutation at stromelysin-1 promoter by using PCR and direct DNA sequencing. Results: The frequency of the 5A mutation (SA/5A + 5A/6A genotypes) was significantly higher among ACS than the control group (29% vs. 18%, OR 2.35, 95% CI 1.2 to 6.6, p=0.01). The baseline characters among 3 subgroups of ACS were similar except with lower rate of DM in STEMI group. Multiple logistic regression analysis showed that the 5A allele polymorphism was an independent risk factor (OR 2.15, 95% CI 1.2 to 5.3, p=0.01) as were as smoking (OR 3.77, 95% CI 1.5 to 5A, p=0.001), DM (OR 3.51, 95% CI 1.4 to 6.3, p=0.003) and hypertension (OR 1.87, 95% CI 1.9 to 7.8, p=0.001) for the onset of ACS. Moreover, among patients who did not smoke, the 5A allele polymorphism was associated with an increase in the risk of ACS (OR 3.95, 95% CI 1.3 to 9.3). Furthermore, smoking carriers of the stromelysin-1 5A allele polymorphism had a significantly 9-fold increased risk of ACS (OR 8.75, 95% CI2.5 to 15.5) when compared with non-smoking non-carriers. Conclusion: There was a significant association between the 5A/6A polymorphism in the promoter region of stromelysin-1 gene and ACS in Taiwan. Both the 5A/6A polymorphism of stromelysin-1 gene and smoking are independent risk factors for ACS population. A synergistic effect between these two risk factors for ACS had been shown in this study. WO2-P-O09] THE APOLIPOPROTEIN E GENE PROMOTER -219G/T POLYMORPHISM DETERMINES INSULIN SENSITIVITY IN RESPONSE TO DIETARY FAT IN HEALTHY YOUNG PEOPLE J.A. Moreno, J. L6pez-Miranda, P. P6rez-Maxtinez, R. Moreno, P. G6mez, A. Lozano, B. Cort6s, R.A. Fem~indez de la Puebla, J. Delgado, F. Fuentes, E P6rez-Jim4nez. Lipids and Atherosclerosis Research Unit. Hospital Universitario Reina Sofia, C6rdoba, Spain Objective: To determine whether the APOE gene promoter (-219Gfl') polymorphism is related with significantly different insulin sensitivity in response to changes in the quantity and quality of dietary fat in healthy subjects. Methods: 59 volunteers (10 GG, 36 GT and 13 "Iq') were subjected to three dietary periods, each lasting four weeks. The first was a saturated fatty acid (SFA)-rich diet (38% fat, 20% SFA), followed by a carbohydrate (CHO)-fich diet (30% fat, 55% carbohydrate) or a monounsaturated fatty acid (MUFA)-rich diet (38% fat, 22% MUFA) following a randomized crossover design. For each diet, we investigated peripheral IS with the insulin suppression test. Results: Steady-state plasma glucose (SSPG) concentration was lower (p<0.05) in GG subjects as compared with GT and "IT individuals, inde- pendently of the diet consumed. A significant diet-by-genotype interaction effect was observed on SSPG and non-esterified free fatty acid (NEFA) plasma levels. Thus, the shift from MUFA- or a CHO-rich diets to the SFA-rich diet increased (p<0.05) SSPG and NEFA concentration in GG and GT subjects, but not in "IT volunteers. Conclusion: Carriers of the -219T allele have lower IS than GG individ- uals. Furthermore, only carriers of the -219G allele have an improvement in 75th EAS Congress, 23-26 April 2005, Prague, Czech Republic 0 I 0 03