- Email: [email protected]
T polymorphism determines insulin sensitivity in response to dietary fat in healthy young people
Workshops W2 Genes and environment: nutrigenetics W02-P-006
/ HERBAL TERPENOIDS ACT AS LIGANDS F O R PPAR-ALPHA AND G A M M A TO MANAGE GENE EXPRESSION INVOLVED IN LIPID METABOLISM AND I N F L A M M A T I O N
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 antihypercholesteremic 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 macrophages. 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 modifying 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 transferaselA 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 A L L E L E OF T H E C H O L E S T E R O L - 7 A L P H A HYDROXYLASE (CYP7A1) GENE DETERMINES HYPERRESPONSIVENESS OF L D L - C H O L E S T E R O L C O N C E N T R A T I O N 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 cholesterol 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 LDLcholesterol 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-OOSJSYNERGISTIC EFFECT OF STROMELYSIN-1 (MATRIX METALLOPROTEINASE-3) P R O M O T E R 5A/6A P O L Y M O R P H I S M W I T H S M O K I N G 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; 2lnsitute 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 A P O L I P O P R O T E I N E GENE P R O M O T E R -219G/T P O L Y M O R P H I S M DETERMINES INSULIN SENSITIVITY IN RESPONSE TO DIETARY FAT IN HEALTHY Y O U N G P E O P L E 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, independently 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 individuals. 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
Workshops W2 Genes and environment: nutrigenetics
10
IS when a MUFA- or a CHO-rich diets are consumed instead of a SFA-rich diet.
W02-P-010 I FOLATE LEVELS D E T E R M I N E E F F E C T OF ANTIOXIDANT SUPPLEMENTATION ON M I C R O N U C L E I IN SUBJECTS W I T H CARDIOVASCULAR RISK B. Smolkov~ ~, M. Du~insk~i 1, K. Rasqovfi I , M. Barancokov~i I , A. Kafimirov~ 1, A. Horskh 1, V. Spnstovh 1, J. Gagparovic 1, A. Collins 2.
1Research Base of Slovak Medical University, Institute of Preventive and Clinical Medicine, Bratislava, Slovakia; 2Department of Nutrition, University of Oslo, Norway
Objective: We studied the effect of modest supplementation with c~tocopherol (100 mg/day), ~-carotene (6 mg/day), vitamin C (100 mg/day) and selenium (50 ~tg/day) on oxidative stress and chromosomal damage markers. Subjects and methods: Two groups of middle aged men differing in cardiovascular risk [28 survivors of myocardial infarction before age 50 (MI), and 58 healthy controls (RC)] were randomly divided into equal groups to receive antioxidants or placebo for 12 weeks. Micronucleus (MN) levels in peripheral lymphocytes, and changes seen after intervention, were studied in relation to MTHFR C677Tgenotype, basal plasma homocysteine (hey) and relate levels. Ferric reducing ability of plasma (FRAP) and concentration of malondialdehyde (MDA) were measured to assess the antioxidant effect of supplementation. Results: MN frequency was not significantly associated with folate, hey or MDA levels before supplementation. MN frequencies and plasma folate levels did not vary significantly with MTHFR genotype. Hcy levels in subjects w i t h / T variant genotype were significantly higher compared with CT or CC (17.094-5.96 vs. 11.934- 4.34 and 12.224- 3.84 p.mol/L; P=0.001), especially in subjects with low folate (21.77±8.16 vs. 9.84± 2.41 and 14.584- 3.93 mol/L; P=0.012). In the placebo RC group (but not in the supplemented group) an increase of MN (from 5.064-3.59 to 7.284-5.47cells with MN; P=0.00) was detected at the end of the intervention period. In antioxidant-supplemented MI survivors we found an increase in FRAP (from 1000±163 to 1306±167 p,mol/L; P<0.001) and a decrease in MDA (from 2.344-1.0 to 1.334-0.4 Ixmol/L; P=0.001). MN frequency showed a decrease, which was strongest in subjects with normal folate levels (from 8.70 4- 4.71 to 4.054-2.29 cells with MN; P=0.015). In subjects with low folate, a high correlation was found between MN after supplementation and hey both before (r=- 0.979, P=0.002) and after supplementation (r=-0.922, P=0.009). Conclusions: Folate deficiency may amplify the effect of other risk factors such as elevated lacy levels or variant MTHFR genotype, as well as influencing the ability of antioxidant supplementation to protect against genetic damage. I W02-P-011 I M A R K E R S OF OXIDATIVE DAMAGE TO LIPIDS AND DNA RELATED TO SEASONAL VARIATIONS IN DIET I
B. Smolkov~ 1, M. Du~insk~ 1, K. Ra~lov~iI , G. McNeill 3, V. Spustov~ 1, P. Bla~cek 2 , A. Horsk~i I , J. Gagparovic 1, A. Conins 4. 1Institute of
Preventive and Clinical Medicine, Slovak Medical University, Bratislava, Slovakia; 2Hospital of Ministry of Defence, Bratislava, Slovakla; University of Aberdeen, Scotland, UK; 4Department of Nutrition, University of Oslo, Norway
Objective: The relatively high incidence of cardiovascular disease and cancer in countries of centraYeastem Europe might be associated with nutritional imbalance, particularly in winter months when traditionally the supply of fresh fruit and vegetables is limited. Subjects and methods: We monitored markers of oxidative stress and antioxidants in three Slovak population groups: 46 survivors of myocardial infarction (MI), 48 healthy, normolipidemic subjects (NL) and 70 rural controls (RC). Data were collected in February/March (winter/spring-Wl, W2) and September/October (summer/autumn-A1, A2) of two consecutive years, representing times of minimum and maximum local availability of fresh fruits and vegetables. Lipid peroxidation was assessed by the concentration of malondialdehyde (MDA) in plasma and lymphocyte DNA oxidation was measured using the comet assay. Dietary antioxidants, folate levels, homocysteine and total antioxidant status were measured in plasma.
Results: Food frequency questionnaires showed that vegetable consumption in summer/autumn was twice as high as in winter/spring (P<0.001). DNA damage did not vary consistently across the seasons, but mean plasma MDA levels for the MI and NL groups showed a clear pattern, with high levels in winter/spring and low levels in summer/autumn (/VII group: 3.604-1.98, 1.394-1.00, 1.894-1.03, 0.904-0.38 pmol/L; NL group: 1.85+0.90, 0.90+0.20, 1.27+0.66, 0.90±0.24 p,mol/L for W l , A1, W2, A2; P<10-7). Folate levels varied in the opposite way and followed the pattern of vegetable consumption (MI group: 4.20±1.3, 5.60±1.3, 3.964-1.4, 4.764-1.4 ng/ml; NL group: 4.624-1.5, 5.924-1.5, 5.054-2.7, 6.17±2.2 ng/ml for W l , A1, W2, A2; P<10-s). The RC group had the smallest seasonal variations - in vegetable consumption, folate levels, and MDA. Conclusions: High winter MDA levels are seen in those individuals with relatively low folate; they never occur in subjects with high plasma folate, implying that folate levels might directly protect against lipid oxidation. This study illustrates the value of the molecular epidemiological approach, while emphasising the need for well characterised population groups and valid biomarkers.
I WO2-P-012 I i
THE A P O E 2 K N O C K - I N M O U S E AS A M O D E L F O R TYPE III HYPERLIPIDEMIA AND STEATOHEPATITIS
R.S. Sverdlov-Shiri 1, K. Wouters 1, P.J.J. van Gorp 1, M.J.J. Gijbels 1, B. Staels 3, M. van Bilsen 2, M.H. Hofker 1. 1Dept. of Molecular Genetics,
University Maastrich~ Maastricht, The Netherlands; 2Dept. of Physiology, University Maastrich~ Maastricht, The Netherlands; 3Institute Paster,Lille, France The apolipoprotein E2 (APOE2) allele is the main cause of Type III hyperlipidemia in man. The APOE2 knock-in (APOE2ki) mouse develops severe diet-induced hyperlipidemia and is highly responsive to fibrate, a PPAR~ agonist. Objective: Obtaining a better understanding of the molecular and physiological mechanisms underlying hyperlipidemia Methods: APOE2ki mice were fed a high-fat diet or a high-fat diet in combination with fenofibrate. The mice were sacrificed at several time points after start of the diet. As a control, we used chow-fed APOE2ki mice. In addition to physiological measurements, microarray analysis was preformed using liver-derived mRNA. Results: As expected, mice were severely hyperlipidemic upon high-fat feeding. In response to fibrate, cholesterol and triglyceride levels dropped dramatically. Phenotypic analysis of the liver showed macrophage infiltration already 2 days after high-fat feeding. Fat accumulation in the liver was noticed after 4 days followed by a severe steatosis at one week. Fibrate treatment decreased the inflammation in the liver and abolished the steatosis. Microarray analysis supported the observed rapid inflammation in response to high-fat feeding and the inhibition of this process by fibrate. In contrast, genes involved in lipid metabolism where regulated more gradually. Surprisingly, fibrate and the diet regulated lipid metabolism and inflammation through different sets of genes. Conclusions: The APOE2ki mouse can serve as an excellent model for steatohepatitis. These data indicate that inflammation might have a causal role in the development of steatosis.
I W02-P-01 3 1 T N F - a -308G>A P O L Y M O R P H I S M IS N O T A RISK FACTOR F O R CORONARY DISEASE (CHD) IN A GREEK-CYPRIOT COHORT 1 M. Tzirkalli , G. Dedoussis 1, C. Zambartas 2. 2Department of i
Dietetics-Nutrition, Harokopio University, Athens, Greece; 2Department of Cardiology, Nicosia General Hospital, Nicosla, Cyprus Objective: Tumor Necrosis Factor -alpha (TN'F-~) is a cytokine that has caused great scientific interest because of its involvement in most of the metabolic pathways, including the inflammatory process of atherosclerosis. The objective of the present study is to investigate the association of the occurrence of the TNF-a -308 G > A polymorphism with a predisposition to CHD. Methods: We studied demographic, lifestyle and clinical information of 56 non-hospitalized CHD patients and 51 healthy volunteers all of Greek Cypriot origin, matched by age and sex, without any clinical evidence of coronary heart disease. Genotyping was performed by PCR-RFLP analysis. P-values are two-sided and STATA 6 software was used for the calculations.
75th EAS Congress, 23-26 April 2005, Prague, Czech Republic
/ HERBAL TERPENOIDS ACT AS LIGANDS F O R PPAR-ALPHA AND G A M M A TO MANAGE GENE EXPRESSION INVOLVED IN LIPID METABOLISM AND I N F L A M M A T I O N
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 antihypercholesteremic 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 macrophages. 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 modifying 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 transferaselA 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 A L L E L E OF T H E C H O L E S T E R O L - 7 A L P H A HYDROXYLASE (CYP7A1) GENE DETERMINES HYPERRESPONSIVENESS OF L D L - C H O L E S T E R O L C O N C E N T R A T I O N 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 cholesterol 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 LDLcholesterol 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-OOSJSYNERGISTIC EFFECT OF STROMELYSIN-1 (MATRIX METALLOPROTEINASE-3) P R O M O T E R 5A/6A P O L Y M O R P H I S M W I T H S M O K I N G 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; 2lnsitute 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 A P O L I P O P R O T E I N E GENE P R O M O T E R -219G/T P O L Y M O R P H I S M DETERMINES INSULIN SENSITIVITY IN RESPONSE TO DIETARY FAT IN HEALTHY Y O U N G P E O P L E 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, independently 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 individuals. 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
Workshops W2 Genes and environment: nutrigenetics
10
IS when a MUFA- or a CHO-rich diets are consumed instead of a SFA-rich diet.
W02-P-010 I FOLATE LEVELS D E T E R M I N E E F F E C T OF ANTIOXIDANT SUPPLEMENTATION ON M I C R O N U C L E I IN SUBJECTS W I T H CARDIOVASCULAR RISK B. Smolkov~ ~, M. Du~insk~i 1, K. Rasqovfi I , M. Barancokov~i I , A. Kafimirov~ 1, A. Horskh 1, V. Spnstovh 1, J. Gagparovic 1, A. Collins 2.
1Research Base of Slovak Medical University, Institute of Preventive and Clinical Medicine, Bratislava, Slovakia; 2Department of Nutrition, University of Oslo, Norway
Objective: We studied the effect of modest supplementation with c~tocopherol (100 mg/day), ~-carotene (6 mg/day), vitamin C (100 mg/day) and selenium (50 ~tg/day) on oxidative stress and chromosomal damage markers. Subjects and methods: Two groups of middle aged men differing in cardiovascular risk [28 survivors of myocardial infarction before age 50 (MI), and 58 healthy controls (RC)] were randomly divided into equal groups to receive antioxidants or placebo for 12 weeks. Micronucleus (MN) levels in peripheral lymphocytes, and changes seen after intervention, were studied in relation to MTHFR C677Tgenotype, basal plasma homocysteine (hey) and relate levels. Ferric reducing ability of plasma (FRAP) and concentration of malondialdehyde (MDA) were measured to assess the antioxidant effect of supplementation. Results: MN frequency was not significantly associated with folate, hey or MDA levels before supplementation. MN frequencies and plasma folate levels did not vary significantly with MTHFR genotype. Hcy levels in subjects w i t h / T variant genotype were significantly higher compared with CT or CC (17.094-5.96 vs. 11.934- 4.34 and 12.224- 3.84 p.mol/L; P=0.001), especially in subjects with low folate (21.77±8.16 vs. 9.84± 2.41 and 14.584- 3.93 mol/L; P=0.012). In the placebo RC group (but not in the supplemented group) an increase of MN (from 5.064-3.59 to 7.284-5.47cells with MN; P=0.00) was detected at the end of the intervention period. In antioxidant-supplemented MI survivors we found an increase in FRAP (from 1000±163 to 1306±167 p,mol/L; P<0.001) and a decrease in MDA (from 2.344-1.0 to 1.334-0.4 Ixmol/L; P=0.001). MN frequency showed a decrease, which was strongest in subjects with normal folate levels (from 8.70 4- 4.71 to 4.054-2.29 cells with MN; P=0.015). In subjects with low folate, a high correlation was found between MN after supplementation and hey both before (r=- 0.979, P=0.002) and after supplementation (r=-0.922, P=0.009). Conclusions: Folate deficiency may amplify the effect of other risk factors such as elevated lacy levels or variant MTHFR genotype, as well as influencing the ability of antioxidant supplementation to protect against genetic damage. I W02-P-011 I M A R K E R S OF OXIDATIVE DAMAGE TO LIPIDS AND DNA RELATED TO SEASONAL VARIATIONS IN DIET I
B. Smolkov~ 1, M. Du~insk~ 1, K. Ra~lov~iI , G. McNeill 3, V. Spustov~ 1, P. Bla~cek 2 , A. Horsk~i I , J. Gagparovic 1, A. Conins 4. 1Institute of
Preventive and Clinical Medicine, Slovak Medical University, Bratislava, Slovakia; 2Hospital of Ministry of Defence, Bratislava, Slovakla; University of Aberdeen, Scotland, UK; 4Department of Nutrition, University of Oslo, Norway
Objective: The relatively high incidence of cardiovascular disease and cancer in countries of centraYeastem Europe might be associated with nutritional imbalance, particularly in winter months when traditionally the supply of fresh fruit and vegetables is limited. Subjects and methods: We monitored markers of oxidative stress and antioxidants in three Slovak population groups: 46 survivors of myocardial infarction (MI), 48 healthy, normolipidemic subjects (NL) and 70 rural controls (RC). Data were collected in February/March (winter/spring-Wl, W2) and September/October (summer/autumn-A1, A2) of two consecutive years, representing times of minimum and maximum local availability of fresh fruits and vegetables. Lipid peroxidation was assessed by the concentration of malondialdehyde (MDA) in plasma and lymphocyte DNA oxidation was measured using the comet assay. Dietary antioxidants, folate levels, homocysteine and total antioxidant status were measured in plasma.
Results: Food frequency questionnaires showed that vegetable consumption in summer/autumn was twice as high as in winter/spring (P<0.001). DNA damage did not vary consistently across the seasons, but mean plasma MDA levels for the MI and NL groups showed a clear pattern, with high levels in winter/spring and low levels in summer/autumn (/VII group: 3.604-1.98, 1.394-1.00, 1.894-1.03, 0.904-0.38 pmol/L; NL group: 1.85+0.90, 0.90+0.20, 1.27+0.66, 0.90±0.24 p,mol/L for W l , A1, W2, A2; P<10-7). Folate levels varied in the opposite way and followed the pattern of vegetable consumption (MI group: 4.20±1.3, 5.60±1.3, 3.964-1.4, 4.764-1.4 ng/ml; NL group: 4.624-1.5, 5.924-1.5, 5.054-2.7, 6.17±2.2 ng/ml for W l , A1, W2, A2; P<10-s). The RC group had the smallest seasonal variations - in vegetable consumption, folate levels, and MDA. Conclusions: High winter MDA levels are seen in those individuals with relatively low folate; they never occur in subjects with high plasma folate, implying that folate levels might directly protect against lipid oxidation. This study illustrates the value of the molecular epidemiological approach, while emphasising the need for well characterised population groups and valid biomarkers.
I WO2-P-012 I i
THE A P O E 2 K N O C K - I N M O U S E AS A M O D E L F O R TYPE III HYPERLIPIDEMIA AND STEATOHEPATITIS
R.S. Sverdlov-Shiri 1, K. Wouters 1, P.J.J. van Gorp 1, M.J.J. Gijbels 1, B. Staels 3, M. van Bilsen 2, M.H. Hofker 1. 1Dept. of Molecular Genetics,
University Maastrich~ Maastricht, The Netherlands; 2Dept. of Physiology, University Maastrich~ Maastricht, The Netherlands; 3Institute Paster,Lille, France The apolipoprotein E2 (APOE2) allele is the main cause of Type III hyperlipidemia in man. The APOE2 knock-in (APOE2ki) mouse develops severe diet-induced hyperlipidemia and is highly responsive to fibrate, a PPAR~ agonist. Objective: Obtaining a better understanding of the molecular and physiological mechanisms underlying hyperlipidemia Methods: APOE2ki mice were fed a high-fat diet or a high-fat diet in combination with fenofibrate. The mice were sacrificed at several time points after start of the diet. As a control, we used chow-fed APOE2ki mice. In addition to physiological measurements, microarray analysis was preformed using liver-derived mRNA. Results: As expected, mice were severely hyperlipidemic upon high-fat feeding. In response to fibrate, cholesterol and triglyceride levels dropped dramatically. Phenotypic analysis of the liver showed macrophage infiltration already 2 days after high-fat feeding. Fat accumulation in the liver was noticed after 4 days followed by a severe steatosis at one week. Fibrate treatment decreased the inflammation in the liver and abolished the steatosis. Microarray analysis supported the observed rapid inflammation in response to high-fat feeding and the inhibition of this process by fibrate. In contrast, genes involved in lipid metabolism where regulated more gradually. Surprisingly, fibrate and the diet regulated lipid metabolism and inflammation through different sets of genes. Conclusions: The APOE2ki mouse can serve as an excellent model for steatohepatitis. These data indicate that inflammation might have a causal role in the development of steatosis.
I W02-P-01 3 1 T N F - a -308G>A P O L Y M O R P H I S M IS N O T A RISK FACTOR F O R CORONARY DISEASE (CHD) IN A GREEK-CYPRIOT COHORT 1 M. Tzirkalli , G. Dedoussis 1, C. Zambartas 2. 2Department of i
Dietetics-Nutrition, Harokopio University, Athens, Greece; 2Department of Cardiology, Nicosia General Hospital, Nicosla, Cyprus Objective: Tumor Necrosis Factor -alpha (TN'F-~) is a cytokine that has caused great scientific interest because of its involvement in most of the metabolic pathways, including the inflammatory process of atherosclerosis. The objective of the present study is to investigate the association of the occurrence of the TNF-a -308 G > A polymorphism with a predisposition to CHD. Methods: We studied demographic, lifestyle and clinical information of 56 non-hospitalized CHD patients and 51 healthy volunteers all of Greek Cypriot origin, matched by age and sex, without any clinical evidence of coronary heart disease. Genotyping was performed by PCR-RFLP analysis. P-values are two-sided and STATA 6 software was used for the calculations.
75th EAS Congress, 23-26 April 2005, Prague, Czech Republic