- Email: [email protected]
Th-P16:417 Statins inhibit vascular valcification by restoring GAS6-AXL survival pathway in human aortic smooth muscle cells
Thurs&ty, June 22, 2006: Poster Session P17 Novel technologies for risk determhzation
IT h - P 1 6 : 4 1 5 1
I I
E F F I C A C Y A N D S A F E T Y O F FLUVASTATIN IN CHILDREN A N D A D O L E S C E N T S WITH HETEROZYGOUS FAMILIAL HYPERCHOLESTEROLEMIA
A. Van Der Graa£ 1 , M.C. Nierman I , J.C. Firth 2, K.H. Wolmaxans 2, A.D. MaxaJs 2, E. De Groot 1.1Departmentof Vascular Medicine, Academic
Medical Centre, Universi~ of Amsterdam, Amsterdam, The Netherlands: 2Departnwnt of Medicine, Groote Schuur Hospital arm Universi~ of Cape Town, Cape Town, South Aftqca Objectives: To assess whether early initiation of statin therapy for heterozygous familial hypercholesterolemia favourably aYfect lipid profiles or vascular morphological changes. Methods: Children and adolescents aged 10-16 years with heterozygous familial hypercholesterolemia were administered fluvastatin (80mg/day) for two years in a single-axm two-center study. Carotid B-mode-intima-media thickness (IMT) and M-mode arterial wall softness (V,) were recorded. Results: Eighty of the 85 enrolled subjects completed the trial. The median decrease in LDL-cholesterol from baseline at last study visit was 33.9%; median decreases in total cholesterol, triglycerides and apolipoprotein B were 27.1%, 5.3%, and 24.2%, respectively; the median increase in HDLcholesterol was 5.3%. Changes in carotid arterial wall thickness and softness versus baseline were fractional and statistically non-significant (delta IMT=0.005 m m [95% CI: -0.018 to 0.007 mm, n=83] and delta V,=0.017 [95% CI: -0.219 to 0.253, n=79]). Adverse events, all non-serious, were reported by 58 subjects (68.2%); four were suspected to be drug-related. Change in hormone levels and their sexual maturation were appropriate for this age group. Conclusions: Fluvastatin lowered LDL-cholesterol, total cholesterol and apolipoprotein B levels effectively over a prolonged period in children and adolescents with heterozygous familial hypercholesterolemia. Carotid IMT and wall stiffness remained largely unchanged. Funding: This study was funded by Novartis Pharma AG.
I T h - P 1 6 : 4 1 6 I P R O - O X I D A N T A C T I O N O F STATINS K. Tikhaze, Z. Lankin, I. Kaminniy, E. Arzamastseva, V. Kukharchuk.
Russian Cardiology Research Complex, Moscow, Russia Objectives: HMG-CoA reductase inhibitors (statins) axe known to be very effective lipid-lowering drugs they appear to inhibit not just cholesterol synthesis but also natural pro-antioxidant coenzyme Q, as well as the biosynthesis of Se-containing GSH-Px. On the contrary, some literature data refer to statins depressing enzyme-mediated superoxide generation which is considered as antioxidant action of this drug. Methods: Concentration of lipidhydroperoxides in LDL particles (peroxyLDL) was assessed by modified Fe3+-xylenol orange method. Results: In double-blind placebo-controlled trials we investigated peroxyLDL levels in plasma of patients with atherosclerosis during 6 mths administration of pravastatin and cerivastatin as well as after 2 mths of simvastatin administration. We have found that 6 mths treatment of patients with pravastatin (40 mg daily) or cerivastatin (0,4 mg daily) were followed by sufficient accumulation of peroxy-LDL in vivo (increase of baseline level by 30% and more than 4-fold, respectively). The level of peroxy-LDL after 2 mths treatment with simvastatin (20 mg daily) increased by about 2,5 times. GSH-Px activity in erythrocytes of patients following 6 mths pravastatin treatment decreased by 4,8 times. Patients with heart failure complicated with diabetes mellitus had significantly higher level of peroxy-LDL following statin administration as compared to those without statin treatment. Conclusions: HMG-CoA reductase inhibitors to have substantial gross pro-oxidant effect in vivo and one may suggest that coenzyme Q may be effective in the prevention of oxidative stress.
Methods: VSMC calcification was induced by inorganic phosphate (Pi) treatment and was evaluated with o-cresolphthalein complexone method and yon Kossa staining. Results: In HASMC, Pi induced calcium deposition in concentration (1.4-3.2 mM)- and time (1-10 days)-dependent manners. Pi also induced HASMC apoptosis, determined by histone revelation (ELISA), and activated caspase3, Bax, and Bad in concentration- and time-dependent manners. The caspase inhibitor, Z-VAD-FMK, inhibited Pi-induced H A S M C calcification and apoptosis, revealing the causal link between apoptosis and caldfication. Pi downregulated expression of Growth arrest-specific gene6 (Gas6), a vitamin K-dependent survival factor, and its receptor Axl, indicating that this pathway is important for Pi-induced apoptosis and calcification. Statins significantly inhibited Pi-induced HASMC calcification and apoptosis in a concentration-dependent manner. This was accompanied by abrogation of Pi-induced downregulation of Gas6, Axl, and subsequent reduction of Akt and Bad phosphorylation. Conclusion: Apoptosis is essential for Pi-induced HASMC calcification, where downregulation of Gas6-Axl and subsequent deactivation of Akt, activation of Bad axe an important mechanism. Statins inhibit H A S M C calcification through upregulation of Gas6-Axl survival pathway. Funding: This work was supported by grants from the Ministry of Health, Labor and Welfare, Japan.
P17
I
B.K. Son, K. Kozaki, K. Iijima, M. Eto, M. Akishita, Y. Senda, T. Nakano, Y. Ouchi. Dept. of Geriatric Medicbw, The Universi~ of Tokyo, Tokyo, Japan Objective: Vascular calcification is clinically important in the development of cardiovascular disease. In this study, we have tried to clarify the mechanism of vascular smooth muscle cells (VSMC) calcification and investigated the effect of statins, repoted to inhibit aortic valve calcification, on human aortic smooth muscle cells (HASMC) calcification.
NOVEL TECHNOLOGIES
FOR RISK
DETERMINATION
[ Th-P17:418 ] QUANTITATIVE TRAIT LOCI MAPPING OF METABOLIC SYNDROME SUSCEPTIBILITY GENES IN M O U S E S.W. Fouchier, C.J.A. Moen, K. Willems Van Dijk, R.R. Frants. Departnwnt
of Human Genetics, Leiden Universi~ Medical Center; Leiden, The Netherlands Objectives: The Metabolic Syndrome (MS) is chaxacterized by a broad and variable phenotype, caused by an interaction of complex genetic and environmental factors, such as diet and life-style. We set out to identify novel genes and pathways related to metabolic stress and early (reversible) stages of MS by integration of human and mouse genetics/genomics. Methods: To identify novel genes, we have performed a linkage analysis using a F2 mapping population between APOE3Leiden (C57BL/6J) and FVB mice and different dietary interventions; chow diet, low fat (LFC), Western type (W) and a high fat (HFW) diet. Furthermore, liver gene expression patterns of C57BL/6J, FVB and APOE3Leiden were analyzed using long oligo micro-arrays (20K) in a loop design. Results: Differences were found in body weight, lipid and glucose levels. FVB mice were the most sensitive to diet-induced changes: increased weight, total cholesterol and glucose levels and decreased triglycerides and free fatty acids, especially when fed the W diet. Quantitative trait loci, determining lipid levels in response to the different diets, were found on chr. 1, 3, 4, 5, 6, 7, 9, 16 and 17. The number of differentially expressed genes located in these QTLs were the highest when fed a chow diet (2620), followed by a HFW diet (846), a W diet (198) and the LFC diet (37). Many of these genes were involved in lipid metabolism, detoxification and the immune response. Conclusion: Identification of the dietary response genes located in the QTLs may provide a better understanding of pathways involved in the development of MS in humans. Funding: NGC.
Th-P17:419
I
I Th-P 16:4171 STATINS I N H I B I T V A S C U L A R C A L C I F I C A T I O N BY R E S T O R I N G G A S 6 - A X L SURVIVAL PATHWAY IN H U M A N A O R T I C S M O O T H M U S C L E CELLS
585
3
/ J
THE APOLIPOPROTEIN E GENE PROMOTER (-491A/T) P O L Y M O R P H I S M D E T E R M I N E S TRIACYLGLYCEROL PLASMA CONCENTRATION IN R E S P O N S E T O D I E T A R Y FAT
J.A. Moreno, F. P&ez-Jim~nez, P. P&ez-Maxt~nez, B. Cortes, M. Gaxcia-Salas, J. Ruano, A. Gallego, J. Ldpez-Miranda. Lipids and
Atherosclerosis Research Unit, Hospital Unh,ersitario Reina Sofia, C6rdoba, Spa#~ Objective: To determine whether the apoE gene promoter (-491A/T) polymorphism is related with significantly different lipid response to changes in the quantity and quality of dietary fat in healthy subjects.
XIV bztetTtational Symposium on Atherosclerosis, Rome, Italy, June 18-22, 2006
IT h - P 1 6 : 4 1 5 1
I I
E F F I C A C Y A N D S A F E T Y O F FLUVASTATIN IN CHILDREN A N D A D O L E S C E N T S WITH HETEROZYGOUS FAMILIAL HYPERCHOLESTEROLEMIA
A. Van Der Graa£ 1 , M.C. Nierman I , J.C. Firth 2, K.H. Wolmaxans 2, A.D. MaxaJs 2, E. De Groot 1.1Departmentof Vascular Medicine, Academic
Medical Centre, Universi~ of Amsterdam, Amsterdam, The Netherlands: 2Departnwnt of Medicine, Groote Schuur Hospital arm Universi~ of Cape Town, Cape Town, South Aftqca Objectives: To assess whether early initiation of statin therapy for heterozygous familial hypercholesterolemia favourably aYfect lipid profiles or vascular morphological changes. Methods: Children and adolescents aged 10-16 years with heterozygous familial hypercholesterolemia were administered fluvastatin (80mg/day) for two years in a single-axm two-center study. Carotid B-mode-intima-media thickness (IMT) and M-mode arterial wall softness (V,) were recorded. Results: Eighty of the 85 enrolled subjects completed the trial. The median decrease in LDL-cholesterol from baseline at last study visit was 33.9%; median decreases in total cholesterol, triglycerides and apolipoprotein B were 27.1%, 5.3%, and 24.2%, respectively; the median increase in HDLcholesterol was 5.3%. Changes in carotid arterial wall thickness and softness versus baseline were fractional and statistically non-significant (delta IMT=0.005 m m [95% CI: -0.018 to 0.007 mm, n=83] and delta V,=0.017 [95% CI: -0.219 to 0.253, n=79]). Adverse events, all non-serious, were reported by 58 subjects (68.2%); four were suspected to be drug-related. Change in hormone levels and their sexual maturation were appropriate for this age group. Conclusions: Fluvastatin lowered LDL-cholesterol, total cholesterol and apolipoprotein B levels effectively over a prolonged period in children and adolescents with heterozygous familial hypercholesterolemia. Carotid IMT and wall stiffness remained largely unchanged. Funding: This study was funded by Novartis Pharma AG.
I T h - P 1 6 : 4 1 6 I P R O - O X I D A N T A C T I O N O F STATINS K. Tikhaze, Z. Lankin, I. Kaminniy, E. Arzamastseva, V. Kukharchuk.
Russian Cardiology Research Complex, Moscow, Russia Objectives: HMG-CoA reductase inhibitors (statins) axe known to be very effective lipid-lowering drugs they appear to inhibit not just cholesterol synthesis but also natural pro-antioxidant coenzyme Q, as well as the biosynthesis of Se-containing GSH-Px. On the contrary, some literature data refer to statins depressing enzyme-mediated superoxide generation which is considered as antioxidant action of this drug. Methods: Concentration of lipidhydroperoxides in LDL particles (peroxyLDL) was assessed by modified Fe3+-xylenol orange method. Results: In double-blind placebo-controlled trials we investigated peroxyLDL levels in plasma of patients with atherosclerosis during 6 mths administration of pravastatin and cerivastatin as well as after 2 mths of simvastatin administration. We have found that 6 mths treatment of patients with pravastatin (40 mg daily) or cerivastatin (0,4 mg daily) were followed by sufficient accumulation of peroxy-LDL in vivo (increase of baseline level by 30% and more than 4-fold, respectively). The level of peroxy-LDL after 2 mths treatment with simvastatin (20 mg daily) increased by about 2,5 times. GSH-Px activity in erythrocytes of patients following 6 mths pravastatin treatment decreased by 4,8 times. Patients with heart failure complicated with diabetes mellitus had significantly higher level of peroxy-LDL following statin administration as compared to those without statin treatment. Conclusions: HMG-CoA reductase inhibitors to have substantial gross pro-oxidant effect in vivo and one may suggest that coenzyme Q may be effective in the prevention of oxidative stress.
Methods: VSMC calcification was induced by inorganic phosphate (Pi) treatment and was evaluated with o-cresolphthalein complexone method and yon Kossa staining. Results: In HASMC, Pi induced calcium deposition in concentration (1.4-3.2 mM)- and time (1-10 days)-dependent manners. Pi also induced HASMC apoptosis, determined by histone revelation (ELISA), and activated caspase3, Bax, and Bad in concentration- and time-dependent manners. The caspase inhibitor, Z-VAD-FMK, inhibited Pi-induced H A S M C calcification and apoptosis, revealing the causal link between apoptosis and caldfication. Pi downregulated expression of Growth arrest-specific gene6 (Gas6), a vitamin K-dependent survival factor, and its receptor Axl, indicating that this pathway is important for Pi-induced apoptosis and calcification. Statins significantly inhibited Pi-induced HASMC calcification and apoptosis in a concentration-dependent manner. This was accompanied by abrogation of Pi-induced downregulation of Gas6, Axl, and subsequent reduction of Akt and Bad phosphorylation. Conclusion: Apoptosis is essential for Pi-induced HASMC calcification, where downregulation of Gas6-Axl and subsequent deactivation of Akt, activation of Bad axe an important mechanism. Statins inhibit H A S M C calcification through upregulation of Gas6-Axl survival pathway. Funding: This work was supported by grants from the Ministry of Health, Labor and Welfare, Japan.
P17
I
B.K. Son, K. Kozaki, K. Iijima, M. Eto, M. Akishita, Y. Senda, T. Nakano, Y. Ouchi. Dept. of Geriatric Medicbw, The Universi~ of Tokyo, Tokyo, Japan Objective: Vascular calcification is clinically important in the development of cardiovascular disease. In this study, we have tried to clarify the mechanism of vascular smooth muscle cells (VSMC) calcification and investigated the effect of statins, repoted to inhibit aortic valve calcification, on human aortic smooth muscle cells (HASMC) calcification.
NOVEL TECHNOLOGIES
FOR RISK
DETERMINATION
[ Th-P17:418 ] QUANTITATIVE TRAIT LOCI MAPPING OF METABOLIC SYNDROME SUSCEPTIBILITY GENES IN M O U S E S.W. Fouchier, C.J.A. Moen, K. Willems Van Dijk, R.R. Frants. Departnwnt
of Human Genetics, Leiden Universi~ Medical Center; Leiden, The Netherlands Objectives: The Metabolic Syndrome (MS) is chaxacterized by a broad and variable phenotype, caused by an interaction of complex genetic and environmental factors, such as diet and life-style. We set out to identify novel genes and pathways related to metabolic stress and early (reversible) stages of MS by integration of human and mouse genetics/genomics. Methods: To identify novel genes, we have performed a linkage analysis using a F2 mapping population between APOE3Leiden (C57BL/6J) and FVB mice and different dietary interventions; chow diet, low fat (LFC), Western type (W) and a high fat (HFW) diet. Furthermore, liver gene expression patterns of C57BL/6J, FVB and APOE3Leiden were analyzed using long oligo micro-arrays (20K) in a loop design. Results: Differences were found in body weight, lipid and glucose levels. FVB mice were the most sensitive to diet-induced changes: increased weight, total cholesterol and glucose levels and decreased triglycerides and free fatty acids, especially when fed the W diet. Quantitative trait loci, determining lipid levels in response to the different diets, were found on chr. 1, 3, 4, 5, 6, 7, 9, 16 and 17. The number of differentially expressed genes located in these QTLs were the highest when fed a chow diet (2620), followed by a HFW diet (846), a W diet (198) and the LFC diet (37). Many of these genes were involved in lipid metabolism, detoxification and the immune response. Conclusion: Identification of the dietary response genes located in the QTLs may provide a better understanding of pathways involved in the development of MS in humans. Funding: NGC.
Th-P17:419
I
I Th-P 16:4171 STATINS I N H I B I T V A S C U L A R C A L C I F I C A T I O N BY R E S T O R I N G G A S 6 - A X L SURVIVAL PATHWAY IN H U M A N A O R T I C S M O O T H M U S C L E CELLS
585
3
/ J
THE APOLIPOPROTEIN E GENE PROMOTER (-491A/T) P O L Y M O R P H I S M D E T E R M I N E S TRIACYLGLYCEROL PLASMA CONCENTRATION IN R E S P O N S E T O D I E T A R Y FAT
J.A. Moreno, F. P&ez-Jim~nez, P. P&ez-Maxt~nez, B. Cortes, M. Gaxcia-Salas, J. Ruano, A. Gallego, J. Ldpez-Miranda. Lipids and
Atherosclerosis Research Unit, Hospital Unh,ersitario Reina Sofia, C6rdoba, Spa#~ Objective: To determine whether the apoE gene promoter (-491A/T) polymorphism is related with significantly different lipid response to changes in the quantity and quality of dietary fat in healthy subjects.
XIV bztetTtational Symposium on Atherosclerosis, Rome, Italy, June 18-22, 2006