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YI-794 PARAOXONASE 1 PROMOTE HDL-MEDIATED CHOLESTEROL EFFLUX. IMPORTANCE OF PON1 FREE SULFHYDRYLS GROUP
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YI Young investigators
and larger in apoM-Tg (4.2±1.1%,p=0.06) compared with control mice (1.8±0.5%). Plasma PLTP activity was increased in apoM-Tg compared with control mice (16.0±0.95 versus 12.0±0.41 μmol/ml/hr, p<0.005), but was unchanged in apoM-/- mice. Plasma LCAT activity was not changed in apoM-Tg or apoM-/- mice. In conclusion, apoM affects conversion of α-HDL and formation of preβ-migrating HDL in plasma, at least ex vivo. This may affect the antiatherogenic function of HDL. YI-794
PARAOXONASE 1 PROMOTE HDL-MEDIATED CHOLESTEROL EFFLUX. IMPORTANCE OF PON1 FREE SULFHYDRYLS GROUP
H. Berrougui 1 , A. Khalil 2 . 1 Research Centre On Aging. Geriatric Service, Department of Medicine, University of Sherbrooke, Sherbrooke, Quebec, Canada It’s now well established that the anti-atherogenic properties of HDL (high density lipoproteins) are related to their capacity to protect LDL against lipid-peroxidation and to promote cholesterol efflux from macrophage. We have previously demonstrated that antioxidant activity of HDL decrease during ageing and it was attributed to the alteration of PON1. However, the role of PON1 in the cholesterol efflux process was until now poorly understood. The aim of this work was to investigate the mechanism of action by which PON1 stimulates cholesterol efflux. Our results show that purified PON1 from human plasma promotes in a dose dependent manner HDL mediated cholesterol efflux in J774 as well as in THP-1 macrophage. Furthermore, over expression of ABCA1-transporter J774 with cAMP analogue result in a significant increase of PON1 stimulating HDL mediated cholesterol efflux. We have also demonstrated that in the absence of HDL, PON1 was able to remove free cholesterol excess from macrophage in a dose dependent manner and that this effect was dependent of ABCA1 transporters. However, western blot analysis showed that incubation of macrophage with PON1 does not change ABCA1, SR-BI nor LXRα expression. Functional studies demonstrated that oxidation of PON1 by (o OH and Oo2 -oxygen free radicals produced by gamma-radiolysis of water) reduces dramatically the number of free sulfhydryls (–SH) and cholesterol efflux mediation. Our data show that PON1 plays a key role in the cholesterol efflux processes via an ABCA1 dependent pathway, and that this property was in major part dependent on the number of free –SH. YI-795
SMALL DENSE LDL PARTICLES AND METABOLIC SYNDROME IN A SAMPLE OF MIDDLE-AGED WOMEN FROM SOUTHERN ITALY. FINDINGS FROM PROGETTO ATENA
M. Gentile, S. Panico, F. Jossa, A. Mattiello, G. Marotta, P. Pauciullo, P. Rubba. Department of Clinical and Experimental Medicine, Federico II University, Naples, Italy Background: Metabolic Syndrome (MS) is highly prevalent in the general population. Small dense LDL (sd-LDL) particles have been considered as a possible risk marker in MS. Patients and Methods: We analyzed the relation between sd-LDL and MS in a population based sample of 210 middle-aged Southern Italian women; among them 86 participants had MS (40.9%). LDL particle separation was performed by Lipoprint System: 7 LDL subfractions were obtained, mean LDL particle size and LDL score (% of sd-LDL) were calculated. Results: Women with the MS had lower mean LDL particle size as compared to participants without MS (269 Å vs. 272 Å, p<0.001) but LDL score higher (3.6 vs. 0, p<0.001). The univariate analysis showed a significant association between MS and LDL score (upper quintile), (OR 4.80; 95% CI 2.29-10.18; p<0.001 for MS), apo B and insulin levels were positively related to the presence of sd-LDL. After controlling for age and insulin, MS remained related to high LDL score. After including apo B, MS was still related with LDL score (OR 4.0; 95% CI 1.76-9.09; p<0.001 for MS). Conclusion: Our results suggest that sd-LDL particles could be, in addition to other risk factors, a marker for diagnosis and severity of the MS. The LDL size measurement could be a tool for identification of patients at relatively high risk of cardiovascular disease within the large population with the MS, and who are candidate for intensive lipid-lowering interventions.
YI-796
COMPOSITIONAL AND METABOLIC ASSOCIATIONS OF LIPIDS IN VLDL, IDL, LDL, HDL2 AND HDL3 BY SELF-ORGANISING MAPS
L.S. Kumpula 1 , S.M. Mäkelä 2 , V-P. Mäkinen 1 , M.J. Savolainen 2 , K. Kaski 1 , M.L. Hannuksela 2 , M. Ala-Korpela 1 . 1 Laboratory of Computational Engineering, Systems Biology and Bioinformation Technology, Helsinki University of Technology, Helsinki, Finland; 2 Department of Internal Medicine, Clinical Research Centre, University of Oulu, Oulu, Finland The compositional variability within lipoprotein fractions and the metabolic relations between distinct particles are of fundamental interest. Analysis and visualisation of such multi-dimensional data is often hampered in conventional statistical tools. Here we introduce self-organising maps (SOMs) that transform multi-dimensional data into a two-dimensional map of individuals, where neighbours have similar lipid profiles. Each lipid variable can be visualised for the whole data set by colouring the map regions according to the average value of their local inhabitants. In this study VLDL, IDL, LDL, HDL2 and HDL3 particles were isolated and their lipid and protein compositions measured for 146 individuals. Twenty-five input parameters were used in the SOM analysis. An example of the results (for particle compositions per protein) is illustrated in the Figure. A group of individuals (upper boxes) with dominant VLDL1 (high TG and medium CE) also possess small TG-rich LDL and another group of individuals (lower boxes) with dominant VLDL2 (medium TG and high CE) have large CE-rich LDL with low TG.
These results, together with those that will be discussed in the presentation, demonstrate that SOM analyses reveal relevant compositional and metabolic associations in lipoprotein data and thus provide a new tool for lipoprotein research. YI-797
LECTIN-LIKE OXIDIZED-LDL RECEPTOR-1 (LOX-1) AND NITRIC OXIDE SINTHASE (NOS) POLYMORPHISMS INFLUENCES ATORVASTATIN ANTITHROMBOTIC ACTION AND CARDIOVASCULAR EVENT RATE
E. Capati 1 , F. Bruni 1 , A.L. Pasqui 1 , M. Pastorelli 1 , F. Ciani 2 , A. Palazzuoli 3 , A. Auteri 1 , L. Puccetti 1 . 1 Department of Internal Medicine and Immunology, Centre for Atherosclerosis Research, University of Siena, Siena, Italy; 2 Neurometabolic Unit, Meyer Children Hospital, Florence, Italy; 3 Department of Internal Medicine and Metabolic Diseases, Section of Cardiology, University of Siena, Siena, Italy Background and Aims: Recent data seem to indicate that statins offer CAD benefit even by mechanisms beyond lipid-lowering. Genetic influence has been shown for some statins anti-thrombotic actions via oxidized-LDL (ox-LDL) receptors and nitric oxide synthase (NOS) activity modulation (i.e. anti-platelet action). The aim of present study was to evaluate the cardiovascular (CAD) events rate according to ox-LDL lectin-like receptor-1 (LOX-1) and NOS polymorphisms distributionin in pure hypercholesterolemic subjects treated with atorvastatin. Methods: The present was a prospective four years study involving 1039 event-free subjects (643 male, 396 female) treated with atorvastatin (10-40 mg/day) to reach the estimated LDL target (3.36 mmol/L). Patients were evaluated every six months or at clinical event. LOX-1 3 UTR/T-C and NOS G894T polymorphisms were detected by allelic discrimination assays (PCR), lipid profile by enzymatic-colorimetric method, ox-LDL by ELISA, platelet activation by P-selectin expression (FACScan), NOS activity by intracellular citrullin recovery (iCit) (HPLC), C-reactive protein (CRP) by sensitive nephelometric technique, homocysteine by HPLC. Results: LOX-1 3 UTR/T (alone or in association with NOS G894T) was significantly associated to events in the whole cohort with respect to each other variable including LDL reduction (O.R. 4.90, 95% C.I. 3.196.98, p< 0.00001). Smoking influenced events in LDL-targeted subjects (p< 0.0001). Ox-LDL and P-selectin were better indicators than LDL or other variables according to 3 UTR/C genotype regardless magnitude of LDL reduction (O.R. 4.21, 95% C.I. 2.29-6.70 p< 0.0001).
76th Congress of the European Atherosclerosis Society, June 10–13, 2007, Helsinki, Finland
YI Young investigators
and larger in apoM-Tg (4.2±1.1%,p=0.06) compared with control mice (1.8±0.5%). Plasma PLTP activity was increased in apoM-Tg compared with control mice (16.0±0.95 versus 12.0±0.41 μmol/ml/hr, p<0.005), but was unchanged in apoM-/- mice. Plasma LCAT activity was not changed in apoM-Tg or apoM-/- mice. In conclusion, apoM affects conversion of α-HDL and formation of preβ-migrating HDL in plasma, at least ex vivo. This may affect the antiatherogenic function of HDL. YI-794
PARAOXONASE 1 PROMOTE HDL-MEDIATED CHOLESTEROL EFFLUX. IMPORTANCE OF PON1 FREE SULFHYDRYLS GROUP
H. Berrougui 1 , A. Khalil 2 . 1 Research Centre On Aging. Geriatric Service, Department of Medicine, University of Sherbrooke, Sherbrooke, Quebec, Canada It’s now well established that the anti-atherogenic properties of HDL (high density lipoproteins) are related to their capacity to protect LDL against lipid-peroxidation and to promote cholesterol efflux from macrophage. We have previously demonstrated that antioxidant activity of HDL decrease during ageing and it was attributed to the alteration of PON1. However, the role of PON1 in the cholesterol efflux process was until now poorly understood. The aim of this work was to investigate the mechanism of action by which PON1 stimulates cholesterol efflux. Our results show that purified PON1 from human plasma promotes in a dose dependent manner HDL mediated cholesterol efflux in J774 as well as in THP-1 macrophage. Furthermore, over expression of ABCA1-transporter J774 with cAMP analogue result in a significant increase of PON1 stimulating HDL mediated cholesterol efflux. We have also demonstrated that in the absence of HDL, PON1 was able to remove free cholesterol excess from macrophage in a dose dependent manner and that this effect was dependent of ABCA1 transporters. However, western blot analysis showed that incubation of macrophage with PON1 does not change ABCA1, SR-BI nor LXRα expression. Functional studies demonstrated that oxidation of PON1 by (o OH and Oo2 -oxygen free radicals produced by gamma-radiolysis of water) reduces dramatically the number of free sulfhydryls (–SH) and cholesterol efflux mediation. Our data show that PON1 plays a key role in the cholesterol efflux processes via an ABCA1 dependent pathway, and that this property was in major part dependent on the number of free –SH. YI-795
SMALL DENSE LDL PARTICLES AND METABOLIC SYNDROME IN A SAMPLE OF MIDDLE-AGED WOMEN FROM SOUTHERN ITALY. FINDINGS FROM PROGETTO ATENA
M. Gentile, S. Panico, F. Jossa, A. Mattiello, G. Marotta, P. Pauciullo, P. Rubba. Department of Clinical and Experimental Medicine, Federico II University, Naples, Italy Background: Metabolic Syndrome (MS) is highly prevalent in the general population. Small dense LDL (sd-LDL) particles have been considered as a possible risk marker in MS. Patients and Methods: We analyzed the relation between sd-LDL and MS in a population based sample of 210 middle-aged Southern Italian women; among them 86 participants had MS (40.9%). LDL particle separation was performed by Lipoprint System: 7 LDL subfractions were obtained, mean LDL particle size and LDL score (% of sd-LDL) were calculated. Results: Women with the MS had lower mean LDL particle size as compared to participants without MS (269 Å vs. 272 Å, p<0.001) but LDL score higher (3.6 vs. 0, p<0.001). The univariate analysis showed a significant association between MS and LDL score (upper quintile), (OR 4.80; 95% CI 2.29-10.18; p<0.001 for MS), apo B and insulin levels were positively related to the presence of sd-LDL. After controlling for age and insulin, MS remained related to high LDL score. After including apo B, MS was still related with LDL score (OR 4.0; 95% CI 1.76-9.09; p<0.001 for MS). Conclusion: Our results suggest that sd-LDL particles could be, in addition to other risk factors, a marker for diagnosis and severity of the MS. The LDL size measurement could be a tool for identification of patients at relatively high risk of cardiovascular disease within the large population with the MS, and who are candidate for intensive lipid-lowering interventions.
YI-796
COMPOSITIONAL AND METABOLIC ASSOCIATIONS OF LIPIDS IN VLDL, IDL, LDL, HDL2 AND HDL3 BY SELF-ORGANISING MAPS
L.S. Kumpula 1 , S.M. Mäkelä 2 , V-P. Mäkinen 1 , M.J. Savolainen 2 , K. Kaski 1 , M.L. Hannuksela 2 , M. Ala-Korpela 1 . 1 Laboratory of Computational Engineering, Systems Biology and Bioinformation Technology, Helsinki University of Technology, Helsinki, Finland; 2 Department of Internal Medicine, Clinical Research Centre, University of Oulu, Oulu, Finland The compositional variability within lipoprotein fractions and the metabolic relations between distinct particles are of fundamental interest. Analysis and visualisation of such multi-dimensional data is often hampered in conventional statistical tools. Here we introduce self-organising maps (SOMs) that transform multi-dimensional data into a two-dimensional map of individuals, where neighbours have similar lipid profiles. Each lipid variable can be visualised for the whole data set by colouring the map regions according to the average value of their local inhabitants. In this study VLDL, IDL, LDL, HDL2 and HDL3 particles were isolated and their lipid and protein compositions measured for 146 individuals. Twenty-five input parameters were used in the SOM analysis. An example of the results (for particle compositions per protein) is illustrated in the Figure. A group of individuals (upper boxes) with dominant VLDL1 (high TG and medium CE) also possess small TG-rich LDL and another group of individuals (lower boxes) with dominant VLDL2 (medium TG and high CE) have large CE-rich LDL with low TG.
These results, together with those that will be discussed in the presentation, demonstrate that SOM analyses reveal relevant compositional and metabolic associations in lipoprotein data and thus provide a new tool for lipoprotein research. YI-797
LECTIN-LIKE OXIDIZED-LDL RECEPTOR-1 (LOX-1) AND NITRIC OXIDE SINTHASE (NOS) POLYMORPHISMS INFLUENCES ATORVASTATIN ANTITHROMBOTIC ACTION AND CARDIOVASCULAR EVENT RATE
E. Capati 1 , F. Bruni 1 , A.L. Pasqui 1 , M. Pastorelli 1 , F. Ciani 2 , A. Palazzuoli 3 , A. Auteri 1 , L. Puccetti 1 . 1 Department of Internal Medicine and Immunology, Centre for Atherosclerosis Research, University of Siena, Siena, Italy; 2 Neurometabolic Unit, Meyer Children Hospital, Florence, Italy; 3 Department of Internal Medicine and Metabolic Diseases, Section of Cardiology, University of Siena, Siena, Italy Background and Aims: Recent data seem to indicate that statins offer CAD benefit even by mechanisms beyond lipid-lowering. Genetic influence has been shown for some statins anti-thrombotic actions via oxidized-LDL (ox-LDL) receptors and nitric oxide synthase (NOS) activity modulation (i.e. anti-platelet action). The aim of present study was to evaluate the cardiovascular (CAD) events rate according to ox-LDL lectin-like receptor-1 (LOX-1) and NOS polymorphisms distributionin in pure hypercholesterolemic subjects treated with atorvastatin. Methods: The present was a prospective four years study involving 1039 event-free subjects (643 male, 396 female) treated with atorvastatin (10-40 mg/day) to reach the estimated LDL target (3.36 mmol/L). Patients were evaluated every six months or at clinical event. LOX-1 3 UTR/T-C and NOS G894T polymorphisms were detected by allelic discrimination assays (PCR), lipid profile by enzymatic-colorimetric method, ox-LDL by ELISA, platelet activation by P-selectin expression (FACScan), NOS activity by intracellular citrullin recovery (iCit) (HPLC), C-reactive protein (CRP) by sensitive nephelometric technique, homocysteine by HPLC. Results: LOX-1 3 UTR/T (alone or in association with NOS G894T) was significantly associated to events in the whole cohort with respect to each other variable including LDL reduction (O.R. 4.90, 95% C.I. 3.196.98, p< 0.00001). Smoking influenced events in LDL-targeted subjects (p< 0.0001). Ox-LDL and P-selectin were better indicators than LDL or other variables according to 3 UTR/C genotype regardless magnitude of LDL reduction (O.R. 4.21, 95% C.I. 2.29-6.70 p< 0.0001).
76th Congress of the European Atherosclerosis Society, June 10–13, 2007, Helsinki, Finland