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W11.286 Atherogenic lipoprotein profile in rabbits after overexpression of human scavenger receptor class B type I
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Ragino T
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Workshops W12 Postprandial lipemia and atherosclerosis N E W M E T H O D F O R C O M P L E X EVALUATION OF S T R U C T U R A L C O M P O S I T I O N OF H D L S U B F R A C T I O N S B Y S M A L L - A N G L E X-RAY SCATTERING T E C H N I Q U E
Y. Ragino, F. Tusikov, M. Ivanova, E. Kashtanova, Y. Nikitin. Institute of Internal Medicine SB RAMS, Novosibirsk, Russia High density lipoproteins (HDL) comprise a heterogeneous group of pat: tides that differ in size, in density, in chemical composition and in theft" observed associations with coronary heat't disease (CHD). Of the five subfi'actions the three largest (HDL-2b, HDL-2a, HDL-3a) show the expected inverse correlation with CHD incidence and severity, whereas the two smallest subfi'actions (HDL-3b and HDL-3c) show a positive association. The new method for analysing of composition of 5 HDL subfi'actions was worked out based on the small-angle X-ray scattering (SAXS) technique and clinically approved in 119 patients with CAG documented CHD and 97 healthy subjects. The method allows qualitative and quantitative determinate the concentration and chemical composition of HDL subfi'actions (cholesterol, phospholipids and apo-LP contents in them) in serum. The results of HDL subfi'actions analysis by new method were compat'ed with gradient ultracentrifugation data. The results obtained by SAXS method when study the main lipid parameters (blood cholesterol, triglycerides and HDL-cholesterol) strong positive con'elated (r=0,91, r=0,87 and r=0,81, p<0,01, respectively) with those obtained by traditional biochemical enzymatic method. The new method for the determination of HDL subfi'actions composition is rapid (15 minutes), uses only one reagent (e.g., sucrose) and features a high accuracy and resolution up to HDL subfi'actions. A total of 0,05 ml of serum is requfl'ed for an assay. We believe that our method can be used in clinical practice for diagnostics, assessing and managing CHD risk in the general population.
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M A C R O P H A G E ACTIVATION, H Y P E R C O A G U L A B I L I T Y AND HYPOFIBRINOLYSIS. A L E T H A L TRIAD F O R PATIENTS W I T H L O W H D L - C H O L E S T E R O L LEVELS
O. S nchez-Sanchez, J. Mat'quez, E. Bernal, A. Bajo, A. Ugalde, D. Coca, M. Calbacho, E. Sanchez-Lat'go, R. Fabmgate, J. Sab n. Endothelial Pathology Unit., Ram n y Cajal Hospital, Madrid, Spain Introduction: Recently, low serum HDL levels is being considerated like a
independent factor in the endotelial dysfunction process. The novel mat'kers, like neoterin (Np), indicative of macrophage activation, allows us to know better the patogenia. Aims: - To determinate the prevalence of low and borderline HDL. - To correlate these with Np, vW factor (vWf) and PAI-1 M a t e r i a l and M e t h o d s : N: 177; 56 -4-13y, 89 M, 88 F, 112 hypertensive, 80 DM2, 40 smokers. Group A (high-normal HDL) N=136: HDL in M >=40, F>=50; B (borderline) N=19: M =35-39, F= 45-49; and C (low)N=22: M <35, F <45. BMI,waist circumference (W). Chol-t, HDL, triglycerydes: Hitachi auto-analyzer. LDL (Friedewald). PAI-1 (ng/ml); Np (ngr/ml) in N= 75 and vWf (U/ml) in N=77 by ELISA. Statistical analysis: T-Student. U-Mann Whitney Results: - The prevalence of low and borderline HDL was 12,2% and 11%, respectively. - Group C had BMI and W higher than A (p=0.000) and B (p=0.003) - T h e r e were statistically significant differences between PAI-1 levels in the different groups (A/C: p<0,002; B/C: p<0,005). - Np in B+C was lower than in A (p= 0,025). - In patients with Np >4,3, vW factor levels were higher than the patients with Np <=4,3 (p< 0,05) Conclusions: 1. There is a good correlation with BMI and W. 2. Our results suggest that patients with low HDL present, at least, macrophage activation, hypercoagulability and hypofibrinolysis, an explosive "cocktail" that could explain the high prevalence of cardiovasculat" events in these patients.
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I W l 1 . 2 8 6 1 I A T H E R O G E N I C L I P O P R O T E I N P R O F I L E IN R A B B I T S A F T E R OVEREXPRESSION OF H U M A N SCAVENGER R E C E P T O R CLASS B TYPE I I. Tancevski, S. Frank, P. Massoner, U. Stanzl, W. Schgoer, A. Wehinger, P. Eller, J. Patsch, A. Ritsch. Department of Medicine, University of Innsbruck, Innsbruck; Institute of Molecular Biology, Biochemistry and Microbiology, University of Graz, Graz, Austria B a c k g r o u n d : Scavenger receptor class B type I (SR-BI), a CD36 family
member, mediates selective uptake of HDL derived cholesterol. Crossing of LDL-receptor deficient mice with SR-BI transgene expressing mice led to decreased levels of HDL, LDL and VLDL-cholesterol and decreased atherosclerosis. In comparison to mice, rabbits do express cholesteryl ester transfer protein (CETP) in theft" plasma, show a more manlike lipoprotein profile and are susceptible to atherosclerosis. In this report we describe the effects of overexpression of human SR-BI in New Zealand White rabbits (NZW). M e t h o d s and ResuRs: 8 × 1011 adenoviral particles containing either hSR-BI o1" lacZ cDNA (control) were infused into the eat" vein of NZW rabbits. Lipoprotein parameters were determined fl'om blood samples drawn at days 0, 3, 7 and 10. Additionally, lipoprotein profiles were examined by FPLC analysis. Ad/hSR-BI infected rabbits showed a significant decrease in HDL-cholesterol with concomitant markedly increased levels of VLDL and LDL-cholesterol. Conclusion: Ad/hSR-BI overexpression in rabbits appealed to lead to an atherogenic lipoprotein profile by lowering HDL-cholesterol and increasing plasma VLDL and LDL levels. We feel that these data might help to further understand the role of SR-BI in atherogenesis.
I Wl 1.2871 N O R M O L I P I D E M I C
HEALTHY S U B J E C T S : ANALYSIS OF STABLE AND RADIOACTIVE ISOTOPE STUDIES
G. Watts, M. Farvid, S. Zilco, M. Allen, D. Chan, H. Ban'ett. School of Medicine and Pharmacology, University of Western Australia, Perth, Australia, and Tehran University of Medical Sciences, Tehran, Iran. The physiology of HDL apolipoprotein AI (apoA-I), a major player in reverse cholesterol transport and atherosclerosis, has not been fully elucidated. We examined the kinetic determinants of HDL apoA-I concentration using data fi'om 8 stable isotope (SIS) and 9 radioisotope (RIS) published studies. We selected normolipidemic, non-obese subjects with a BMI <29 kg/m 2, cholesterol <6, triglyceride (TG) <2.3 and HDL-cholesterol >0.9 retool/L; genetic hyperlipidemia and diabetes were excluded. We identified 43 subjects fi'om SIS and 45 fi'om RIS for analysis. There were no significant differences in age, BMI, gender, and plasma lipids between SIS and RIS subjects. The apoA-I fi'actional catabolic rate (FCR) was significantly higher in RIS compared with SIS (0.24-t-0.06 vs 0.21-t-0.05 d q , p=0.01); apoA-I production rate (PR) (12.34-3.16 vs 11.94-4.3 m g k g q d -1) was not different. In SIS, stepwise regression showed that apoA-I concentration was more significantly correlated with PR (r2=0.542, p<0.001) than FCR; HDL-cholesterol concentration was also significantly con'elated with PR (r2=0.157, p=0.009), but not FCR; apoA-I kinetics were not related to age, gender, TG o1" BML In RIS, apoA-I concentration was better con'elated with PR rather than FCR in stepwise regression analysis (r2=0.117, p=0.02); HDL-cholesterol concentration was negatively con'elated with FCR (r2=0.216, p=0.001). In conclusion, both SIS and RIS suggest that in normolipidemic subjects plasma HDL apoA-I is determined chiefly by PR, which in tm-n is dependent on genetic and envfl-onmental factors. This challenges the previous notion based on RIS and small number of selected subjects, suggesting that HDL-cholesterol concentrations are chiefly determined by HDL apoA-I catabolism.
74th EAS Congress, 17-20 April 2004, Seville, Spain
Ragino T
H
E
Workshops W12 Postprandial lipemia and atherosclerosis N E W M E T H O D F O R C O M P L E X EVALUATION OF S T R U C T U R A L C O M P O S I T I O N OF H D L S U B F R A C T I O N S B Y S M A L L - A N G L E X-RAY SCATTERING T E C H N I Q U E
Y. Ragino, F. Tusikov, M. Ivanova, E. Kashtanova, Y. Nikitin. Institute of Internal Medicine SB RAMS, Novosibirsk, Russia High density lipoproteins (HDL) comprise a heterogeneous group of pat: tides that differ in size, in density, in chemical composition and in theft" observed associations with coronary heat't disease (CHD). Of the five subfi'actions the three largest (HDL-2b, HDL-2a, HDL-3a) show the expected inverse correlation with CHD incidence and severity, whereas the two smallest subfi'actions (HDL-3b and HDL-3c) show a positive association. The new method for analysing of composition of 5 HDL subfi'actions was worked out based on the small-angle X-ray scattering (SAXS) technique and clinically approved in 119 patients with CAG documented CHD and 97 healthy subjects. The method allows qualitative and quantitative determinate the concentration and chemical composition of HDL subfi'actions (cholesterol, phospholipids and apo-LP contents in them) in serum. The results of HDL subfi'actions analysis by new method were compat'ed with gradient ultracentrifugation data. The results obtained by SAXS method when study the main lipid parameters (blood cholesterol, triglycerides and HDL-cholesterol) strong positive con'elated (r=0,91, r=0,87 and r=0,81, p<0,01, respectively) with those obtained by traditional biochemical enzymatic method. The new method for the determination of HDL subfi'actions composition is rapid (15 minutes), uses only one reagent (e.g., sucrose) and features a high accuracy and resolution up to HDL subfi'actions. A total of 0,05 ml of serum is requfl'ed for an assay. We believe that our method can be used in clinical practice for diagnostics, assessing and managing CHD risk in the general population.
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M A C R O P H A G E ACTIVATION, H Y P E R C O A G U L A B I L I T Y AND HYPOFIBRINOLYSIS. A L E T H A L TRIAD F O R PATIENTS W I T H L O W H D L - C H O L E S T E R O L LEVELS
O. S nchez-Sanchez, J. Mat'quez, E. Bernal, A. Bajo, A. Ugalde, D. Coca, M. Calbacho, E. Sanchez-Lat'go, R. Fabmgate, J. Sab n. Endothelial Pathology Unit., Ram n y Cajal Hospital, Madrid, Spain Introduction: Recently, low serum HDL levels is being considerated like a
independent factor in the endotelial dysfunction process. The novel mat'kers, like neoterin (Np), indicative of macrophage activation, allows us to know better the patogenia. Aims: - To determinate the prevalence of low and borderline HDL. - To correlate these with Np, vW factor (vWf) and PAI-1 M a t e r i a l and M e t h o d s : N: 177; 56 -4-13y, 89 M, 88 F, 112 hypertensive, 80 DM2, 40 smokers. Group A (high-normal HDL) N=136: HDL in M >=40, F>=50; B (borderline) N=19: M =35-39, F= 45-49; and C (low)N=22: M <35, F <45. BMI,waist circumference (W). Chol-t, HDL, triglycerydes: Hitachi auto-analyzer. LDL (Friedewald). PAI-1 (ng/ml); Np (ngr/ml) in N= 75 and vWf (U/ml) in N=77 by ELISA. Statistical analysis: T-Student. U-Mann Whitney Results: - The prevalence of low and borderline HDL was 12,2% and 11%, respectively. - Group C had BMI and W higher than A (p=0.000) and B (p=0.003) - T h e r e were statistically significant differences between PAI-1 levels in the different groups (A/C: p<0,002; B/C: p<0,005). - Np in B+C was lower than in A (p= 0,025). - In patients with Np >4,3, vW factor levels were higher than the patients with Np <=4,3 (p< 0,05) Conclusions: 1. There is a good correlation with BMI and W. 2. Our results suggest that patients with low HDL present, at least, macrophage activation, hypercoagulability and hypofibrinolysis, an explosive "cocktail" that could explain the high prevalence of cardiovasculat" events in these patients.
I
I W l 1 . 2 8 6 1 I A T H E R O G E N I C L I P O P R O T E I N P R O F I L E IN R A B B I T S A F T E R OVEREXPRESSION OF H U M A N SCAVENGER R E C E P T O R CLASS B TYPE I I. Tancevski, S. Frank, P. Massoner, U. Stanzl, W. Schgoer, A. Wehinger, P. Eller, J. Patsch, A. Ritsch. Department of Medicine, University of Innsbruck, Innsbruck; Institute of Molecular Biology, Biochemistry and Microbiology, University of Graz, Graz, Austria B a c k g r o u n d : Scavenger receptor class B type I (SR-BI), a CD36 family
member, mediates selective uptake of HDL derived cholesterol. Crossing of LDL-receptor deficient mice with SR-BI transgene expressing mice led to decreased levels of HDL, LDL and VLDL-cholesterol and decreased atherosclerosis. In comparison to mice, rabbits do express cholesteryl ester transfer protein (CETP) in theft" plasma, show a more manlike lipoprotein profile and are susceptible to atherosclerosis. In this report we describe the effects of overexpression of human SR-BI in New Zealand White rabbits (NZW). M e t h o d s and ResuRs: 8 × 1011 adenoviral particles containing either hSR-BI o1" lacZ cDNA (control) were infused into the eat" vein of NZW rabbits. Lipoprotein parameters were determined fl'om blood samples drawn at days 0, 3, 7 and 10. Additionally, lipoprotein profiles were examined by FPLC analysis. Ad/hSR-BI infected rabbits showed a significant decrease in HDL-cholesterol with concomitant markedly increased levels of VLDL and LDL-cholesterol. Conclusion: Ad/hSR-BI overexpression in rabbits appealed to lead to an atherogenic lipoprotein profile by lowering HDL-cholesterol and increasing plasma VLDL and LDL levels. We feel that these data might help to further understand the role of SR-BI in atherogenesis.
I Wl 1.2871 N O R M O L I P I D E M I C
HEALTHY S U B J E C T S : ANALYSIS OF STABLE AND RADIOACTIVE ISOTOPE STUDIES
G. Watts, M. Farvid, S. Zilco, M. Allen, D. Chan, H. Ban'ett. School of Medicine and Pharmacology, University of Western Australia, Perth, Australia, and Tehran University of Medical Sciences, Tehran, Iran. The physiology of HDL apolipoprotein AI (apoA-I), a major player in reverse cholesterol transport and atherosclerosis, has not been fully elucidated. We examined the kinetic determinants of HDL apoA-I concentration using data fi'om 8 stable isotope (SIS) and 9 radioisotope (RIS) published studies. We selected normolipidemic, non-obese subjects with a BMI <29 kg/m 2, cholesterol <6, triglyceride (TG) <2.3 and HDL-cholesterol >0.9 retool/L; genetic hyperlipidemia and diabetes were excluded. We identified 43 subjects fi'om SIS and 45 fi'om RIS for analysis. There were no significant differences in age, BMI, gender, and plasma lipids between SIS and RIS subjects. The apoA-I fi'actional catabolic rate (FCR) was significantly higher in RIS compared with SIS (0.24-t-0.06 vs 0.21-t-0.05 d q , p=0.01); apoA-I production rate (PR) (12.34-3.16 vs 11.94-4.3 m g k g q d -1) was not different. In SIS, stepwise regression showed that apoA-I concentration was more significantly correlated with PR (r2=0.542, p<0.001) than FCR; HDL-cholesterol concentration was also significantly con'elated with PR (r2=0.157, p=0.009), but not FCR; apoA-I kinetics were not related to age, gender, TG o1" BML In RIS, apoA-I concentration was better con'elated with PR rather than FCR in stepwise regression analysis (r2=0.117, p=0.02); HDL-cholesterol concentration was negatively con'elated with FCR (r2=0.216, p=0.001). In conclusion, both SIS and RIS suggest that in normolipidemic subjects plasma HDL apoA-I is determined chiefly by PR, which in tm-n is dependent on genetic and envfl-onmental factors. This challenges the previous notion based on RIS and small number of selected subjects, suggesting that HDL-cholesterol concentrations are chiefly determined by HDL apoA-I catabolism.
74th EAS Congress, 17-20 April 2004, Seville, Spain