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L 021 EVALUATION OF THE OXIDATIVE STRESS AND INFLAMMATORY MARKERS IN HYPERLIPIDEMIC PATIENTS
by circulating monocyte mitochondria to the oxidative stress frequently observed in high risk hyperlipidemic subjects. Twenty-nine primary hyperlipidemic subjects (14 hypercholesterolemic and 15 combined hyperlipidemic) and 18 normolipidemic individuals, without any drug treatment and non-smokers, were enrolled in this study. The mitochondrial monocyte ROS generation in intact polymorphonuclear cells was estimated by flow cytometry and plasma oxidized LDL (ox-LDL) was measured by ELISA. The hypercholesterolemic, but not the combined hypelipidemic subjects presented higher levels of monocyte ROS generation when compared to control group. Both hyperlipidemic subjects presented elevated plasma levels of total LDL, oxidized LDL and apolipoprotein B. Combined hyperlipidemic individuals presented additionally increased levels of small dense LDL particles, insulin, triglycerides, VLDL-cholesterol and free fatty acids. Furthermore, highly significants positives correlations between monocyte ROS generation with oxidized LDL and monocyte chemotactic protein 1 (MCP-1) levels, were found. These data suggest that mitochondrial ROS production by circulating monocytes from primary hyperlipidemic subjects can contribute to the oxidative stress and hence to atherosclerosis, disease frequently found in this group. The hyperlipidemia is also associated with high levels of inflammatory markers. A significant negative correlation between interleukin-6 (IL-6) and interferoninducible protein 10 (IP-10) levels found in hyperlipidemic individuals may suggest an antinflamatory propertie of citokine IL-6.
Methods and Results: The atherosclerotic lesion areas were determined by morphometric analysis of oil red O lipid-stained areas in the aortic root of male LDLR0 mice with ages between 9 to 70 weeks. The plasma lipids were determined using enzymatic kits (Rocheâ). Nine-week-old mice did not present any detectable atherosclerotic lesions in this aorta site. However, the 13-week-old mice presented lesion size of 4.7 ± 0.2 (mm2 x 103). Mice with 19, 24 and 70 weeks of age presented progressively larger lesions: 13.4 ± 4.4, 42.7 ± 8.5 e 655.5 ± 118.2 (mm2 x 103), respectively. There was a significantly positive correlation between lesion size and age (R= 0.91 and P < 0.0001) and the time-course of the lesion development fitted a exponential model (R = 0,86 and P < 0,0001). As expected, the plasma lipid concentrations also increased with the age. Conclusions: The LDLR0 mice develop atherosclerosis spontaneously and independently of unbalanced diets, which are detectable by lipid staining as early as the 13th week of life, and progress exponentially with ageing. Supported: Fapesp and CNPq. L 020
HETEROGENEITY AND MOLECULAR MECHANISMS OF THE ANTI-APOPTOTIC ACTIVITY OF HDL SUBFRACTIONS IN ENDOTHELIAL HUMAN CELLS
Juliana A Souza, Cecile Vindis, Sandrine Chantepie, Anne Negre-Salvayre, Patrice Therond, Robert Salvayre, Carlos V Serrano Junior, M John Chapman, Anatol Kontush
L 022
HC/FMUSP SÃO PAULO SP BRAZIL INSTITUTO DO CORAÇÃO and INSERM U. 551 PARIS FR France LDL and its oxydated forms have multiple atherogenic properties, including apoptosis induction of endothelial cells (HMEC). HDL has innumerable anti-atherogenic activities, such anti-oxidant, anti-inflammatory and antithrombotic and anti-apoptotic. The HDL subfractions (sHDL) are he terogeneous in their physical-chemical composition and biological activities. The anti-oxidant activity of sHDL increases with density and is deficient in patients with metabolic syndrome (SMet). However, the heterogeneity of the anti-apoptotic activity of sHDL is still unknown. The objectives of the study were: (i) to evaluate the heterogeneity of the protective activity of sHDL in normolipidemic individuals and in patients with SMet against HMEC apoptosis induced by LDLox; (ii) to define the molecular mechanisms involved in this activity. Through ultracentrifugation by density gradient, we have isolated five different sHDL. HMEC were incubated with LDLox, in the presence or not of sHDL. The markers of toxicity and of cellular apoptosis were evaluated. All sHDL protected HMEC against apoptosis induced by LDLox. The sHDL3c and 3b of normolipidemic individuals presented more potent antiapoptotic activity than the sHDL2a (p<0,01) and 2b (p<0,001). As well as the sHDL, they reduced the generation of ROS induced by LDLox, HDL3c being more potent than HDL2b (p<0.05). There was a positive correlation between the anti-apoptotic and intracellular antioxidant activities with content of apoA-I and sfingozin 1-phosphate (E1F) of the sHDL, seemingly dependent on the interaction with its receptor SR-BI. Finally, the isolated HDL3c (n=5) in patients with SMet have a significantly lower content of apoA-I and reduced anti-apoptotic activity (p<0,01), when compared to normolipidemics (n=5). There was a tendency for protection against the generation of ROS to decrease (SMet, n=10). We concluded that the sHDL3 powerfully protect the HMEC against apoptosis and the generation of ROS induced by LDLox, since the anti-apoptotic activity was reduced in the SMet. L 021
LIPID PROFILE DETERMINATION IN A GROUP OF PATIENTS FROM SOROCABA (SÃO PAULO/SP) CITY INFECTED BY HUMAN IMMUNODEFICIENCY VIRUS
Adriana Feitosa Peres, Heloísa Muller Destro, Lucia Nassi Castilho, Edilma Maria de Albuquerque Pharmacy Graduating Course, University of Sorocaba Sorocaba SP BRASIL e Clinical Pathology Department of Medical School/Unicamp, SP Campinas SP BRASIL The utilization of highly active antiretroviral therapy modified the prognostic of HIV-infected individuals. However, the increasing rates of survival expose them to complications not previously evidenced, such as increased coronary heart disease (CHD). The aim of the present study was to evaluate lipid profile in a group of patients infected by human immunodeficiency virus and the possible correlation between serum lipids levels and other laboratorial parameters (viral load, lymphocytes CD4 and CD8 counts). Until the moment, there are not published studies in Sorocaba (SP) evaluating lipid profiles of HIV-infected patients. We carried out a retrospective study of patients followed by Municipal Clinics of Sexually Transmitted Diseases (STD/AIDS), Sorocaba (SP/Brazil). The main inclusion criterion was a complete lipidic profile records. Four thousand laboratory records were analyzed and only 150 (3.8%) patients had complete lipidic profile measurement. Therefore, a total of 150 patients (57% male and 43% female) submitted to antiretroviral therapy (average time on drug treatment 5±1.9 y approximately), were included in this study. Forty-three percent of all HIV+ patients presented serum cholesterol levels ³200 mg/dL, 37% LDL-cholesterol ³100mg/dL, 24% HDL-cholesterol below 40 mg/dL and 59% triglycerides ³150 mg/dL. Serum cholesterol (p<0,0001) and triglycerides (p=0,0010) levels were positively correlated with the lymphocytes CD4 count. Glucose ³100mg/dL was observed in 21.3% of patients. We concluded that evaluation of lipid profile is not a common clinical practice utilized for screening and follow-up HIV-infected individuals. These patients presented two important risk factors for development for CHD: dyslipidemia concomitant with hyperglicemia.
EVALUATION OF THE OXIDATIVE STRESS AND INFLAMMATORY MARKERS IN HYPERLIPIDEMIC PATIENTS
L 023
Edilma Maria de Albuquerque, Rômulo Tadeu Dias de Oliveira, Larissa Sayuri Kato, Maria Heloísa Blotta, Anibal Eugênio Vercesi, Helena Coutinho Franco de Oliveira, Eliana Cotta de Faria, Lucia Nassi Castilho
A LATE BIMODAL POSTPRANDIAL LIPEMIA PATTERN IN WISTAR RATS DIFFERS FROM THAT IN MICE C57/BL6 AND HUMANS
Urban, A, Salerno, A G, Paim, B A, Oliveira, H C F, Faria, E C
Departament of Clinical Pathology/Faculty Medical Sciences Campinas SP BRASIL e Center of Medical and Experimental Surgery Campinas SP BRASIL
Universidade Estadual de Campinas Campinas SP BRASIL introduction: The pathophysiology of postprandial lipemia is not yet entirely clarified. TG, remnant-like particle cholesterol (RLP-col) and RLP- triglycerides increase after a fat load and could contribute to atherothrombosis. After a fat rich meal normolipidemic human beings present increased lipemia that lasts approximately 4 to 6h. Despite many studies showing postprandial lipe-
Atherosclerosis is characterized like an inflammatory chronic disease due to a high oxidative stress state generated by vascular wall cells. In present study, we investigated the contribution of reactive oxygen species (ROS) generation 22
Methods and Results: The atherosclerotic lesion areas were determined by morphometric analysis of oil red O lipid-stained areas in the aortic root of male LDLR0 mice with ages between 9 to 70 weeks. The plasma lipids were determined using enzymatic kits (Rocheâ). Nine-week-old mice did not present any detectable atherosclerotic lesions in this aorta site. However, the 13-week-old mice presented lesion size of 4.7 ± 0.2 (mm2 x 103). Mice with 19, 24 and 70 weeks of age presented progressively larger lesions: 13.4 ± 4.4, 42.7 ± 8.5 e 655.5 ± 118.2 (mm2 x 103), respectively. There was a significantly positive correlation between lesion size and age (R= 0.91 and P < 0.0001) and the time-course of the lesion development fitted a exponential model (R = 0,86 and P < 0,0001). As expected, the plasma lipid concentrations also increased with the age. Conclusions: The LDLR0 mice develop atherosclerosis spontaneously and independently of unbalanced diets, which are detectable by lipid staining as early as the 13th week of life, and progress exponentially with ageing. Supported: Fapesp and CNPq. L 020
HETEROGENEITY AND MOLECULAR MECHANISMS OF THE ANTI-APOPTOTIC ACTIVITY OF HDL SUBFRACTIONS IN ENDOTHELIAL HUMAN CELLS
Juliana A Souza, Cecile Vindis, Sandrine Chantepie, Anne Negre-Salvayre, Patrice Therond, Robert Salvayre, Carlos V Serrano Junior, M John Chapman, Anatol Kontush
L 022
HC/FMUSP SÃO PAULO SP BRAZIL INSTITUTO DO CORAÇÃO and INSERM U. 551 PARIS FR France LDL and its oxydated forms have multiple atherogenic properties, including apoptosis induction of endothelial cells (HMEC). HDL has innumerable anti-atherogenic activities, such anti-oxidant, anti-inflammatory and antithrombotic and anti-apoptotic. The HDL subfractions (sHDL) are he terogeneous in their physical-chemical composition and biological activities. The anti-oxidant activity of sHDL increases with density and is deficient in patients with metabolic syndrome (SMet). However, the heterogeneity of the anti-apoptotic activity of sHDL is still unknown. The objectives of the study were: (i) to evaluate the heterogeneity of the protective activity of sHDL in normolipidemic individuals and in patients with SMet against HMEC apoptosis induced by LDLox; (ii) to define the molecular mechanisms involved in this activity. Through ultracentrifugation by density gradient, we have isolated five different sHDL. HMEC were incubated with LDLox, in the presence or not of sHDL. The markers of toxicity and of cellular apoptosis were evaluated. All sHDL protected HMEC against apoptosis induced by LDLox. The sHDL3c and 3b of normolipidemic individuals presented more potent antiapoptotic activity than the sHDL2a (p<0,01) and 2b (p<0,001). As well as the sHDL, they reduced the generation of ROS induced by LDLox, HDL3c being more potent than HDL2b (p<0.05). There was a positive correlation between the anti-apoptotic and intracellular antioxidant activities with content of apoA-I and sfingozin 1-phosphate (E1F) of the sHDL, seemingly dependent on the interaction with its receptor SR-BI. Finally, the isolated HDL3c (n=5) in patients with SMet have a significantly lower content of apoA-I and reduced anti-apoptotic activity (p<0,01), when compared to normolipidemics (n=5). There was a tendency for protection against the generation of ROS to decrease (SMet, n=10). We concluded that the sHDL3 powerfully protect the HMEC against apoptosis and the generation of ROS induced by LDLox, since the anti-apoptotic activity was reduced in the SMet. L 021
LIPID PROFILE DETERMINATION IN A GROUP OF PATIENTS FROM SOROCABA (SÃO PAULO/SP) CITY INFECTED BY HUMAN IMMUNODEFICIENCY VIRUS
Adriana Feitosa Peres, Heloísa Muller Destro, Lucia Nassi Castilho, Edilma Maria de Albuquerque Pharmacy Graduating Course, University of Sorocaba Sorocaba SP BRASIL e Clinical Pathology Department of Medical School/Unicamp, SP Campinas SP BRASIL The utilization of highly active antiretroviral therapy modified the prognostic of HIV-infected individuals. However, the increasing rates of survival expose them to complications not previously evidenced, such as increased coronary heart disease (CHD). The aim of the present study was to evaluate lipid profile in a group of patients infected by human immunodeficiency virus and the possible correlation between serum lipids levels and other laboratorial parameters (viral load, lymphocytes CD4 and CD8 counts). Until the moment, there are not published studies in Sorocaba (SP) evaluating lipid profiles of HIV-infected patients. We carried out a retrospective study of patients followed by Municipal Clinics of Sexually Transmitted Diseases (STD/AIDS), Sorocaba (SP/Brazil). The main inclusion criterion was a complete lipidic profile records. Four thousand laboratory records were analyzed and only 150 (3.8%) patients had complete lipidic profile measurement. Therefore, a total of 150 patients (57% male and 43% female) submitted to antiretroviral therapy (average time on drug treatment 5±1.9 y approximately), were included in this study. Forty-three percent of all HIV+ patients presented serum cholesterol levels ³200 mg/dL, 37% LDL-cholesterol ³100mg/dL, 24% HDL-cholesterol below 40 mg/dL and 59% triglycerides ³150 mg/dL. Serum cholesterol (p<0,0001) and triglycerides (p=0,0010) levels were positively correlated with the lymphocytes CD4 count. Glucose ³100mg/dL was observed in 21.3% of patients. We concluded that evaluation of lipid profile is not a common clinical practice utilized for screening and follow-up HIV-infected individuals. These patients presented two important risk factors for development for CHD: dyslipidemia concomitant with hyperglicemia.
EVALUATION OF THE OXIDATIVE STRESS AND INFLAMMATORY MARKERS IN HYPERLIPIDEMIC PATIENTS
L 023
Edilma Maria de Albuquerque, Rômulo Tadeu Dias de Oliveira, Larissa Sayuri Kato, Maria Heloísa Blotta, Anibal Eugênio Vercesi, Helena Coutinho Franco de Oliveira, Eliana Cotta de Faria, Lucia Nassi Castilho
A LATE BIMODAL POSTPRANDIAL LIPEMIA PATTERN IN WISTAR RATS DIFFERS FROM THAT IN MICE C57/BL6 AND HUMANS
Urban, A, Salerno, A G, Paim, B A, Oliveira, H C F, Faria, E C
Departament of Clinical Pathology/Faculty Medical Sciences Campinas SP BRASIL e Center of Medical and Experimental Surgery Campinas SP BRASIL
Universidade Estadual de Campinas Campinas SP BRASIL introduction: The pathophysiology of postprandial lipemia is not yet entirely clarified. TG, remnant-like particle cholesterol (RLP-col) and RLP- triglycerides increase after a fat load and could contribute to atherothrombosis. After a fat rich meal normolipidemic human beings present increased lipemia that lasts approximately 4 to 6h. Despite many studies showing postprandial lipe-
Atherosclerosis is characterized like an inflammatory chronic disease due to a high oxidative stress state generated by vascular wall cells. In present study, we investigated the contribution of reactive oxygen species (ROS) generation 22