Proefschrift Universiteit Maastricht. / Met bibliogr., lit. opg. - Met samenvatting in het Nederlands.

Proefschrift Universiteit Maastricht. / Met bibliogr. van de auteur, lit. opg. - Met samenvatting in het Nederlands.

Wall shear stresses are thought to have an influence on the formation of deposits of blood fats on the linings of the arteries, in atherosclerosis. Measuring velocities close to an artery wall to determine wall shears is difficult in view of the thinness of the boundary layer. Analytical solutions are limited to simple geometries and numerical analyses of three-dimensional, unsteady blood flows are expensive in terms of computational time. In the present study, finite element analyses of blood flow in models representative of the human aorta are based on two-dimensional sections in order to reduce the computational requirement.

OBJECTIVE: Atherosclerotic disease progression is mediated in part, by immunological mechanisms. In recent years, interest has increased towards the prospect of modulating these immune mechanisms through vaccination to ameliorate the course of disease. Patients with lupus are at a significantly higher risk for accelerated atherosclerosis and related complications. The goal of this study was to assess the outcome of immunization in mouse models of lupus, and lupus with accelerated atherosclerosis. MATERIALS/METHODS: Atherosclerosis-prone apoE^-/- mice and autoimmune gld mice were previously crossed to generate the gld.apoE^-/- mouse. Mice were treated with an apoB-100-derived vaccine, Alum (adjuvant control), or PBS control. The antibody response was determined by quantifying the amount of circulating anti-apoB100. Serum triglyceride and cholesterol levels were analyzed. Kidney tissue from gld and gld.apoE^-/- mice was processed and histologically analyzed, using glomerular tuft size as a measure of renal disease and by extension, autoimmune disease severity. Results: Immunization led to a pronounced initial antibody response that was decreased by the endpoint of the study. No significant differences in serum triglyceride or cholesterol were observed regardless of treatment. Similarly, no significant differences were observed in glomerular tuft size. Conclusion: The data suggests that immunization with an apoB-100- derived vaccine neither improves nor worsens autoimmune disease severity in the gld.apoE^-/- mouse model. It also appears that immunization is tolerated in the autoimmune background. While further study is necessary to determine the efficacy of immunization in reducing atherosclerotic disease in this model, this may be a possible therapy to lower incidence of atherosclerosis in lupus patients.

The pathogenesis of systemic lupus erythematosus (SLE) and cardiovascular disease (CVD) are tightly linked, and CVD is one of the leading causes of death in lupus patients. There are many risk factors that increase the risk of CVD in SLE patients, including endothelial dysfunction, lipid dysregulation, and abnormal regulation of innate and adaptive immunity. We have previously investigated the role of interferon regulatory factor 5 (Irf5), on atherosclerosis in lupus mouse models. Irf5 has a pro-inflammatory function by activating macrophage and cytokine recruitment and is thus being considered as a potential therapeutic target for the treatment of SLE. We hypothesized that Irf5 deficiency would ameliorate lupus disease as well as improve cardiovascular disease in the Irf5-deficient mouse model. However, while lupus disease did improve in the mouse model, the atherosclerotic plaques were found to be significantly increased in size. This poses a challenge to our current understanding of Irf5, as well as adds complexity to an already difficult clinical problem. Therefore, our aim of this study is to characterize the cells within the atherosclerotic lesions to examine their inflammatory potential. The focus of this study is the infiltration of macrophages into the mouse aortic root as determined by immunohistochemistry staining. In a time-course study using apoE.Irf5-/- mice, we found that macrophages started to accumulate into aortic leaflets as early as two weeks after starting a Western diet. Macrophage infiltration into the site of leaflet attachment seemed to possibly be a precursor to atherosclerotic lesion formation and appeared as early as 4 weeks after starting Western diet. No apparent differences were found between Irf5 sufficient and Irf5 deficient mice at either two or four weeks on Western diet. In a bone marrow chimera study, we examined the effects of Irf5 from bone marrow- and non-bone marrow-derived cells on the accumulation of macrophages on aortic leaflets and in the tunica intima in the gld.apoE-/- mouse model of lupus and atherosclerosis. Macrophage accumulation did not correlate with differences in Irf5 production. However, the finding of macrophage accumulation on aortic leaflets suggests a role of macrophages in Libman-Sacks endocarditis, an inflammatory disease of the mitral and aortic valves seen in patients with lupus. Together, our results do not support nor refute a role of Irf5 in macrophage infiltration into the aortic root. More samples are needed, as are more methods of identifying macrophages and quantifying them. However, it is still likely that macrophages play a role in the pathogenesis of atherosclerotic lesion formation in a lupus mouse model, and it is an area of study worth exploring. In a time-course study using apoE.Irf5-/- mice, we found that macrophages started to accumulate into aortic leaflets as early as two weeks after starting a Western diet. Macrophage infiltration into the site of leaflet attachment seemed to possibly be a precursor to atherosclerotic lesion formation and appeared as early as 4 weeks after starting Western diet. No apparent differences were found between Irf5 sufficient and Irf5 deficient mice at either two or four weeks on Western diet. In a bone marrow chimera study, we examined the effects of Irf5 from bone marrow- and non-bone marrow-derived cells on the accumulation of macrophages on aortic leaflets and in the tunica intima in the gld.apoE-/- mouse model of lupus and atherosclerosis. Macrophage accumulation did not correlate with differences in Irf5 production. However, the finding of macrophage accumulation on aortic leaflets suggests a role of macrophages in Libman-Sacks endocarditis, an inflammatory disease of the mitral and aortic valves seen in patients with lupus. Together, our results do not support a role of Irf5 in macrophage infiltration into the aortic root. However, it is still likely that macrophages play a role in the pathogenesis of atherosclerotic lesion formation in a lupus mouse model, and it is an area of study worth exploring.

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / Fundação de Amparo à Pesquisa do Estado do Rio de Janeiro / As células tronco são caracterizadas pela sua capacidade de se diferenciar em várias linhagens de células e exibir um pontente efeito parácrino. O objetivo deste trabalho foi avaliar o efeito da terapia com células da medula óssea (BMCs) na glicose sanguínea, no metabolismo lipídico e remodelamento da parede da aorta em um modelo experimental para aterosclerose. Camundongos C57BL/6 foram alimentados com uma dieta controle (grupo CO) ou uma dieta aterogênica (grupo AT - 60% gordura). Após 16 semanas, o grupo AT foi dividido em quatro sub grupos: grupo AT 14 dias e o grupo AT 21 dias receberam uma injeção de PBS na veia caudal e mortos 14 e 21 dias após respectivamente; grupo AT-BMC 14 dias e AT-BMC 21 dias que receberam uma injeção com BMCs na veia caudal e mortos 14 e 21 dias após, respectivamente. O grupo CO foi sacrificado juntamente com outros grupos. O transplante BMCs reduziu os niveis de glicose, triglicerídeos e colesterol total no sangue. Não houve diferença significativa em relação à massa corporal entre os grupos transplantados e não transplantados, sendo todos diferentes do grupo CO. Não houve diferença significativa na curva glicemica entre os grupos AT 14 dias, AT-BMC 14 dias e AT 21 dias e estes diferentes do grupo CO e do grupo AT-BMC 21 dias. O Qa (1/mm2) foi quantitativamente reduzido no grupo AT 14 dias e AT 21 dias quando comparado ao grupo CO. Este Qa se mostrou elevado no grupo AT-BMC 21 dias quando comparado a todos os grupos. O aumento da expessura da parede da aorta foi observado em todos os grupos aterogênicos, entretanto o aumento da espessura foi significativamente menor no grupo AT-BMC 21 dias em relação ao grupo AT 14 dias e AT 21 dias. A percentagem de fibras elásticas se apresentou significativamente maior no grupo AT 21 dias quando comparado ao CO e AT-BMC 21 dias. Não houve diferença significativa entre o grupo CO e AT-BMC 21 dias. Vacúolos na túnica média, delaminação e o adelgaçamento das lamelas elásticas foram observados nos grupos AT-14 dias e AT-21 dias. O menor número destes foi visualizado no grupo AT-BMC 14 dias e AT-BMC 21 dias. A imunomarcação para alfa actina de músculo liso (α-SMA) e fator de crescimento vascular e endotelial (VEGF) mostrou menor marcação em grupos transplantados com BMCs. A marcação para antígeno nuclear de proliferação celular (PCNA) mostrou-se mais expressiva no grupo AT-BMC 21 dias grupo. Marcação para CD105, CD133 e CD68 foi observada nos grupos AT 14 dias e AT 21 dias. Estas marcações não foram observadas nos grupos AT-BMC 14 dias e AT-BMC 21 dias. Nas eletromicrografias observamos o remodelamento benéfico no grupo AT-BMC14 dias e AT-BMC 21 dias, com a organização estrutural similar ao grupo CO. Vesículas de pinocitose, projeção da célula muscular lisa e a delaminação da lamina elástica interna são observados nos grupos AT 14 dias e AT 21 dias. Célula endotelial preservada, com lamina elástica interna de contorno regular e contínua é observada no grupo CO e nos grupos AT-BMC 14 dias e AT-BMC 21 dias. Como conclusão, os nossos resultados reforçam o conceito de que, em um modelo aterosclerótico utilizando camundongos e dieta aterogênica, a injeção de BMCs melhora os níveis de glicose, metabolismo lipídico e ocasiona um remodelamento benéfico na parede da aorta. / Stem cells are characterized by their ability to differentiate into multiple cell lineages and display the paracrine effect. The aim of this work was to evaluate the effect of therapy with bone marrow cells (BMCs) on blood glucose, lipid metabolism and aortic wall remodeling in mice through the administration of a high fat diet and subsequent BMCs transplantation. C57BL/6 mice were fed a control diet (CO group) or an atherogenic diet (AT group). After 16 weeks, the AT group was divided into four groups: an AT 14 days group and AT 21 days group, that were given an injection of vehicle and sacrificed at 14 and 21 days after, respectively; AT-BMC 14 days group and AT-BMC 21 days group that was given an injection of BMCs and sacrificed at 14 and 21 days after. The CO group was sacrificed along with other groups. The BMCs transplant had reduced blood glucose, triglycerides and total cholesterol. There was no significant difference in relation to body mass between the transplanted groups and non-transplanted groups, with all are different to CO group. There was no significant difference in the glycemic curve between AT 14 days group, AT-BMC 14 days group and AT 21 days group and these are different to CO and the AT-BMC 21 days group. The Qa (1 / mm2) was quantitatively reduced in the AT 14 days group and AT 21 days group when compared to the CO group. This Qa proved high in AT-BMC 21 days BMC compared to all groups. The increased thickness of the aortic wall was observed in all atherogenic groups, but was significantly smaller in group AT-BMC 21 days compared to AT 14 days group and AT 21 days group. The percentage of elastic fibers was significantly higher in the AT 21 days group when compared to the CO and AT-BMC 21 days. There was no significant difference between the CO and AT-BMC 21 days. Vacuoles in the media tunic, delamination and the thinning of the elastic lamellae were observed in AT 14 days group and AT 21 days group. The smallest number of these apresentation were displayed on the AT-BMC 14 days group and and AT-BMC 21 days. The immunostaining for α-SMA and VEGF showed lower in AT-BMC 14 days group and AT-BMC 21 days group. The markup for PCNA appears to be greater in the AT-BMC 21 days group. Marking to CD105, CD133 and CD68 were observed in AT 14 days group and AT 21 days group. These markings were not observed in AT-BMC 14 days group and AT-BMC 21 days group. In electron micrographs observed the beneficial remodeling in AT-BMC 14 day group and AT-BMC 21 days, with the structural organization was similar to the CO group. Vesicles of pinocytosis, projection of smooth muscle cell and delamination of the internal elastic lamina are seen in groups AT 14 days group and AT 21 days group. Endothelial cell preserved, regular and continuous contour in internal elastic lamelae is observed in the CO group, AT-BMC 14 days group and AT-BMC 21 days group. In conclusion, our results support the concept that an atherosclerotic model using mice and atherogenic diet, the injection of BMCs improve glucose, lipid metabolism and causes a beneficial remodeling of the aortic wall.

Orientador: Eliana Cotta de Faria / Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Ciencias Medicas / Made available in DSpace on 2018-08-09T21:29:39Z (GMT). No. of bitstreams: 1 Bassora_FernandaDutraSantiago_M.pdf: 2612060 bytes, checksum: 0ddcd23de6e1ce1f879247d7a9c9b2d7 (MD5) Previous issue date: 2007 / Resumo: Está bem estabelecida na literatura especializada a associação inversa entre as concentrações plasmáticas de colesterol das lipoproteínas de alta densidade (HDL-C) e a incidência de doença arterial coronariana (DAC). Além de propriedades anti-oxidante, anti-inflamatória e anti-trombótica, a HDL participa do transporte reverso de colesterol, via pela qual o colesterol é captado das lipoproteínas e das membranas células periféricas e transportado ao fígado para sua excreção na forma livre ou de ácidos biliares. A lipase hepática (LH) possui função crucial no transporte reverso do colesterol, por sua atividade lipolítica e pela função de ligante à lipoproteínas facilitando sua captação tissular. A proteína de transferência de ésteres de colesterol (CETP), e mesma importância metabólica, promove a troca de ésteres de colesterol por triglicérides entre a HDL e as lipoproteínas ricas em triglicérides. Mutações nos genes que codificam estas proteínas têm sido muito estudadas para se compreender a função destas no metabolismo lipídico. O modelo experimental da hiperalfalipoproteinemia tem sido utilizado no decorrer dos últimos anos com o intuito de elucidar os mecanismos de ação da HDL e das proteínas reguladoras do seu metabolismo. A hiperalfalipoproteinemia é caracterizada pelo aumento das concentrações de HDL-C e é causada principalmente por deficiências genética de CETP e/ou LH. Os objetivos desta dissertação foram o de se estabelecer à modulação da hiperalfalipoproteinemia sobre os parâmetros antropométricos, bioquímicos, moleculares (¿514C/T do gene da LH e I405V do gene da CETP) e radiológicos (espessura da camada íntima média de carótidas) em uma amostra populacional brasileira. O estudo foi conduzido em 291 voluntários de ambos os sexos, classificados como hiperalfalipoproteinemicos (Hiper-A), HDL-C =68mg/dL, ou controles, HDL-C<68 e =32 mg/dL, de acordo com o valor do percentil 90, obtido em um estudo prévio do Laboratório de Lípides a partir população normolipidêmica. Os polimorfismos LH-514C/T e CETP I405V foram identificados através de técnicas de reação em cadeia polimerase (PCR) e a espessura da camada íntima-média de carótidas (EIM) pela ultra-sonografia de alta resolução. Em um primeiro trabalho observou-se em um sub-grupo de 169 indivíduos, com a medida da EIM, que somente a idade foi correlacionada com a EIM na hiperalfalipoproteinemia, enquanto que em controles houve modulação positiva pela idade, sexo masculino, pressão arterial sistólica, e controversamente com relatos da literatura, com HDL-C. Apesar de Hiper-A possuir um perfil com maior número de fatores de risco cardiovasculares, a semelhança encontrada na EIM de carótidas, assim como, da freqüência de EIM maior que 1 mm poderia, em parte, ser explicada pela grande diferença de modulação entre os grupos apontando para um traço protetor contra a aterosclerose carotídea em hiperalfalipoproteinemia. A ateroproteção reduzida em controles, tanto em homens quanto em mulheres, está de acordo com a observada associação negativa neste grupo entre EIM e a CETP com possível presença de HDL com a composição química alterada (ricas em TG e pobres em ésteres de colesterol), e ocorreu possìvelmente no sub-grupo masculino, com perfil pró-aterogênico evidente. Em um segundo trabalho, no sub-grupo de 169 indivíduos, com a medida da EIM, foi avaliado o efeito do polimorfismo LH-514C/T sobre a espessura da camada íntima-média de carótidas na hiperalfalipoproteinemia. Não se observou nenhuma variação de EIM em ambos os grupos em função deste polimorfismo. Quando comparados os grupos, o genótipo CC do polimorfismo LH-514C/T mostrou apenas tendência a maior EIM de carótidas em hiperalfalipoproteinemia (p<0,09), mas a freqüência de EIM maior que 1 mm foi igual. Em um terceiro trabalho, em 282-291 indivíduos foram avaliadas as semelhanças de freqüências entre os polimorfismos LH-514C/T e CETP I405V na hiperalfalipoproteinemia e normolipidemia. Ambos apresentaram altas freqüências, similares entre grupos e entre o polimorfismo LH-514C/T, CC 39%, CT+TT 61%; e o polimorfismo CETP I405V: II 26%, IV+VV 74% e CTL: CC 40%, CT+TT 60%, II 43% e IV+VV 57%. Descrevemos o polimorfismo LH-514C/T na hiperalfalipoproteinemia os TT vs CC apresentaram cintura menor, concentrações mais baixas de colesterol plasmático (C), fosfolípides (FL), LDL-C, estimativa do tamanho da LDL (LDL-C/ApoB). O polimorfismo CETP I405V na hiperalfalipoproteinemia em VV vs II, mostrou alta pressão arterial sangüínea e menores concentrações plasmáticas de HDL2TG e HDL3TG. O genótipo IV teve maiores concentrações plasmáticas de ApoAI e pressão arterial diastólica quando comparado com o genótipo II. Em resumo esta dissertação aponta para efeitos ateroprotetores ou neutros da hiperalfalipoproteinemia em uma amostra de população brasileira sobre a aterosclerose carotídea, inclusive no polimorfismo LH -514C/T. Os polimorfismos LH-514C/T e CETP I405V foram muito semelhantes com relação aos lípides e lipoproteínas séricos, mas não às proteínas reguladoras, oferecendo modulação protetora na hiperalfalipoproteinemia / Abstract: There is an inverse relationship between plasma concentration of high-density lipoprotein (HDL-C) and the risk of coronary arterial disease (CAD). Beyond anti-oxidant, anti-inflammatory and anti-thrombotic properties, HDL plays a role on the reverse cholesterol transport, where cholesterol is taken from lipoproteins and peripheral cells to the liver for excretion. Hepatic lipase (HL) plays a key role in this process, by its lipolitic activities and ligand functions. Cholesterol ester transfer protein (CETP), of equal metabolic importance, facilitates the exchange of cholesterol ester and triglycerides between HDL and triglyceride rich-lipoproteins. Mutations and polymorphisms of these enzymes have being studied in order to evaluate its activity and metabolic consequences. Hyperalphalipoproteinemia (Hyper-A) has being used in the latest years with the purpose of evaluating the anti and pro-atherogenic mechanisms of HDL and of regulating proteins. The aim of this work was to establish the modulation of hyperalphalipoproteinemia in relation to controls on the anthropometric, biochemical, radiological and molecular manifestations. This study was conducted on 291 volunteers, classified as Hyper-A, HDL-C=68mg/dL and controls, HDL-C <68 e 32 mg/dL according to the percentile 90th, obtained from a local normolipidemic population study. We determined clinic data, lipid, lipoproteins and radiological parameters of volunteers. The HL-514C/T and CETP I405V polymorphism were determined by polymerase chain reaction methods. The carotid intima-media thickness measurements were performed high performance ultrasound. We showed in the first manuscript that although possessing a higher risk coronary vascular disease profile the similarity found in carotid could in part be explained by the striking differences in its modulation between the two groups, indicating a protective trait against carotid atherosclerosis in hyperalphalipoproteinemia. In the Hyper-A population, was only correlated with age, while in controls had a positive correlation with age, male sex, systolic blood pressure, and surprisingly with HDL-C. This dissociation between IMT and HDL-C could be accounted for by a small HDL particle number in CTL. In the manuscript 2, the ¿514C/T polymorphism did not contribute to variations in the carotid IMT and Hyper-A did not modulate the IMT variations, contrary to Rundek et al., (2002) who investigated the ¿514C/T polymorphism on variations in the carotid IMT in 87 stroke-free subjects suggested that CC genotypes had increase of carotid IMT, FMT and HALP. The HL-514C/T e CETP I405V polymorphisms, were no associate, were highly prevalent in the two groups but were not associated with HDL-C. In Hyper-A, LH-514C/T induced lower plasma cholesterol (C), phospholipids (PL), LDL-C and LDL size (LDL-C/ApoB). In Hyper-A CETP I405V decreased blood pressure, reduced TG in HDL subfractions 2 and 3 of (HDL2TG and HDL3TG) and increase ApoAI. The HL -514C/T polymorphism in Hyper-A the TT vs CC had lower waist hip-circumference, cholesterol (C) concentrations, phospholipids (PL), LDL-C and estimated size particle by LDL-C/ApoB. The genotype TT was different between 2 groups: in Hyper-A with relation the CTL, had lower HL, estimated size particle by TG/HDL-C and higher HDL2C, HDL3C, HDL3TG, ApoAI and C concentrations and had higher C, estimated size particle by LDL-C/ApoB, ApoAI, HDL2C, HDL3C and estimated size particle by TG/HDL-C. The CETP I405V polymorphism in Hyper-A, the VV vs II had higher Systolic Blood Pressure and lower HDL2TG e HDL3TG concentrations. The IV genotype had higher ApoAI concentration and Diastolic Blood Pressure. In Hyper A, the VV genotype had higher HDL2C, HDL3C, ApoAI, e TG concentrations and reduced concentration of VLDL- and estimated size particle of LDL by TG/HDL-C. In summary, this work indicates an athero-protector and neutral effect on the carotid atherosclerosis in Hyper-A between HL-514C/T and CETP I405V polymorphisms both modulated for plasma lipids more atheroprotective / Mestrado / Ciencias Basicas / Mestre em Clinica Medica

Orientador: Otavio Rizzi Coelho / Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Ciencias Medicas / Made available in DSpace on 2018-08-08T19:13:55Z (GMT). No. of bitstreams: 1 Tornatore_ThaisFranchini_M.pdf: 1907097 bytes, checksum: 5b1b181996681047fcf543481e55173d (MD5) Previous issue date: 2006 / Resumo: A aterosclerose é caracterizada pelo recrutamento de monócitos e linfócitos para a parede do vaso. O efeito desencadeador deste processo é o acúmulo de partículas de LDL minimamente oxidadas, as quais estimulam as células endoteliais a produzirem moléculas pró-inflamatórias. Em adição à LDL oxidada, vários outros fatores modulam a inflamação tais como forças hemodinâmicas, hormônios sexuais e infecção. O objetivo deste estudo foi demonstrar o efeito anti-aterogênico do DMA, 6,7-Dimetoxi-4-N-(3¿-N¿,N¿-dimetil)fenilaminoquinazolina, em camundongos LDLr-/- . Este efeito pode ser relacionado com a propriedade anti-inflamatória do composto estudado, por evidências de que o mesmo aumenta a biodisponibilidade de adenosina nos tecidos pela inibição da adenosina quinase. Nossos resultados mostraram que o DMA administrado via oral, tanto no tratamento preventivo, quanto no tratamento de lesões pré estabelecidas, em camundongos LDLr-/- alimentados com dieta hipercolesterolêmica (1,25% colesterol e 0,5% ácido cólico) diminui o tamanho das placas ateroscleróticas em 67% e 52%, respectivamente, em relação ao grupo controle que recebeu veículo. Esses dados foram obtidos através da planimetria das lesões ateroscleróticas na base da artéria aorta com a coloroção Oil Red O específica para lipídios. A dieta hipercolesterolêmica (HC) por 15 e 30 dias reduziu o triglicéride sérico, medido pelo sistema de cromatografia líquida, em cerca de 50% sendo que o tratamento com DMA cancelou esta redução. A dieta HC por 15 e 30 dias aumentou o colesterol sérico, em cerca de 2 vezes nos camundongos LDLr-/-. Camundongos tratados com DMA e dieta HC apresentaram níveis de colesterol total maiores do que o grupo alimentado somente com dieta HC. Dados revelaram que este aumento foi devido ao maior nível de HDL circulante nos camundongos tratados com o composto DMA. Todos os camundongos em tratamento, tanto com DMA quanto com veículo, apresentaram redução do peso corporal em relação ao grupo de camundongos alimentados com dieta hipercolesterolêmica (HC) / Abstract: Atherosclerosis is characterized by the recruitment of monocytes and lymphocytes to the artery wall. A triggering event for this process is the accumulation of minimmally oxidized LDL, which stimulates the overlying endothelial cells to produce a number of pro-inflammatory molecules, including hemodynamic forces, sex hormones and infection. We demonstrated that DMA, ,7-Dimethoxi-4-N-(3¿-N¿,N¿-dimethyl)phenilaminoquinazoline, have an anti-atherogenic effect in mice LDLr-/-. This effect could have a relationship with an anti-inflammatory property of adenosine in tissues, by the inhibition of adenosine kinase. Results showed that DMA administrated by oral via, as prevent treatment, as pre-established lesions, in mice LDLr-/- with hypercholesterolemic diet (1,25% cholesterol and 0,5% colic acid), decreases atherosclerotic lesions in 67% and 52% respectively in relation of the vehicle group. These data was obtained by planimmetric study of atherosclerotic lesions in the base of aortic arthery with Oil Red O especific for lipids. The hypercholesterolemic diet by 15 and 30 days, decreases serum tryglicerides in 50%, in the other way, the treatment with DMA cancelled this reduction. At the same time, HC diet by 15 and 30 days increases total cholesterol serum, in 2 times in mice LDLr-/-. Mice treatment with DMA and hypercholesterolemic diet showed serum cholesterol higher than the HC group (control). Results demonstrated that this increase have relationship with higher HDL serum level in mice treated with compound. All mice in treatment (DMA and Vehicle group) had the weight decreased. / Mestrado / Ciencias Basicas / Mestre em Clinica Medica

Orientador: Joyce Maria Annichino-Bizzacchi, Valder Roberval Arruda / Tese (doutorado) - Universidade Estadual de Campinas, Faculdade de Ciencias Medicas / Made available in DSpace on 2018-08-12T11:25:31Z (GMT). No. of bitstreams: 1 Fabri_DanielaRamos_D.pdf: 28613634 bytes, checksum: 506450d54c214e02520051f098953569 (MD5) Previous issue date: 2008 / Resumo: As complicacoes associadas a aterosclerose sao as causas mais comum de morte na populacao ocidental. O entendimento da aterosclerose como uma doenca inflamatoria e a associacao da coagulacao com fenomenos inflamatorios e complicacoes da doenca arterial oferecem novas oportunidades de prevencao e tratamento.Estudos populacionais demonstraram que o risco de infarto agudo do miocardio esta reduzido em 80% em homens hemofilicos A (HA) se comparados com controles pareados em idade e sexo. No entanto, lesoes ateroscleroticas iniciais foram identificadas em pequenos grupos de pacientes com coagulopatias por estudo ultrassonografico de arterias carotida e femural de forma semelhante aos controles pareados.O objetivo deste estudo e determinar o papel dos fatores VIII e IX dacoagulacao no desenvolvimento da doenca aterosclerotica em dois diferentesmodelos animais de dilipidemia. Comparamos grupos de animais deficientes deFVIII (HA) ou animais deficientes de FIX (HB) deficientes apoliproteina E (APOE-/-) ou deficientes de receptores de lipoproteina de baixa densidade (LDLR-/-) comanimais hemostaticamente normais deficientes apenas de apoE (APOE-/-) ou deLDLR (LDLR-/-). Todos animais da linhagem C57Bl/6, pareados em sexo e idade.Nossos resultados sugerem que o FVIII tem um papel protetor e tempodependente no desenvolvimento da aterosclerose nos animais deficientes de apoE, independente dos seus elevados niveis de colesterol. No entanto, essaprotecao nao foi evidente nos animais LDLR-/-.A deficiencia severa do FIX (<1%) nao protegeu contra o desenvolvimento da aterosclerose. Surpreendentemente, estes animais apresentaram significantemente mais lesoes nos periodos mais tardios nos dois modelos de doenca aterosclerotica utilizados. Os animais HB/APOE-/- apresentaram altos indices de mortalidade a partir de 20 semanas do periodo de dieta. Os elevados niveis de FIX nao influenciou o desenvolvimento da doenca arterial nos modelos apoE, mas mostrou-se como fator de risco para trombose venosa.Este estudo demonstra que o FVIII e o FIX tem papeis heterogeneos na evolução da aterosclerose, sugerindo participacao em mecanismos independentes ao da coagulacao no desenvolvimento da doenca arterial, e novos estudos devem ser realizados visando novas possibilidades terapeuticas tambem para os pacientes hemofilicos de idade mais avancada com risco de doenca cardiovasculares. / Abstract: Complications of atherosclerosis are the most common causes of death in Western societies. The knowledge that atherosclerosis is an inflammatory disease and coagulation affects the disease's complications offers new opportunities for prevention and treatment. Population studies demonstrated that the risk for myocardial infarction is reduced by 80% among hemophilia A (HA) men compared to age and gender-matched controls. However, early atherosclerotic lesions were readily identified in small cohorts of adults HA and hemophilia B (HB) by ultrasonography of carotid and femoral arteries in a similar fashion to an agecontrol male group. Here we sought to determine the role of FVIII and FIX on the development of atherosclerosis in two different mouse models. We compared a group of FVIII (HA) or FIX deficient mice (HB) lacking the low-density lipoprotein receptor (LDLR- /-) or apolipoprotein E (APOE-/-) with hemostatically normal littermate controls lacking either LDLR (LDLR-/-) or apoE (APOE-/-). All mice were on C57Bl/6 background and all groups were matched for gender and age. Our results suggest that FVIII has a protective and time dependent effect on development of atherosclerosis in apoE deficient mice, independently of higher cholesterol levels. Notably, FVIII deficiency did not influence the vascular disease of LDLR-/- mice. Severe FIX deficiency (<1% of normal) did not protect against the development of atherosclerosis. Unexpectedly, the rates of lesions were higher at late time points in the APOE-/- model. In addition, high levels of FIX was did not influence the vascular disease in APOE-/- whereas venous thrombosis was documented in 3/6 mice. Moreover, in the HB/APOE-/- model, the rates of atherosclerosis were higher at week 22 and beyond this age these mice presented high rates of mortality than controls. These data demonstrate that FVIII and FIX play a rather heterogeneous role in the development of atherosclerosis which suggest that there are coagulation independent mechanisms in the development of vascular diseases. Further studies in hemophilia B subjects and carriers are warranted to define the FIX effect on the onset and progression of occlusive vascular disease which may raises concerns on the onset cardiovascular risks on an aging hemophilia population. / Doutorado / Doutor em Clínica Médica

Abstract Patients with chronic renal insufficiency (CRI), also at its early stages, have an elevated risk of cardiovascular disease (CVD). Many well-established risk factors of CVD co-occur in CRI, e.g. dyslipidemia and hypertension. The present studies investigated the association between renal function and selected CVD risk factors. The fractional catabolic rate of low-density lipoprotein apolipoprotein B (LDL FCR) has previously been found to be reduced in patients with severe CRI or on dialysis. This study investigated the LDL FCR in 57 patients with moderate to severe CRI and not on dialysis. Although the mean LDL FCR was comparable between the CRI patients and healthy controls, among the renal patients the LDL FCR was correlated with renal function, whereas it was significantly reduced only in patients with advanced CRI (estimated glomerular filtration rate &lt;15 mL/min/1.73 m2). Leptin is a protein regulating food intake and energy expenditure and it is also involved in lipid metabolism. Hyperleptinaemia is a known feature of CRI patients; they are thought to be leptin resistant. The association between leptin and the lipoprotein profile was studied in 73 CRI patients with moderate to severe CRI and not on dialysis. Leptin was associated with lipid and lipoprotein concentrations in the renal patients, as in the control subjects, pointing towards a poorer lipoprotein profile with higher leptin levels. Hypertensive subjects in whom nocturnal blood pressure (BP) declines by less than 10% (non-dippers) show more organ damage than those in whom it falls by more than 10% (dippers). Here, non-dipping was found in 19% of middle-aged subjects (226 males, 234 females) evaluated with ambulatory BP monitoring. The non-dippers had significantly lower renal function as compared with the dippers, and dipping status was a significant predictor of the variation in eGFR. Furthermore, an increased risk of non-dipping was observed among subjects with only minor decreases in renal function. Carotid intima-media thickness (cIMT) can be used as a surrogate marker of early atherosclerosis. The present study investigated the association between renal function and cIMT in middle-aged subjects (247 males, 258 females). Renal function was independently associated with cIMT among males and also among postmenopausal women. The increased cIMT was seen in conjunction with mild renal impairment. In conclusion, the catabolism of LDL correlated with the renal function among CRI patients, but it was significantly reduced only in patients with advanced CRI. Leptin concentrations correlated with the lipoprotein profile in CRI patients. Among general middle-aged subjects, even a mild decrease in renal function was associated with derangements in BP regulation and with increased carotid atherosclerosis.