Efeito de um plano alimentar quali-quantitativo sobre a expressão gênica e concentração plasmática de biomarcadores inflamatórios em pacientes obesos em prevenção secundária para doença / Effect of qualitative and quantitative food plan on gene expression and plasma concentrations of inflammatory biomarkers in obese patients in secondary prevention for cardiovascular disease

Introdução: A doença cardiovascular é a maior causa de mortalidade no mundo e o Brasil está entre os 10 países com maiores índices, principalmente entre homens com idade inferior a 70 anos. A alimentação é um fator de risco modificável, que pode influenciar a expressão gênica e a concentração de biomarcadores inflamatórios relacionados à obesidade e à aterosclerose. Objetivo: Este subestudo avaliou se uma intervenção nutricional baseada na alimentação habitual brasileira modifica a expressão de genes envolvidos com aterosclerose e a concentração plasmática de biomarcadores inflamatórios, na prevenção secundária para doença cardiovascular. Métodos: Foram selecionados seis pacientes do sexo masculino, idade igual ou superior a 45 anos, obesos e com circunferência abdominal elevada, em acompanhamento do plano alimentar quali-quantitativo por 6 meses, para determinação da concentração plasmática de biomarcadores inflamatórios (interleucina (IL)-1, IL-6, IL-8, IL-10, IL-12, fator de necrose tumoral alfa, proteína C reativa e adiponectina) e expressão de 84 genes relacionados à aterosclerose em células totais do sangue periférico, bem como perfil lipídico, glicemia, insulinemia, ácidos graxos plasmáticos, dados sobre ingestão alimentar (recordatório de 24 horas) e antropometria (peso, estatura e circunferência da cintura), nas visitas inicial e final. Resultados: Após a intervenção nutricional, os pacientes reduziram o peso, a circunferência abdominal, o índice Homeostasis Model Assessment para resistência à insulina (p= 0,046) e a contagem global de leucócitos (p= 0,046) e de neutrófilos (p= 0,028). Não houve alteração significativa na concentração plasmática dos biomarcadores inflamatórios, contudo, verificou-se aumento significativo na expressão dos genes Apo A1 (p= 0,011), ELN (p= 0,017) e IL4 (p= 0,037). Conclusão: Assim, o plano alimentar quali-quantitativo, composto de alimentos habituais brasileiros, não reduziu a concentração de biomarcadores inflamatórios, mas aumentou a expressão de três genes envolvidos com aterosclerose, em pacientes obesos, na prevenção secundária para doença cardiovascular

Introduction: Cardiovascular disease is the largest cause of mortality in the world and Brazil is among the 10 countries with the highest rates, especially among men under 70 years of age. Diet is a modifiable risk factor, which may influence the gene expression and concentration of inflammatory biomarkers related to obesity and atherosclerosis. Objective: This substudy evaluated whether a nutritional intervention based on the usual Brazilian diet modifies the expression of genes involved with atherosclerosis and the plasma concentration of inflammatory biomarkers in the secondary prevention for cardiovascular disease. Methods: Six male patients, aged 45 years or older, obese and with high waist circumference were selected, to follow a qualitative-quantitative food plan for 6 months, in order to determine the plasma concentration of inflammatory biomarkers (interleukin (IL) -1), IL-6, IL-8, IL-10, IL-12, tumor necrosis factor alpha, C-reactive protein and adiponectin) and expression of 84 atherosclerosis-related genes in total peripheral blood cells, as well as lipid profile, glycemia, insulinemia, plasma fatty acids, food intake data (24-hour recall) and anthropometry (weight, height and waist circumference) at the initial and final visits. Results: After nutritional intervention, patients reduced weight, waist circumference, Homeostasis Model Assessment index for insulin resistance (p = 0.046) and overall leukocyte count (p = 0.046) and neutrophils (p = 0.028). There was no significant modification in the plasma concentration of the inflammatory biomarkers. However, there was a significant increase in the expression of Apo A1 (p = 0.011), ELN (p = 0.017) and IL4 (p = 0.037) genes. Conclusion: Thus, the qualitative-quantitative food plan, composed of Brazilian usual foods, did not reduce the concentration of inflammatory biomarkers, but increased the expression of three genes involved with atherosclerosis in obese patients, in secondary prevention for cardiovascular disease

Biochemical and genetic risk factors for atherosclerosis in systemic lupus erythematosus

Systemic lupus erythematosus [SLE] is associated with an increase in the risk of premature cardiovascular complications caused by accelerated atherosclerosis which significantly contributes to morbidity and mortality. Carotid ultrasonography is a very sensitive imaging tool to detect premature atherosclerosis and measurements of carotid intima-media thickness [IMT] assess the extent and the severity of systemic atherosclerosis. The pathogenesis of accelerated atherosclerosis in SLE is not clear; inflammation and endothelial dysfunction in addition to genetic risk factors represent important factors in the onset of atherosclerosis. To evaluate the relation between asymmetric dimethylarginine [ADMA], high sensitive C-reactive protein [hs-CRP], monocyte chemoattractant protein-1 [MCP-1] [both serum levels and the genotypes of the MCP-1 A-2518G polymorphism] with the development of carotid atherosclerosis in patients with SLE and their relation to disease activity. In the present study, 30 non-menopause SLE female patients and 20 healthy age-matched females were included. Both patients and controls were subjected to evaluation of body mass index [BMI], IMT, serum glucose, serum lipids, hs-CRP, ADMA, MCP-1 [both serum level and gene polymorphism]. Serum ADMA, hs-CRP, and MCP-1, levels were measured by enzyme-linked immunosorbent assay. MCP-1 genomic variants were detected by polymerase chain reaction followed by restriction enzyme-fragment analysis. Values for IMT, hs-CRP, ADMA and MCP-1 were significantly higher in patients with SLE than in healthy controls with more significant increase in SLE patients with IMT >/=1 mm than in those with IMT <1 mm. Carotid IMT was significantly positively correlated with all the studied variables except for age, BMI and FBS, but significantly negatively correlated with HDL-C in all SLE patients. G/G genotype of MCP-1 A-2518G gene was more frequent in SLE patients than controls. IMT, hs-CRP, ADMA and MCP-1 from patients with G/G phenotypes were markedly higher than those from patients with the A/A genotype. In multiple regression analysis, ADMA and MCP-1 were the strongest independent determinants of IMT in SLE patients. Assessment of high levels of ADMA, hs-CRP, MCP-1, in addition to the MCP-1 A-2518G polymorphism may play a role in the pathogenesis of accelerated atherosclerosis in SLE patients and would be useful in identifying the risk of developing cardiovascular disease. Development of a novel therapy targeting ADMA and MCP-1 may have a potential role in preventing the progression of increased IMT in SLE patients

Polimorfismo 2212A>G del gen PECAM-1 en individuos chilenos con enfermedad coronaria y controles / Effect of 2212A>G Polymorphism of PECAM-1 gene in Chilean subjects with coronary disease and controls

Introducción: La transmigración de leucocitos al espacio subendotelial es uno de los eventos claves en el desarrollo de la enfermedad arterial coronaria, siendo PECAM-1 (Platelet-endothelial cell adhesion molecule-1)una de las moléculas de adhesión responsables de este proceso. Objetivo: El objetivo de este trabajo fue evaluar la posible asociación entre el polimorfismo 2212A>G del gen PECAM-1 y enfermedad arterial coronaria. Pacientes y Método: Fueron evaluados 98 individuos con enfermedad coronaria confirmada angiográficamente (estenosis mayor al 70%) y 106 controles, por medio de la reacción en cadena de la polimerasa seguida de restricción enzimática (PCR-RFLP).Resultados: Los genotipos y frecuencias alélicas para el polimorfismo estudiado son similares en los dos grupos evaluados (p = 0.085 y p = 0.495 respectivamente). La OR relacionada al alelo mutado G fue 0.87 (I.C. 95%, 0.59 – 1.29, p = NS). Conclusión: El polimorfismo 2212A>G del gen PECAM-1 no se encuentra asociado con enfermedad arterial coronaria en individuos Chilenos.

Background: The transendothelial migration of leukocytes is a key event in coronary artery disease (CAD) development. Platelet endothelial cell adhesion molecule-1 (PECAM-1) is a cellular adhesion molecule responsible of this process. Aim: to investigate the possible association between 2212A>G polymorphism at the PECAM-1 gene and CAD in Chilean individuals. Methods: A total of 98 individuals whit CAD confirmed by angiography (>70% of stenosis) and 106 healthy controls were evaluated. The 2212A>G polymorphism at the PECAM-1 gene was analyzed by Polymerase Chain Reaction(PCR) and restriction fragment length polymorphism (RFLP). Results: The distribution of genotypes and relatives frequencies of alleles were similar between cases and controls (p = 0.085 and p = 0.495 respectively). The Odds Ratio of CAD whit the G allele was 0.87 (C.I: 95%, 0.59 –1.29;NS). Conclusion: 2212A>G polymorphism of the PECAM-1 gene was not associated whit CAD in Chilean individuals.

Polimorfismo C373G del gen PECAM-1 en individuos chilenos con enfermedad coronaria y controles / Polymorphism C373G of the PECAM-1 gene in chilean subjects with coronary disease and controls

The platelet endothelial cell adhesion molecule-1 (PECAM-1), a 130-kDa membrane glycoprotein, is expressed on the surface of monocytes, some T-lymphocyte subsets, neutrophils, platelets and endothelial cells. PECAM-1 plays a key role in the transendothelial migration of circulating leukocytes during vascular inflammation. The aim of the present investigation was to evaluate the association between the C373G polymorphism of the PECAM-1 gene and coronary artery disease in Chilean subjects. A total of 220 individuals were investigated (112 cases and 108 controls). The presence of coronary artery disease was confirmed by angiography (Stenosis >70 percent). The C373G polymorphism was detected by polymerase chain reaction followed enzymatic restriction. The genotype frequencies were in agreement with those predicted by the Hardy-Weinberg equilibrium in both groups. The genotype distribution and the relative alíele frequencies for C373G polymorphism of the PECAM-1 gene were similar between cases and controls (P= 0.820 and P= 0.739, respectively). Moreover, the OR associated with the mutated G alíele was 0.92 (C.I. 95 percent, 0.54 - 1.57; P=NS). In summary, our study showed that C373G polymorphism of the PECAM-1 gene is not associated with coronary artery disease in the population analyzed.

La molécula de adhesión celular endotelial plaquetaria-1 (PECAM-1) es una glicoproteína de membrana expresada por células endoteliales, plaquetas, monocitos, neutro filos y algunos tipos de linfocitos T. Constituye una pieza clave en la extravasación de leucocitos a través de las uniones intercelulares del endotelio vascular durante el proceso inflamatorio. El objetivo de nuestro estudio fue determinar la asociación entre el polimorfismo C373G (Leul25 Val) del gen PECAM-1 y enfermedad coronaria, en individuos chilenos. Se estudió un total de 220 individuos (112 casos y 108 controles). La presencia de enfermedad coronaria fue confirmada mediante angiografía (estenosis > 70 por ciento). El polimorfismo C373G, fue detectado mediante la técnica de reacción en cadena de polimerasa seguida de restricción enzimática. Las frecuencias genotípicas observadas en ambos grupos cumplen con la ley de Hardy-Weinberg. La distribución de genotipos como la frecuencia relativa de alelos para el polimorfismo C373G del gen PECAM-1, fueron similares entre casos y controles (p = 0.820 y p = 0.739, respectivamente). Además, la OR asociada al alelo mutado G fue 0.92 (I.C. 95 por ciento, 0.54-1.57; p= NS). Los datos obtenidos sugieren que el polimorfismo C373G del gen PECAM-1 no está asociado a enfermedad coronaria en la población analizada.

Farmacogenética e efeito antiinflamatório dos inibidores da HMG-CoA redutase / Pharmacogenetics and anti-inflammatory effect of HMG-CoA reductase inhibitors

A aterosclerose é resultado da associação de uma deposição de lipídios na parede arterial e um processo inflamatório de baixo grau. Essa inflamação pode ser detectada através da dosagem de marcadores séricos, que indicam o grau de aterosclerose, e estão associados a um maior risco de desenvolvimento de doenças cardiovasculares, independentemente dos níveis lipídicos. Entre estes marcadores destaca-se a Proteína C reativa ultra-sensível. As estatinas reduzem a inflamação associada à aterosclerose, o que é verificado por uma redução dos valores de proteína C reativa. Parte desse efeito está associada à diminuição de proteínas isopreniladas, porém as estatinas possuem efeitos diretos no sistema imune. Variações genéticas individuais estão associadas a variações no efeito hipolipemiante das estatinas, porém pouco se sabe sobre as variantes que interferem com as ações antiinflamatórias desses medicamentos. Além dos genes envolvidos no metabolismo do colesterol, genes que influenciam a farmacocinética e a farmacodinâmica das estatinas são possíveis responsáveis pela variação do efeito antiinflamatório observado.

Atherosclerosis is a result from the association of lipid deposition in the arterial wall and inflammatory process. This inflammatory process may be detected by clinical markers of systemic inflammation, such as ultrasensible C-reactive protein, which is associated with cardiovascular risk, independently of lipid levels. Statins reduce the inflammation associated to atherosclerosis, which may be verified by a reduction of the C-reactive protein levels. It seems that statins alter immune function by modulating post-translational protein prenylation. Individual genetic variations are associated with modulation of statins lipid-lowering effect; however, few studies have related the effect of the genetic variants with anti-inflammatory effect of statins. In addition to the genes involved in the cholesterol metabolism, genetic factors affecting statins pharmacodynamics and/or pharmacokinetics are potentially responsible for lipid and anti-inflammatory effects.

Hipercolesterolemia familiar / Familial hypercholesterolemia

A hipercolesterolemia familiar é uma doença genética, caracterizada por elevações dos níveis de colesterol plasmático, resultante da fração que não é removida adequadamente da circulação. São descritas mais de 600 mutações envolvidas nos mecanismos de síntese e expressão dos receptores da lipoproteína de baixa densidade (LDL), o que se traduz em redução ou em não funcionamento desses mecanismos. A forma de transmissão da mutação é autossômica dominante, o que resulta em dois fenótipos distintos: a forma homozigótica, rara, com prevalência de 1 em 1 milhão de indivíduos e valores de LDL-Colesterol acima de 600 mg/dl, além da presença de aterosclerose precoce, com acometimento cardiovascular já na primeira infância e adolescência; e a forma heterozigótica, mais frequente, que acomete 1 em 500 indivíduos, em que os níveis de LDL-colesterol plasmático se situam, geralmente entre 200 mg/dl e 400 mg/dl, e na ausência de tratamento adequado a doença coronariana vai se estabelecer em homens antes dos 50 anos e em mulheres antes dos 60 anos. o diagnóstico é estabelecido por meio de critérios clínicos e pode ser confirmado pela determinação da mutação. O tratamento, bem como as metas lipídicas a serem alcançadas, baseiam-se na estratificação de risco desses pacientes, o qual avalia, entre outros fatores, a presença de aterosclerose subclínica por meio da avaliação do complexo íntima média da carótida e do cálcio coronário. O diagnóstico de hipercolesterolemia familiar permite a identificação dessa doença em outros componentes assintmáticos em uma mesma família, podendo-se estabelecer o tratamento adequado da hipercolesterolemia, o que irá prevenir eventos cardiovasculares futuros

A doença arterial coronariana sob o prisma da genética molecular / A genetic view of the coronary artery disease

A biologia molecular evolui de forma vertiginosa e atualmente é tida como ferramenta indispensável na compreensão de doenças complexas e multifatoriais como a doença arterial coronariana. Tal abordagem gera uma nova forma de avaliação de doenças conhecidas e propicia a criação de novas técnicas, novos métodos diagnósticos e possíveis abordagens terapêuticas, interferindo diariamente no desfecho clínico final do paciente. O número de publicações em genética cardiovascular aumentou cinco vezes nos últimos 20 anos e a descoberta de novos polimorfismos e mutações bem como marcadores de inflamação, coagulação e genes relacionados ao metabolismo lipídico contribuem para se conhecer cada vez mais os aspectos intrínsecos envolvidos na aterosclerose. Este artigo irá abordar os principais avanços nessa área, identificando os polimorfismos mais comuns e sua relevância clínica segundo grandes ensaios e meta-análises,bem como fazer um breve racional acerca do desenho atual dos ensaios clínicos em biologia molecular

Apolipoprotein B gene polymorphisms in people in the East Mediterranean area of Turkey

Point mutations in the receptor binding domain of low density lipoprotein may increase cholesterol levels in blood. Three mutations of Apo B-100 protein result in defective binding [Arg 3500 ----> [corrected] Gln, Arg 3500 ----> [corrected] Trp and Arg 3531 ----> [corrected] Cys]. We estimated the frequency of Apo B point mutations [codon 3500] C9774T [Arg 3500 ----> [corrected] Trp] and G9775A [Arg 3500 ----> [corrected] Gln] in 179 atherosclerotic, 145 hyperlipidaemic individuals and 272 healthy individuals in the east Mediterranean region of Turkey. Lipid and lipoprotein levels were measured with routine biochemical analyser and Apo B mutation was detected using real-time PCR. Neither mutation was found. In this region, Apo B-100 protein mutations are rare and causes of hyperlipidaemia and atherosclerosis may therefore be unrelated to them

Endothelial cell oxidative stress and signal transduction

Endothelial dysfunction (ED) is an early event in atherosclerotic disease, preceding clinical manifestations and complications. Increased reactive oxygen species (ROS) have been implicated as important mechanisms that contribute to ED, and ROS's may function as intracellular messengers that modulate signaling pathways. Several intracellular signal events stimulated by ROS have been defined, including the identification of two members of the mitogen activated protein kinase family (ERK1/2 and big MAP kinase, BMK1), tyrosine kinases (Src and Syk) and different isoenzymes of PKC as redox-sensitive kinases. ROS regulation of signal transduction components include the modification in the activity of transcriptional factors such as NFkB and others that result in changes in gene expression and modifications in cellular responses. In order to understand the intracellular mechanisms induced by ROS in endothelial cells (EC), we are studying the response of human umbilical cord vein endothelial cells to increased ROS generation by different pro-atherogenic stimuli. Our results show that Homocysteine (Hcy) and oxidized LDL (oxLDL) enhance the activity and expression of oxidative stress markers, such as NFkB and heme oxygenase 1. These results suggest that these pro-atherogenic stimuli increase oxidative stress in EC, and thus explain the loss of endothelial function associated with the atherogenic process.

Actividad proliferativa y alteraciones cromosomicas de las ceculas musculares lisas en la ateroesclerosis / Proliferative activity and chromosomal alterations of smooth muscle cells in atherosclerosis

La causa de muerte más frecuente en los países desarrollados es la ateroesclerosis. Sus lesiones características, además del depósito lipídico, son el engrosamiento focal de la pared arterial con proliferación de células musculares lisas (CML) e infiltración mononuclear y presencia de vasos neoformados. En este trabajo estudiamos el fenómeno proliferativo y las alteraciones citogenéticas de las CML. Estas células, identificadas mediante inmunohistoquímica por su expresión de actina muscular específica, eran dipolides, con un alto índice de proliferación demostrado por expresión de la proteína nuclear PCNA. Un porcentaje elevado de CML expresó intensamente a la oconproteína p53. Además se encontraron claros indicios de inestabilidad cromosómica. Los hallazgos más frecuentes fueron trisomía del cromosoma 11. También se observó ampliación del gen FGF-3. Estos hallazgos permiten inferir que la proliferación de CML es activa, tiene relación con la acumulación o mutación de la oncoproteína p53 y además presenta alteraciones cromosómicas específicas y relacionadas con los factores de crecimiento. La presencia de este tipo de cambios nos lleva a considerar a la hiperplasia de las CML en la placa ateromatosa como una expresión celular de carácter clonal.

Diabete e aterosclerose: até onde a genética molecuilar pode nos levar? / Diabetes abd atherosclerosis: how far can molecular genetics lead us?

Recentes avanços na genética molecular de doença cardiovasculares vêm permitindo a possibilidade de identificaçäo de indivíduos com risco cardiovascular aumentado. O estudo de fatores de risco genético para diabete e aterosclerose näo só serve de modelo, mas coloca-se como ferramenta indispensável para a reduçäo da morbilidade e da mortalidade relacionadas a essas doenças durante a próxima década. Por meio da identidicaçäo de variantes genéticas que aumentem o risco individual e populacional, a identificaçäo pré-sintomática será possível, terapêuticas mais eficientes e individualizadas seräo utilizadas, e programas de prevençäo populacional contra o desenvolvimento dessas condições poderäp ser traçados de forma mais custo-efetiva.

Platelet aggregation and lipid profile in offsprings of young ischemics.

OBJECTIVE: To study the platelet aggregation and lipid profile in offsprings of young ischemic parents. DESIGN: Cross sectional. SETTING: Hospital based. METHODS: 40 adults under 45 years of age with proven CHD and their 50 offsprings comprised the study group. In the control group, 40 age matched adults with normal maximal treadmill test and their 50 off springs were included. All were screened for lipid profile and platelet aggregation. RESULTS: Platelet aggregation was significantly elevated in adults with CHD (44.0 +/- 15.5%) compared to controls (32.22 +/- 9.1%). Children of CHD adults also had significantly elevated aggregation (34.9 +/- 12.5%) compared to control offsprings (27.8 +/- 9.9%). Adult CHD subjects had significantly elevated serum cholesterol, LDL-cholesterol (LDL-C) and atherogenic index compared to control adults. The children in both groups had similar lipid profiles. CONCLUSION: Platelet aggregation is enhanced in offsprings of young ischemics. The potential of this index as a marker for early development of CHD needs to be explored.

Apolipoprotein E genotypes of normal and hyperlipidemic subjects

Apolipoprotein E (apo E) plays a role in the regulation of the lipid metabolism of humans. Apo E, 229 amino acid polypeptide, is classified into three major isoform (E2, E3, E4) according to the differences of amino acid in position 112 and 158. In the normal population apo E3 isoform is most prevalent and apo E2 or E4 is frequently associated with hyperlipoproteinemia. To find out the frequency of apo E isoform distribution in the Korean population, apo E genotyping was performed. After amplification of apoE gene by polymerase chain reaction (PCR), restriction isotyping was done by cleavage with restriction enzyme Hha I and polyacrylamide gel electrophoresis. The apo E allele frequency in 73 normal subjects was 4.8% for E2, 84.9% for E3 and 10.3% for E4. In diabetic patient with hyperlipoproteinemia, the frequency of apo E allele was 6.3% for E2, 81.0% for E3 and 12.7% for E4. There was no significant difference in apo E isoform distribution between diabetics and normal populations. But in patients with cardiovascular disease with hyperlipidemia, the apo E4 allele frequency was significantly higher than normal (20.0% vs 10.3%, p<0.005). Apo E3 was the most common isoform in normal and diabetic subjects and apo E2 isoform was rather low frequency compared to Caucasians. This pattern is similar to the Japanese population but somewhat different from other populations. From the data of a high association of apo E4 allele and cardiovascular disease with hypercholesterolemia, apo E isoform may be one of the determinants of hyperlipoproteinemia. The PCR method may be useful in apo E genotyping.