Bax inhibitor 1 inhibits vascular calcification in mice by activating optic atrophy 1 expression / 南方医科大学学报

OBJECTIVE@#To investigate the effects of Bax inhibitor 1 (BI- 1) and optic atrophy protein 1 (OPA1) on vascular calcification (VC).@*METHODS@#Mouse models of VC were established in ApoE-deficient (ApoE-/-) diabetic mice by high-fat diet feeding for 12 weeks followed by intraperitoneal injections with Nε-carboxymethyl-lysine for 16 weeks. ApoE-/- mice (control group), ApoE-/- diabetic mice (VC group), ApoE-/- diabetic mice with BI-1 overexpression (VC + BI-1TG group), and ApoE-/- diabetic mice with BI-1 overexpression and OPA1 knockout (VC+BI-1TG+OPA1-/- group) were obtained for examination of the degree of aortic calcification using von Kossa staining. The changes in calcium content in the aorta were analyzed using ELISA. The expressions of Runt-related transcription factor 2 (RUNX2) and bone morphogenetic protein 2 (BMP-2) were detected using immunohistochemistry, and the expression of cleaved caspase-3 was determined using Western blotting. Cultured mouse aortic smooth muscle cells were treated with 10 mmol/L β-glycerophosphate for 14 days to induce calcification, and the changes in BI-1 and OPA1 protein expressions were examined using Western blotting and cell apoptosis was detected using TUNEL staining.@*RESULTS@#ApoE-/- mice with VC showed significantly decreased expressions of BI-1 and OPA1 proteins in the aorta (P=0.0044) with obviously increased calcium deposition and expressions of RUNX2, BMP-2 and cleaved caspase-3 (P= 0.0041). Overexpression of BI-1 significantly promoted OPA1 protein expression and reduced calcium deposition and expressions of RUNX2, BMP-2 and cleaved caspase-3 (P=0.0006). OPA1 knockdown significantly increased calcium deposition and expressions of RUNX2, BMP-2 and cleaved caspase-3 in the aorta (P=0.0007).@*CONCLUSION@#BI-1 inhibits VC possibly by promoting the expression of OPA1, reducing calcium deposition and inhibiting osteogenic differentiation and apoptosis of the vascular smooth muscle cells.

Biomarcadores y marcadores de imagen de la enfermedad de Alzheimer / Biomarkers and image markers of Alzheimer's disease

Este artículo presenta un análisis desde el punto de vista bibliográfico de marcadores y biomarcadores de la enfermedad de Alzheimer (EA). Las metodologías usadas fueron los marcadores de imágenes (Resonancia Magnética y Tomografía por emisión de positrones) y biomarcadores de la proteína BA42, de la proteína Tau y de la Apoliproteína E (ALPE). De esta manera, son de importancia los niveles de BA42 disminuidos, la Tau incrementada, los polimorfismos de ALPE y las alteraciones constatadas en los marcadores de imagen, como factores de riesgo esenciales para el desarrollo de la EA. Se realiza una revisión de la literatura con respecto a los hallazgos clínicos de esta enfermedad.

This article presents a bibliographical analysis of markers and biomarkers of Alzheimer's disease (AD). The methodologies used were the imaging markers (Magnetic Resonance and Positron Emission Tomography) and biomarkers of the BA42 protein, Tau protein and Apoliprotein E (ALPE). Thus, decreased levels of BA42, increased Tau, ALPE polymorphisms, and alterations in imaging markers are important as risk factors for the development of AD. A review of the literature is made regarding the clinical findings of this disease.

Antiinflamatorios no esteroides: seguridad gastrointestinal, cardiovascular y renal / Nonsteroidal antiinflammatory drugs: gastrointestinal and cardiovascular and renal safety

La selección de un medicamento específico perteneciente a una clase farmacológica es bajo criterios de eficacia, seguridad, costo y conveniencia. Los Antiinflamatorios No Esteroideos (AINEs) actualmente se constituyen en uno de los medicamentos más consumidos en el mundo, por lo tanto es de gran importancia la revisión de los aspectos de seguridad de este grupo farmacológico. El presente trabajo tiene el objetivo de analizar bajo las evidencias disponibles hasta la actualidad, la seguridad de los AINES con 3 criterios principales: gastrolesividad, cardiotoxicidad y nefrotoxicidad.

The choice of a specific medication belonging to a drug class is under the criteria of efficacy, safety, cost and suitability. NSAIDs currently constitute one of the most consumed drugs in the world, so it is very important review of the safety aspects of this drug class. This review has the objective of analyze the safety of NSAIDs on 3 main criteria: gastrolesivity, cardiotoxicity and nephrotoxicity.

Efeito dos resíduos de café seco e fermentado por Monascus ruber no metabolismo de camundongos Apo E / Effect of dry coffee residues fermented with Monascus ruber on the metabolism of Apo E mice

FUNDAMENTO: A aterosclerose é uma doença inflamatória crônica de origem multifatorial que ocorre em resposta à agressão endotelial. O fungo Monascus ruber apresenta atividade hipocolesterolêmica e polifenóis presentes no resíduo de café apresentam atividade antioxidante, podendo auxiliar na prevenção de doenças cardiovasculares. O resíduo de café possui quantidade significativa de açúcares fermentescíveis, constituindo-se em substrato apropriado para o cultivo de fungos. OBJETIVO: O objetivo deste estudo foi avaliar o efeito dos resíduos de café seco e fermentado por Monascus ruber no metabolismo lipídico de camundongos knockout Apo E. MÉTODOS: O ensaio biológico foi realizado com 30 camundongos knockout para o gene Apo E, divididos em cinco grupos e submetidos a diferentes tratamentos. Foi realizada a prospecção fitoquímica e quantificação de compostos fenólicos dos resíduos fermentado e sem fermentar. O soro dos animais foi analisado utilizando kits enzimáticos e o tecido aórtico incluso em parafina e corado com H/E para realização da análise histopatológica. RESULTADOS: O resíduo de café sem fermentar 2%, em relação ao grupo controle, diminuiu em 42% o nível sérico de triacilgliceróis e em aproximadamente 41% a fração VLDL-c. Os grupos dos animais alimentados com 10% de resíduo não fermentado e 2% de resíduo fermentado diminuíram a área de lesão 10,5% e 15,4%, respectivamente, quando comparados ao controle. O resíduo fermentado apresentou um teor de compostos fenólicos superior ao resíduo não fermentado. CONCLUSÃO: O presente estudo mostra que a fermentação do resíduo de café apresenta potencial efeito benéfico sobre as doenças cardiovasculares, especialmente a aterosclerose.

BACKGROUND: Atherosclerosis is a chronic inflammatory disease of multifactorial origin, which occurs in response to endothelial injury. The fungus Monascus ruber has hypocholesterolemic activity, and the polyphenols present in coffee residue have an antioxidant activity and can help prevent cardiovascular diseases. Coffee residue has a significant amount of fermentable sugars, being an adequate substrate for growing fungi. OBJECTIVE: The objective of this study was to assess the effect of dry coffee residue fermented with Monascus ruber on the lipid metabolism of ApoE knockout mice. METHODS: The biological assay was performed with 30 ApoE knockout mice, divided into five groups and undergoing different treatments. The phytochemical prospection and quantification of phenolic compounds of the fermented and non-fermented coffee residues were performed. The sera of the animals were analyzed by using enzyme kits, and the aortic tissue was embedded in paraffin and stained with hematoxylin and eosin to undergo histopathological analysis. RESULTS: Comparing with the control group, the group receiving 2% non-fermented coffee residue showed a reduction of 42% in the serum levels of triacylglycerols and of approximately 41% in VLDL-c. The groups receiving 10% non-fermented coffee residue and 2% fermented coffee residue showed reductions in the lesion areas of 10.5% and 15.4%, respectively, as compared with the control group. The fermented coffee residue showed a higher content of phenolic compounds as compared with the non-fermented coffee residue. CONCLUSION: The present study showed that coffee residue fermentation has a potentially beneficial effect on cardiovascular diseases, especially atherosclerosis.

Grape extract and α-Tocopherol effect in cardiovascular disease model of Apo E -/- Mice / Efeito do extrato de uva e α-Tocoferol em camundongos Apo E -/-, modelo de doença cardiovascular

PURPOSE: To verify the effect of consumption of grape extract isolated or combined with α-tocopherol supplementation on atherosclerosis model with Apo E -/- mice. METHODS: After six weeks of atherogenic diet, Apo E -/- mice were divided into the following groups: Control, Grape, Tocopherol and Grape plus Tocopherol. The treatment progressed for 11 weeks when animals were submitted to euthanasia. RESULTS: All the treatments presented hypocholesterolemic effect with reduction of serum and liver cholesterol levels. This effect was parallel to an increase in the fecal excretion of cholesterol. There was also a higher fecal excretion of saturated fatty acids in groups receiving grape extract or α-tocopherol. All the groups treated presented a tendency to show higher levels of vitamin E. The fatty acid profile showed a tendency for monounsaturated fatty acid preservation after grape extract and α-tocopherol consumption. Morphological analysis revealed a lower degree of evolution of the atherosclerotic plaque of the animals that were fed α-tocopherol combined with grape extract, even when no difference was found in the size of the largest lesion. CONCLUSION: A synergistic effect between the polyphenols and α-tocopherol was observed, resulting in diminished evolution of atherosclerosis and a greater beneficial effect on atherosclerosis than the isolated consumption of antioxidants.

OBJETIVO: Verificar o efeito do consumo de extrato de uva isolada ou combinada com a suplementação de α-tocoferol em modelo de aterosclerose, utilizando camundongos Apo E -/-. MÉTODOS: Os camundongos Apo E -/- foram tratados com dieta aterogênica por seis semanas e foram divididos em quatro grupos: Controle, Uva, Tocoferol e Uva e Tocoferol. Após 11 semanas de tratamento os animais foram submetidos à eutanasia. RESULTADOS: Todos os tratamentos apresentaram efeito hipocolesterolêmico, com redução de colesterol plasmático e hepático. Este efeito foi acompanhado de um aumento na excreção fecal de colesterol. Houve também uma maior excreção fecal de ácidos graxos saturados nos grupos que receberam extrato de uva ou de α-tocoferol. Todos os grupos apresentaram uma tendência a apresentar níveis mais elevados de vitamina E. O perfil de ácidos graxos mostrou uma tendência para a preservação de ácidos graxos monoinsaturados, após consumo de extrato de uva e α-tocoferol. A análise morfológica revelou um menor grau de evolução da placa aterosclerótica dos animais que foram alimentados com α-tocoferol combinado com extrato de uva, mesmo quando não houve diferença no tamanho da lesão. CONCLUSÃO: Foi observado um efeito sinergístico entre os polifenóis e α-tocoferol, resultando na redução na evolução da aterosclerose e um maior de efeito benéfico na aterosclerose do que o consumo isolado de antioxidantes sobre a aterosclerose do que o consumo isolado de antioxidantes.

Apolipoprotein E polymorphism in cerebrovascular & coronary heart diseases.

The role of apolipoprotein E (apo E) in lipid metabolism and cholesterol transport is well established. About 14 per cent of the variation in plasma cholesterol levels is attributed to polymorphisms in apo E gene (APOapo E). E consists of three common alleles, designated as ε2, ε3 and ε4 which code for E2, E3 and E4 proteins respectively resulting in three homozygous (E2/E2, E3/E3, E4/E4) and three heterozygous (E3/E2, E4/E2 and E4/E3) phenotypes. Different populations studied worldwide inherit variable frequencies of the E alleles and genotypes, with the most frequent allele being ε3.The ε4 allele has been consistently shown to be associated with Alzheimer’s disease, coronary heart disease and cerebrovascular disorders. In this review, we have discussed the role of apo E polymorphisms in cerebrovascular and coronary heart diseases. The status of apo E polymorphisms and their disease associations in Asian Indians besides, other populations has also been discussed. Further, studies elucidating the pathophysiology of apo E deficiency conducted in knock-out mice have been reviewed.

ApoE polymorphisms and diarrheal outcomes in Brazilian shanty town children

A series of studies have shown that the heavy burdens of diarrheal diseases in the first 2 formative years of life in children living in urban shanty towns have negative effects on physical and cognitive development lasting into later childhood. We have shown that APOE4 is relatively common in shanty town children living in Brazil (13.4 percent) and suggest that APOE4 has a protective role in cognitive development as well as weight-for-height in children with heavy burdens of diarrhea in early childhood (64/123; 52 percent), despite being a marker for cognitive decline with Alzheimer’s and cardiovascular diseases later in life. APOE2 frequency was higher among children with heaviest diarrhea burdens during the first 2 years of life, as detected by PCR using the restriction fragment length polymorphism method, raising the possibility that ApoE-cholesterol balance might be critical for growth and cognitive development under the stress of heavy diarrhea burdens and when an enriched fat diet is insufficient. These findings provide a potential explanation for the survival advantage in evolution of genes, which might raise cholesterol levels during heavy stress of diarrhea burdens and malnutrition early in life.

Variantes de los genes CYP2C9 y apolipoproteína E en la respuesta individual a la warfarina / Variatins of gens CYO2C9 and apolipoprotein E in the induvidual response to warfarin

Introducción: la warfarina es el anticoagulante oral más usado en nuestro país y uno de los más usados a nivel mundial. Variantes polimórficas en varios genes influencian la respuesta individual a este fármaco, especialmente de CYP2C9 y de la apolipoproteína E. Nos proponemos estudiar la influencia en una población de pacientes uruguayos de estas variantes en la variabilidad interindividual en la respuesta a la warfarina y el riesgo de efectos adversos durante el tratamiento anticoagulante oral. Material y método: se incluyeron 55 pacientes en tratamiento crónico con warfarina. Se relevaron datos sobre dosis de mantenimiento, respuesta farmacocinética y efectos adversos. Se determinó el genotipo de los pacientes para las variantes *1, *2 y *3 de CYP2C9 y E2, E3 y E4 de apolipoproteína E. Resultados: los portadores de la variante *3 son los que requirieron dosis diarias de mantenimiento menores, seguidos por los portadores del alelo *2 y luego los homocigotos *1 (p=0,049). Los portadores de la variante *3 tuvieron también una mayor frecuencia de la tasa internacional normalizada (INR) por encima del rango deseado y requirieron más ajustes de dosis para lograr una adecuada anticoagulación; los eventos adversos fueron más frecuentes en los pacientes de genotipo *1/*3, tanto en relación al riesgo de sangrado como de sobreanticoagulación. La presencia del alelo E4 de la ApoE se asocia levemente a una mayor sensibilidad a la warfarina.Conclusiones: en el presente estudio, realizado en la primera población nacional proveniente de una policlínica de hipocoagulación, se confirma una mayor sensibilidad a la warfarina de los portadores de alelos variantes *2 y *3 de CYP2C9 y de la variante E4 de la ApoE, lo que puede tener importancia en la individualización de la dosis y los riesgos durante el tratamiento con este fármaco...

Introduction: warfarin is the most frequently used oral anticoagulant in Uruguay and worldwide. Polymorphic variants in various genes modulate individual response to this drug, especially CYP2C9. We studied the influence of these genetic variants on interindividual variability in response to warfarin and the risk of adverse reactions in an uruguayan population.Materials and methods: the study involved 55 patients undergoing chronic oral anticoagulant treatment with warfarin. Data on daily maintenance dose, pharmacokinetic response and adverse reactions was collected. CYP2C9 *1, *2 and *3, and Apoe E2, E3 and E4, genotype was determined by standard procedures.Results: carriers of CYP2C9 *3 allele required the lesser manteinance dose, followed by *2 allele carriers and then *1 homocygotes (p=0.049). CYP2C9 *3 allele carriers had more episodes of above de range International Normalized Ratio and required more dose adjustments to achieve a proper anticoagulation; adverse events were more frequent in patients with CYP2C9 *1/*3, regarding bleeding events as wells as overanticoagulation. Presence of ApoE E4 allele is associated with a moderately elevated sensibility to warfarin. Conclusions: this study is the first carried out on this subjet in an uruguayan population and confirms an increased sensibility to warfarin and risk of adverse effects in carriers of CYP2C9 *3 allele and in carriers of ApoE E4 variant, wich could be useful in warfarin dose and risk individualization during treatment with this drug. Genotypic data should be considered when establishing the best individual dose for individuals at high risk of bleeding, in those who suffered an adverse effect with this drug for physiopathlogic diagnosis and in young patients who underwent cardiac surgery who would potentially face a long term anticoagulant therapy. We will continue evaluating genomic components of risk during anticoagulation.

Effect of Lactobacillus delbrueckii on cholesterol metabolism in germ-free mice and on atherogenesis in apolipoprotein E knock-out mice

Elevated blood cholesterol is an important risk factor associated with atherosclerosis and coronary heart disease. Several studies have reported a decrease in serum cholesterol during the consumption of large doses of fermented dairy products or lactobacillus strains. The proposed mechanism for this effect is the removal or assimilation of intestinal cholesterol by the bacteria, reducing cholesterol absorption. Although this effect was demonstrated in vitro, its relevance in vivo is still controversial. Furthermore, few studies have investigated the role of lactobacilli in atherogenesis. The aim of the present study was to determine the effect of Lactobacillus delbrueckii on cholesterol metabolism in germ-free mice and the possible hypocholesterolemic and antiatherogenic action of these bacteria using atherosclerosis-prone apolipoprotein E (apo E) knock-out (KO) mice. For this purpose, Swiss/NIH germ-free mice were monoassociated with L. delbrueckii and fed a hypercholesterolemic diet for four weeks. In addition, apo E KO mice were fed a normal chow diet and treated with L. delbrueckii for 6 weeks. There was a reduction in cholesterol excretion in germ-free mice, which was not associated with changes in blood or liver cholesterol concentration. In apo E KO mice, no effect of L. delbrueckii was detected in blood, liver or fecal cholesterol. The atherosclerotic lesion in the aorta was also similar in mice receiving or not these bacteria. In conclusion, these results suggest that, although L. delbrueckii treatment was able to reduce cholesterol excretion in germ-free mice, no hypocholesterolemic or antiatherogenic effect was observed in apo E KO mice.

The beta-chemokines MIP-1alpha and RANTES and lipoprotein metabolism in HIV-infected brazilian patients

HIV patients are predisposed to the development of hypertriglyceridemia and hypercholesterolemia as a result of both viral infection and HIV infection therapy, especially the protease inhibitors. Chemokines and cytokines are present at sites of inflammation and can influence the nature of the inflammatory response in atherosclerosis. We investigated the correlation between biochemical variables and beta-chemokines (MIP-1alpha and RANTES) and the apolipoprotein E genotype in HIV-infected individuals. The apolipoproteins were measured by nephelometry. Triglycerides and total cholesterol were determined by standard enzymatic procedures. The beta-chemokines were detected by ELISA. The genetic category of CCR5 and apolipoprotein E were determined by PCR amplification and restriction enzymes. Immunological and virological profiles were assessed by TCD4+ and TCD8+ lymphocyte counts and viral load quantification. Positive correlations were found between apo E and CD8+ (p = 0.035), apo E and viral load (p = 0.018), MIP-1alpha and triglycerides (p = 0.039) and MIP-1a and VLDL (p = 0.040). Negative correlations were found between viral load and CD4+ (p = 0.05) and RANTES and CD4+ (p = 0.029). The beta-chemokine levels may influence lipid metabolism in HIV-infected individuals.

La apolipoproteina E: el polimorfismo genetico y su relacion con los cambios metabolicos, los habitos alimenticios y el origen etnico / Apolipoprotein E: its relationship with differents forms of genetic expression and effect of etnic origin and nutritional habits

La apolipoproteína E (Apo E) es una proteína plasmática constituyente de las lipoproteínas, que tiene como función mantener la estructura y regular el metabolismo de varias de ellas. Los tres alelos más comunes del gen de Apo E son e2, e3 y e4, los cuales producen tres isoformas de la proteína, llamadas E2, E3 y E4. Estos tres alelos diferentes son heredados en forma codominante dando como resultado seis genotipos: E2/2, E3/2, E3/3, E3/4, E4/4, E4/E2. El genotipo E3/E3 es el normal, con una frecuencia poblacional de 77 por ciento. Se ha estimado que 60 por ciento de la variación plasmática del colesterol está determinada genéticamente; el polimorfismo de Apo E corresponde a 14 por ciento de esas variaciones genéticas. La deficiencia de Apo E causa hipercolesterolemia severa. Genotipos específicos han sido implicados en el desarrollo de aterosclerosis, de enfermedad coronaria, de hiperlipoproteinemia tipo III (HLP tipo III), de enfermedad cerebrovascular y de formas familiares y esporádicas de enfermedad de Alzheimer. Esta revisión muestra diferentes aspectos bioquímicos y genéticos de la apolipoproteína E, así como la relación de las diferentes formas de expresión genética de esta proteína con los cambios metabólicos de las lipoproteínas y con el origen étnico y los hábitos alimenticios

Polimorfismo da apolipoproteina E na doenca de Alzheimer tipo tardio: resultados preliminares / Apolipoproteins E polymorphism in Alzheimer's disease: delayed type: preliminary results

A apolipoproteina E (apo E), com funcao no metabolismo de lipideos e transporte de colesterol, tem sido associada tambem a patogenese da doenca de Alzheimer (DA). Este estudo teve como objetivo identificar os genotipos para apo E e a frequencia de seus alelos em individuos com DA tipo tardio (grupo 1) ou sem sintomas de demencia neurodegenerativa (grupo 2). Foram estudados 32 individuos assim distribuidos: grupo 1 = 18 pacientes com idades de 66 a 82 anos (media = 71 anos); grupo 2 = 14 individuos de 65 a 78 anos (media = 69 anos). O DNA foi extraido de leucocitos com amplificacao do segmento de interesse do gene para apo E por PCR (polymerase chain reaction) e submetido a clivagem com enzima Hha I. As frequencias dos alelos nos grupos 1 e 2 foram: 3 - 0,75 e 0,79 (p=0,699), 4 - 0,25 e 0,07 (p=0,035), respectivamente...