10 research outputs found
Papel de la mitocondria en la cardiopatía isquémica : estudio de asociación con haplogrupos mitocondriales y análisis del proteoma mitocondrial de los corazones patológicos
[Resumen] ANTECEDENTES. Los eventos que suceden en la mitocondria juegan un papel fundamental en la cardiopatía isquémica (CI).
OBJETIVOS. El primer objetivo de este estudio ha sido analizar la influencia de los principales haplogrupos europeos y los polimorfismos que los definen sobre el desarrollo de la CI en una población española. El segundo objetivo ha sido investigar el perfil proteico mitocondrial de la región ventricular sana, próxima a la zona de infarto en un modelo porcino de isquemia y reperfusión (I/R).
MÉTODOS. Para el primer objetivo, se detectaron los polimorfismos que definen los distintos haplogrupos mitocondriales en muestras de ADN de 423 controles y 358 pacientes con CI. Para el segundo se extrajeron proteínas mitocondriales de tejido cardíaco de 11 cerdos (Sus scrofa) 3 o 15 días tras el procedimiento de I/R. Los cambios en la expresión de las proteínas mitocondriales con respecto al grupo control de cerdos sin operar fueron detectados mediante espectrometría de masas en tándem.
RESULTADOS. En el primer estudio, el haplogrupo H se encontró significativamente sobrerrepresentado en el grupo patológico (OR=1,50, p<0,05) al mismo tiempo que la frecuencia del haplogrupo J en el grupo de pacientes con CI fue significativamente menor que en la población control (OR=0,47, p<0,05). En el análisis del proteoma mitocondrial, se identificaron 70 proteínas mitocondriales significativamente reguladas. Se observó un incremento de la expresión de proteínas implicadas en la cadena de transporte de electrones y en la glucólisis a los 3 dias, como posible mecanismo para compensar la demanda energética en las zonas próximas a la lesión. A los 15 días el patrón proteico revirtió hacia uno más próximo al observado en animales control, sugiriendo un sesgo hacia la estabilización del tejido. Se validó por primera vez la sobreexpresión de la proteína BRP44L, un transportador de piruvato, que podría ser esencial para la obtención de energía tras la isquemia.
CONCLUSIONES. 1) Se ha demostrado por primera vez que haplogrupos con fenotipos opuestos en relación con la producción de especies reactivas de oxígeno son respectivamente factores de riesgo (H) y de protección (J) para el desarrollo de CI. 2) Se han determinado diferencias en el perfil proteómico mitocondrial 3 y 15 días tras la I/R mostrando un aumento de proteínas de transporte y metabolismo de piruvato e involucradas en la producción de enenrgía. El estudio en profundidad de proteínas detectadas en este estudio, como BRP44L, podrían ser útiles para el desarrollo de terapias enfocadas a la estabilización del tejido miocárdico en etapas post-isquémicas.[Resumo] FONDO. Os eventos que suceden na mitocondria xogan un papel fundamental na cardiopatía isquémica (CI).
OBXECTIVOS. O primeiro obxectivo deste estudo foi analizar a influencia dos principais haplogrupos europeos e os polimorfismos que os definen sobre o desenvolvemento da CI nunha poboación española. O segundo obxectivo foi o de investigar o perfil proteico mitocondrial da rexión ventricular sá, próxima á zona de infarto nun modelo porcino de isquemia e reperfusíon (I/R).
MÉTODOS. Para o primeiro obxectivo, detectáronse os polimorfismos que definen os distintos haplogrupos mitocondriais en mostras de ADN de 423 controles e 358 doentes con CI. Para o segundo extraéronse proteínas mitocondriais de texido cardíaco de 11 cerdos (Sus scorfa) 3 ou 15 días tras o procedemento de I/R. Os cambios na expresión das proteínas mitocondriais con respecto ó grupo control de cerdos sen operar foron detectados mediante espectometría de masas en tándem.
RESULTADOS. No primeiro estudo, o haplogrupo H atopouse significativamente máis representado no grupo patolóxico (OR=1,50, p<0,05) ao mesmo tempo que a frecuencia do haplogrupo J no grupo de doentes con CI foi significativamente menor que na poboación control (OR=0,47, p<0,05). No análise do proteoma mitocondrial identificarónse 70 proteínas mitocondriais significativamente reguladas. Observouse un incremento da expresión de proteínas implicadas na cadena de transporte de eletróns e na glucólise ós 3 días como posible mecanismo para compensar a demanda enerxética das zonas próximas á lesión. Ós 15 días o patrón proteico revertiu a un máis próximo ó observado en animais control, suxerindo un sesgo cara a estabilización do texido. Validóuse por primeira vez a sobreexpresión da proteina BRP44L, un transportador de piruvato que poderia ser esencial para a obtención de enerxia tras a isquemia.
CONCLUSIÓNS. 1) Demostrouse por primeira vez que haplogrupos con fenótipos opostos en relación á produción de especies reactivas de osíxeno son, respectivamente, factores de risco (H) e de protección (J) para o desenvolvemento da CI. 2) Determináronse diferencias no perfil proteico mitocondrial 3 e 15 días tras a I/R amosando un aumento de proteínas de transporte e metabolismo de piruvato e involucradas na producción de enerxía. O estudo en profundidade das proteínas detectadas neste estudo, coma BRP44L, poderían ser utilies para o desenvolvemento de terapias enfocadas á estabilización do texido miocárdico en etapas post-isquémicas.[Abstract] BACKGROUND. The events that occur in the mitochondria play a key role in ischemic cardiomyopathy (IC).
OBJECTIVES. The first objective of the study was to analyze the influence of major European haplogroups and polymorphisms that define the development of IC in a Spanish population. The second objective was to investigate the mitochondrial protein profile of a healthy ventricular region, adjacent to the site of infarction in a porcine model of ischemia and reperfusion (I/R).
METHODS. To accomplish the first aim the polymorphisms that define the different mitochondrial haplogroups were detected in DNA samples of 423 controls and 358 patients with IC. For the second objective mitochondrial proteins were extracted from heart tissue from 11 pigs (Sus scrofa), 3 or 15 days after I/R. Changes in mitochondrial protein expression were detected and compared against non-operated pigs by tandem mass spectrometry.
RESULTS. In the first study, haplogroup H was significantly overrepresented in the pathological group (OR=1.50, p<0.05) while the frequency of haplogroup J in the group of patients with IC was significantly lower compared against the control population (OR=0.47, p<0.05). In the analysis of the mitochondrial proteome, we identified 70 significantly regulated mitochondrial proteins. An increase in the expression of proteins involved in the electron transport chain and glycolysis were observed 3 days post I/R. This is a possible mechanism to compensate for the energy demand in the areas surrounding the lesion. Interestingly, the protein expression pattern had changed dramatically after 15 days, reverting to a similar pattern to that observed in control animals, suggesting a bias toward tissue stabilization. The overexpression of BRP44L, a pyruvate transporter that could be essential for the energy uptake after the ischemia, was validated for the first time.
CONCLUSIONS. 1) This study has demonstrated for the first time that haplogroups with opposite phenotypes regarding the production of reactive oxygen species are respectively risk (H) and protective factors (J) for the development of IC. 2) Differences in mitochondrial proteomic profile 3 and 15 days post I/R have been determined showing an increase exprerssion of proteins involved with pyruvate transport, metabolism and energy production. An in-depth analysis of proteins detected in this study such as BRP44L, could be useful for the development of therapies aimed at stabilizing the myocardial tissue in post-ischemic stages
Mitochondrial DNA haplogroup H as a risk factor for idiopathic dilated cardiomyopathy in Spanish population
[Abstract] Idiopathic dilated cardiomyopathy (IDC) is a structural heart disease with strong genetic background. The different single nucleotide polymorphisms (SNPs) that constitute mitochondrial haplogroups could play an important role in IDC progression. The aim of this study was to test frequencies of mitochondrial haplogroups in healthy controls (n = 422) and IDC patients (n = 304) of a Caucasian Spanish population. To achieve this, ten major European haplogroups were identified. Frequencies and Odds Ratios for the association between IDC and haplogroups were calculated in both groups. We found that compared to healthy controls, the prevalence of haplogroup H was significantly higher in IDC patients (40.0% vs 50.7%, p-value = 0.040).Instituto de Salud Carlos III; PS09/0084
Mitochondrial haplogroups H and J: risk and protective factors for ischemic cardiomyopathy
[Abstract] Background. Since mitochondria are the principal source of reactive oxygen species (ROS), these organelles may play an important role in ischemic cardiomyopathy (IC) development. The mitochondrial genome may influence this disease. The aim of the present study was to test the relationship between IC development and the impact of single nucleotide polymorphisms (SNPs) in mitochondrial DNA (mtDNA) defining the mitochondrial haplogroups in a population study.
Methodology and principal findings. Ten major European haplogroups were identified by using the single base extension technique and by polymerase chain reaction-restriction fragment length polymorphism. Frequencies and Odds Ratios for the association between IC patients (n = 358) and healthy controls (n = 423) were calculated. No convincing associations between classical risk factors for ischemic cardiomyopathy development and haplogroups were found. However, compared to healthy controls, the prevalence of haplogroup H was significantly higher in IC patients (40.0% vs 50.0%, p-value = 0.039) while the frequency of haplogroup J was significantly lower (11.1% vs 5.6%, p-value = 0.048). The analysis of the SNPs characterizing the European mtDNA haplogroups showed that the m.7028C allele (40.0% vs 50.0%, p-value = 0.005) and m.14766C allele (43.0% vs 54.2%, p-value = 0.002) were overrepresented in IC patients, meanwhile the m.10398G allele (19.8% vs 13.1%, p-value = 0.015) and m.4216C allele (22.2% vs 16.5%, p-value = 0.044) were found as protective factors against IC.
Conclusions and significance. Our results showed that the haplogroups H and J were found as a risk and protective factors for ischemic cardiomyopathy development, respectively.Instituto de Salud Carlos III; PS09/0084
Dynamic and complex redox-dependent modifications of DJ-1 in cardiac cells and tissue during oxidative stress
Proteomics analysis of cardiac extracellular matrix remodeling in a porcine model of ischemia/reperfusion injury
Analysis of mitochondrial proteins in the surviving myocardium after ischemia identifies mitochondrial pyruvate carrier expression as possible mediator of tissue viability
Instituto de Salud Carlos III; PS09/0084
Mitochondrial Haplogroups H and J: Risk and Protective Factors for Ischemic Cardiomyopathy
Primer sequences used for in multiplex PCR, SBE and PCR-RFLP.
<p>R: primer in reverse orientation; F:primer in forward orientation.</p>*<p>Lower case letters indicate the unspecific nucleotides in 5′-end of the SBE primer. PCR products for RFLP analysis were digested with the corresponding restriction enzyme and digested PCR products appeared like three fragments in agarose gel.</p
Multivariate analysis of the study groups.
<p>B. Regression coefficient. SEM. Standart error of the mean. OR. Odds Ratio. 95%CI. Confidence Intervals.</p><p>Significant differences (p-value<0.05) are indicated in bold.</p