Prevention of the development of heart failure with preserved ejection fraction by the phosphodiesterase-5A inhibitor vardenafil in rats with type 2 diabetes

AIMS: Heart failure with preserved ejection fraction (HFpEF) has a great epidemiological burden. The pathophysiological role of cyclic guanosine monophosphate (cGMP) signalling has been intensively investigated in HFpEF. Elevated levels of cGMP have been shown to exert cardioprotective effects in various cardiovascular diseases, including diabetic cardiomyopathy. We investigated the effect of long-term preventive application of the phosphodiesterase-5A (PDE5A) inhibitor vardenafil in diabetic cardiomyopathy-associated HFpEF. METHODS AND RESULTS: Zucker diabetic fatty (ZDF) rats were used as a model of HFpEF and ZDF lean rats served as controls. Animals received vehicle or 10 mg/kg body weight vardenafil per os from weeks 7 to 32 of age. Cardiac function, morphology was assessed by left ventricular (LV) pressure-volume analysis and echocardiography at week 32. Cardiomyocyte force measurements were performed. The key markers of cGMP signalling, nitro-oxidative stress, apoptosis, myocardial hypertrophy and fibrosis were examined. The ZDF animals showed diastolic dysfunction (increased LV/cardiomyocyte stiffness, prolonged LV relaxation time), preserved systolic performance, decreased myocardial cGMP level coupled with impaired protein kinase G (PKG) activity, increased nitro-oxidative stress, enhanced cardiomyocyte apoptosis, and hypertrophic and fibrotic remodelling of the myocardium. Vardenafil effectively prevented the development of HFpEF by maintaining diastolic function (decreased LV/cardiomyocyte stiffness and LV relaxation time), by restoring cGMP levels and PKG activation, by lowering apoptosis and by alleviating nitro-oxidative stress, myocardial hypertrophy and fibrotic remodelling. CONCLUSIONS: We report that vardenafil successfully prevented the development of diabetes mellitus-associated HFpEF. Thus, PDE5A inhibition as a preventive approach might be a promising option in the management of HFpEF patients with diabetes mellitus

Morphometric Study of Diabetes Related Alterations in Human Parotid Gland and Comparison with Submandibular Gland

Type 2 diabetes mellitus represents one of the principal diseases that afflict the world population and is often associated with malfunction of salivary glands and consequent oral diseases. We recently described significant ultrastructural alterations in the human submandibular gland in diabetic patients without evident oral pathologies. Herein, an analogs morphometrical investigation was focused on the parotid gland in order to evaluate if one of the two glands is more affected by diabetes. Parotid fragments from diabetic and nondiabetic patients were fixed, dehydrated, and processed for light and electron microscopy. Serous cells were randomly photographed and the density and size of several structures involved in the secretory process were examined by morphometry. Scanning electron microscopy images revealed significant changes in the number of apically docked granules and vesicles, suggesting that the last steps in exocytosis are somehow altered in diabetic cells. Other variables analyzed by light and transmission electron microscopy such as the size of acini and secretory granules did not show significant changes, but comparison with previous data obtained with submandibular gland cells demonstrated that the two glands are affected differently. Anat Rec, 2015. © 2015 Wiley Periodicals, Inc

Sarco(endo)plasmic Reticulum Ca2+-ATPase-2 Gene: Structure and Transcriptional Regulation of the Human Gene

The sarco(endo)plasmic reticulum Ca2+-ATPases (SERCAs) belong to a family of active calcium transport enzymes encoded by the SERCA1, 2, and 3 genes. In this study, we describe the complete structure of the human SERCA2 gene and its 5’ -regulatory region. The hSERCA2 gene is located in chromosome 12 position q24.1 in Contig NT_009770.8, spans 70 kb, and is organized in 21 exons intervened by 20 introns. The last two exons of the pre-mRNA produce by alternatively splicing the cardiac/slow-twitch muscle-specific SERCA2a isoform and the ubiquitous SERCA2b isoform. The sequence of the proximal 225-bp regulatory region of the SERCA2 genes is 80% G+C-rich and is conserved among human, rabbit, rat, and mouse species. It contains a TATA-like-box, an E-box/USF sequence, a CAAT-box, four Sp1 binding sites, and a thyroid hormone responsive element (TRE). There are two other conserved regulatory regions located between positions -410 to -661 bp and from -919 to -1410 bp. Among the DNA cis-elements present in these two regulatory regions there are potential binding sites for: GATA-4, -5, -6, Nkx-2.5/Csx, OTF-1, USF, MEF-2, SRF, PPAR/RXR, AP-2, and TREs. Upstream from position -1.5 kb, there is no significant homology among the SERCA2 genes cloned. In addition, the human gene has several repeated sequences mainly of the Alu and L2 type located upstream from position -1.7 kb, spanning in a continuous fashion for more than 40 kb. In this study, we report the cloning of 2.4 kb of 5’-regulatory region and demonstrate that the proximal promoter region is sufficient for expression in cardiac myocytes, and the region from -225 to -1232 bp contains regulatory DNA elements which down-regulate the expression of the SERCA2 gene in neonatal cardiomyocytes

Función del retículo sarcoplásmico y su papel en las enfermedades cardíacas

El retículo sarcoplásmico (RS) es el principal almacén de calcio intracelular en el músculo estriado y participa de forma importante en la regulación del proceso acoplamiento-excitación-contracción (AEC) en el músculo esquelético y cardíaco, regulando las concentraciones intracelulares de calcio durante la contracción y la relajación muscular. Esta regulación está dada por la interacción de las principales proteínas del RS que son el canal de liberación de calcio o receptor de rianodina, la ATPasa de Ca2+, fosfolamban y calsecuestrina. Por la relevancia del AEC en la fisiopatología de varias enfermedades cardíacas, se ha estudiado extensamente el papel que mantiene el RS y sus distintos componentes proteicos en distintas patologías, principalmente en la hipertrofia cardíaca, la insuficiencia cardíaca y en las arritmias hereditarias. Por lo anterior, las proteínas del RS constituyen un área de gran interés para el desarrollo de nuevas terapias, por lo que resulta de gran importancia el comprender la función del RS. En este artículo de revisión se analiza la estructura y función de las principales proteínas del RS, su papel en los procesos de contracción y relajación muscular, así como los cambios en expresión y función que ocurren en diferentes patologías cardíacas

Terapia génica en la insuficiencia cardiaca

La terapia génica como estrategia de tratamiento para la insuficiencia cardiaca es un área en la que en los últimos 10 años la investigación se ha incrementado importantemente; es una de las de mayor promesa para obtener una terapia exitosa para la insuficiencia cardiaca, pues ofrece la posibilidad de corregir los defectos básicos de la enfermedad a escala celular. Uno de los primeros pasos a considerar en el uso de esta terapia es la forma de administrar el material genético; los vectores que han mostrado mayor utilidad en el miocardio son los adenovirus y los virus adeno-asociados; la administración local ha demostrado ser superior a la administración periférica. Se han identificado distintos mecanismos fisiopatológicos, susceptibles de ser modificados por terapia génica, entre los que se encuentran la regulación de los flujos de Ca2+ durante el proceso del acoplamiento excitación-contracción, alteraciones en la señalización intracelular y en el sistema adrenérgico, el desarrollo de apoptosis, y la angiogénesis. En la actualidad ya existen datos de ensayos clínicos en humanos empleando algunas de estas estrategias. El objeto de la presente revisión es hacer un recuento de los datos existentes en la literatura, así como de las perspectivas a futuro