Novel mechanism of hydrogen sulfide-induced guinea pig urinary bladder smooth muscle contraction: role of BK channels and cholinergic neurotransmission

Hydrogen sulfide (H2S) is a key signaling molecule regulating important physiological processes, including smooth muscle function. However, the mechanisms underlying H2S-induced detrusor smooth muscle (DSM) contractions are not well understood. This study investigates the cellular and tissue mechanisms by which H2S regulates DSM contractility, excitatory neurotransmission, and large-conductance voltage- and Ca2+-activated K+ (BK) channels in freshly isolated guinea pig DSM. We used a multidisciplinary experimental approach including isometric DSM tension recordings, colorimetric ACh measurement, Ca2+ imaging, and patch-clamp electrophysiology. In isolated DSM strips, the novel slow release H2S donor, P-(4-methoxyphenyl)-p-4-morpholinylphosphinodithioic acid morpholine salt (GYY4137), significantly increased the spontaneous phasic and nerve-evoked DSM contractions. The blockade of neuronal voltage-gated Na+ channels or muscarinic ACh receptors with tetrodotoxin or atropine, respectively, reduced the stimulatory effect of GYY4137 on DSM contractility. GYY4137 increased ACh release from bladder nerves, which was inhibited upon blockade of L-type voltage-gated Ca2+ channels with nifedipine. Furthermore, GYY4137 increased the amplitude of the Ca2+ transients and basal Ca2+ levels in isolated DSM strips. GYY4137 reduced the DSM relaxation induced by the BK channel opener, NS11021. In freshly isolated DSM cells, GYY4137 decreased the amplitude and frequency of transient BK currents recorded in a perforated whole cell configuration and reduced the single BK channel open probability measured in excised inside-out patches. GYY4137 inhibited spontaneous transient hyperpolarizations and depolarized the DSM cell membrane potential. Our results reveal the novel findings that H2S increases spontaneous phasic and nerve-evoked DSM contractions by activating ACh release from bladder nerves in combination with a direct inhibition of DSM BK channels.National Institute of Diabetes and Digestive and Kidney DiseasesDepto. de FisiologíaFac. de FarmaciaTRUEpu

Cardiac System during the Aging Process

The aging process is accompanied by a continuous decline of the cardiac system, disrupting the homeostatic regulation of cells, organs, and systems. Aging increases the prevalence of cardiovascular diseases, thus heart failure and mortality. Understanding the cardiac aging process is of pivotal importance once it allows us to design strategies to prevent age-related cardiac events and increasing the quality of live in the elderly. In this review we provide an overview of the cardiac aging process focus on the following topics: cardiac structural and functional modifications; cellular mechanisms of cardiac dysfunction in the aging; genetics and epigenetics in the development of cardiac diseases; and aging heart and response to the exercise.Sección Deptal. de Fisiología (Farmacia)Depto. de EnfermeríaFac. de FarmaciaFac. de Enfermería, Fisioterapia y PodologíaTRUEpu

Role of Calcitonin Gene-Related Peptide in Inhibitory Neurotransmission to the Pig Bladder Neck

Purpose: We studied the role of calcitonin gene-related peptide in nonadrenergic, noncholinergic neurotransmission to the pig bladder neck. Materials and Methods: We used immunohistochemical techniques to determine the distribution of calcitonin gene-related peptide immunoreactive fibers as well as organ baths for isometric force recording. We investigated relaxation due to endogenously released or exogenously applied calcitonin gene-related peptide in urothelium denuded phenylephrine precontracted strips treated with guanethidine, atropine and NG-nitro-L-arginine to block noradrenergic neurotransmission, muscarinic receptors and nitric oxide synthase, respectively. Results: Rich calcitonin gene-related peptide immunoreactive innervation was found penetrating through the adventitia and distributed in the suburothelial and muscle layers. Numerous, variable size, varicose calcitonin gene-related peptide immunopositive terminals were seen close below the urothelium. In the muscle layer calcitonin gene-related peptide immunopositive nerves usually appeared as varicose terminals running along muscle fibers. Electrical field stimulation (2 to 16 Hz) and exogenous calcitonin gene-related peptide (0.1 nM to 0.3 μM) evoked frequency and concentration dependent relaxation, respectively. Nerve responses were potentiated by capsaicin, decreased by calcitonin gene-related peptide (8–37) and abolished by tetrodotoxin, capsaicin sensitive primary afferent blockers, calcitonin gene-related peptide receptors and neuronal voltage gated Na+ channels. Calcitonin gene-related peptide-induced relaxation was potentiated by the neuronal voltage gated Ca2+ channels blocker ω-conotoxin-GVIA and decreased by calcitonin gene-related peptide (8–37). Calcitonin gene-related peptide relaxation was not modified by blockade of endopeptidases, nitric oxide synthase, guanylyl cyclase and cyclooxygenase. Conclusions: Results suggest that calcitonin gene-related peptide is involved in the nonadrenergic, noncholinergic inhibitory neurotransmission of the pig bladder neck, producing relaxation through neuronal and muscle calcitonin gene-related peptide receptors. Nitric oxide/cyclic guanosine monophosphate and cyclooxygenase pathways do not seem to be involved in such responses.Ministerio de Ciencia e Innovación, SpainSección Deptal. de Fisiología (Farmacia)Fac. de FarmaciaTRUEpu

Hydrogen sulfide mediated inhibitory neurotransmission to the pig bladder neck: role of KATP channels, sensory nerves and calcium signaling.

Purpose: Because neuronal released endogenous H2S has a key role in relaxation of the bladder outflow region, we investigated the mechanisms involved in H2S dependent inhibitory neurotransmission to the pig bladder neck. Materials and methods: Bladder neck strips were mounted in myographs for isometric force recording and simultaneous measurement of intracellular Ca(2+) and tension. Results: On phenylephrine contracted preparations electrical field stimulation and the H2S donor GYY4137 evoked frequency and concentration dependent relaxation, which was reduced by desensitizing capsaicin sensitive primary afferents with capsaicin, and the blockade of adenosine 5'-triphosphate dependent K(+) channels, cyclooxygenase and cyclooxygenase-1 with glibenclamide, indomethacin and SC560, respectively. Inhibition of vanilloid, transient receptor potential A1, transient receptor potential vanilloid 1, vasoactive intestinal peptide/pituitary adenylyl cyclase-activating polypeptide and calcitonin gene-related peptide receptors with capsazepine, HC030031, AMG9810, PACAP6-38 and CGRP8-37, respectively, also decreased electrical field stimulation and GYY4137 responses. H2S relaxation was not changed by guanylyl cyclase, protein kinase A, or Ca(2+) activated or voltage gated K(+) channel inhibitors. GYY4137 inhibited the contractions induced by phenylephrine and by K(+) enriched (80 mM) physiological saline solution. To a lesser extent it decreased the phenylephrine and K(+) induced increases in intracellular Ca(2+). Conclusions: H2S produces pig bladder neck relaxation via activation of adenosine 5'-triphosphate dependent K(+) channel and by smooth muscle intracellular Ca(2+) desensitization dependent mechanisms. H2S also promotes the release of sensory neuropeptides and cyclooxygenase-1 pathway derived prostanoids from capsaicin sensitive primary afferents via transient receptor potential A1, transient receptor potential vanilloid 1 and/or related ion channel activation. Keywords: 4-AP; 4-aminopyridine; AM; ATP; ATP dependent K(+); CGRP; COX; CSE; CSPA; Ca(2+) activated K(+); Emax; K(ATP); K(Ca); K(V); KPSS; L-NOARG; MLCP; N(G)-nitro-L-arginine; NO; PACAP; PKA; PSS; TRPA(1); TRPV(1); VOC; VPAC; [Ca(2+)](i); acetoxymethyl ester; adenosine 5′-triphosphate; calcitonin gene-related peptide; capsaicin sensitive primary afferent; cyclooxygenase; cystathionine γ-lyase; hydrogen sulfide; intracellular Ca(2+); maximum response; muscle, smooth; myosin light chain phosphatase; nitric oxide; physiological saline solution; pituitary adenylyl cyclase activating polypeptide; potassium channels; potassium rich PSS; protein kinase A; synaptic transmission; transient receptor potential A1; transient receptor potential vanilloid 1; urinary bladder; vasoactive intestinal peptide receptor; voltage gated Ca(2+); voltage gated K(+).Depto. de EnfermeríaFac. de Enfermería, Fisioterapia y PodologíaTRUEunpu

Neuronal and non-neuronal bradykinin receptors are involved in the contraction and/or relaxation to the pig bladder neck smooth muscle

Aims: The current study investigates the role played by bradykinin (BK) receptors in the contractility to the pig bladder neck smooth muscle. Methods: Bladder neck strips were mounted in myographs for isometric force recordings and BK receptors expression was also determined by immunohistochemistry. Results: B2 receptor expression was observed in the muscular layer and urothelium whereas B1 expression was consistent detected in urothelium. A strong B2 immunoreactivity was also observed within nerve fibers among smooth muscle bundles. On urothelium-denuded preparations basal tone, BK induced concentration-dependent contractions which were reduced in urothelium-intact samples, by extracellular Ca(2+) removal and by blockade of B2 receptors and voltage-gated Ca(2+) (VOC) and non-VOC channels, and increased by cyclooxygenase (COX) inhibition. On phenylephrine-precontracted denuded strips, under non-adrenergic non-cholinergic (NANC) conditions, electrical field stimulation-elicited frequency-dependent relaxations which were reduced by B2 receptor blockade. In urothelium-intact samples, the B1 receptor agonist kallidin promoted concentration-dependent relaxations which were reduced by blockade of B1 receptors, COX, COX-1 and large-conductance Ca(2+) -activated K(+) (BKCa ) channels and abolished in urothelium-denuded samples and in K(+) -enriched physiological saline solution-precontracted strips. Conclusions: These results suggest that BK produces contraction of pig bladder neck via smooth muscle B2 receptors coupled to extracellular Ca(2+) entry via VOC and non-VOC channels with a minor role for intracellular Ca(2+) mobilization. Facilitatory neuronal B2 receptors modulating NANC inhibitory neurotransmission and urothelial B1 receptors producing relaxation via the COX-1 pathway and BKCa channel opening are also demonstrated.Ministerio de Ciencia e Innovación (España)Fundação para a Ciência e Tecnologia, Ministério da Educação e Ciência (Portugal)Depto. de FisiologíaFac. de FarmaciaTRUEpu

Self-assessment, Co-assessment, and the use of ICT as an innovative approach in Physiopathology practices and their effect on the student's teaching-learning process

En las últimas décadas, la educación universitaria ha evolucionado hacia un enfoque constructivista en consonancia con las recomendaciones del Espacio Europeo de Educación Superior (EEES). En este paradigma, los estudiantes asumen un papel activo en el proceso de enseñanza-aprendizaje, mientras los profesores actúan como facilitadores. Las metodologías constructivistas fomentan el desarrollo tanto individual como grupal de competencias específicas y genéricas, al tiempo que permiten la inclusión de agentes de evaluación formativa para estimular la crítica y la autocrítica del alumno en su desempeño. En este contexto, surge la necesidad de aplicar el constructivismo a la evaluación, involucrando al estudiante en su propio proceso de evaluación. La autoevaluación y la coevaluación emergen como alternativas concretas para lograrlo. La autoevaluación implica que el estudiante analice y valore de manera sistemática su trabajo durante el proceso de aprendizaje para mejorar resultados y fomentar la autocrítica. Por otro lado, la coevaluación es una evaluación entre compañeros que permite valorar la implicación y actitud de los miembros del grupo, estimulando el aprendizaje colectivo. Las Tecnologías de la Información y Comunicación (TIC) juegan un papel importante en la educación y en la evaluación de los alumnos, diferenciándose de las prácticas tradicionales. La implementación de TIC no solo desarrolla habilidades en el proceso enseñanza-aprendizaje, sino también favorece la autoevaluación y la coevaluación. Con base en este enfoque, se presenta un proyecto de innovación docente en la asignatura de Fisiopatología para estudiantes de Farmacia. Los alumnos crearán videos sobre temas específicos de la práctica y se evaluarán a sí mismos y a sus compañeros utilizando la herramienta App Plickers. Sin embargo, aún no existe una metodología claramente definida para la implementación de estrategias constructivistas y uso de TIC en Fisiopatología, destacando la importancia y relevancia de este proyecto.Depto. de FisiologíaFac. de FarmaciaFALSEsubmitte

Stop Hypertension with SLP/UCM I

El Proyecto Stop Hipertensión con ApS/UCM pretende aportar nuestro granito de arena a la lucha contra la hipertensión arterial (HTA), utilizando una herramienta educativa de aprendizaje en servicio (ApS), que permita a los estudiantes de los Grados en ciencias de la salud de la UCM aprender a la vez que realizan un servicio público. La hipertensión arterial (HTA) es un problema de salud pública y un importante factor de riesgo de padecer enfermedades cardiovasculares (ECV). Su diagnóstico está determinado por la medida de la presión arterial (PA) por lo que es de extrema importancia asegurar una medida fiable y válida. La automedida de la PA (AMPA) consiste en la medida de la PA por el propio paciente o un familiar, habitualmente en su domicilio, y es una herramienta útil para el diagnóstico y control de la HTA. Pero ¿sabemos medirnos de forma correcta la PA? ¿Usamos bien los tensiómetros? ¿Sabemos interpretar los registros de la PA y cuándo consultar al médico? Realizar AMPA correctamente no es fácil y exige un esfuerzo extraordinario de recursos humanos y económicos por parte de los profesionales de Atención Primaria. Por ello son necesarias nuevas estrategias para capacitar a la población en AMPA. Con este Proyecto nos proponemos luchar contra la HTA a través de la metodología educativa de aprendizaje-servicio que combina objetivos académicos con el servicio comunitario. El alumnado de los grados de Ciencias de la Salud de la UCM forman a la población en la correcta AMPA, asesorándoles sobre el uso correcto de los tensiómetros, enseñándoles a interpretar los resultados y alertándoles de cuándo acudir al médico. En el curso 2022-2023, el servicio público se ha dirigido principalmente al colectivo de mayores por ser especialmente vulnerables a padecer ECV en los Ayuntamientos de Coslada, Alcobendas y Alcorcón, y en las residencias Afanias y Neurovida, y a la población general que acuda a la Facultad durante la Semana de la Ciencia CAM/UCM 2022, a la Feria de la Salud de Coslada y a la campaña contra la HTA de la UCM junto con el Servicio de Medicina del Trabajo. Hemos demostrado que este proyecto es una herramienta rentable y efectiva en la lucha contra la HTA, vinculando la salud pública, los recursos de los ayuntamientos y la universidad. Se cubre así una necesidad y servicio social que podría salvar muchas vidas con recursos de bajo costo, cumpliendo con los objetivos del desarrollo sostenible y con los objetivos de la Sociedad Española de Hipertensión (SEH-LELHA) y de la estrategia HEARTS de la OPS/OMS. Los estudiantes se dan cuenta de la realidad de la Salud pública al tener contacto directo con la población, realizando un servicio a la sociedad, y desarrollándose profesionalmente. Agrademos su colaboración a: Los ayuntamientos de Coslada, Alcorcón y Alcobendas de Madrid. Las residencias de mayores Afanias y Neurovida. Las Sociedades científicas SEH-LELHA, SECF y SEAPEC. El Dr. Orduñez, lider de la estrategia HEARTS PAHO/WHO Los colegios profesionales COFM y CODEM. Las empresas River International S.L. (Beurer) and Peroxfarma S.L. (Omrom)Oficina APS UCMDepto. de FisiologíaFac. de FarmaciaTRUEunpu

Núcleos de Ensino da Unesp: artigos 2008

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq