Efecto del uso de resveratrol, quercetina y ácido gálico, antioxidantes encontrados en orujo de Vitis Vinífera, sobre la movilidad, función mitocondrial y capacidad fertilizadora de espermatozoides humanos
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Fecha
2012
Autores
Profesor/a Guía
Facultad/escuela
Idioma
es
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Universidad Andrés Bello
Nombre de Curso
Licencia CC
Licencia CC
Resumen
Actualmente, se ha observado una alarmante disminución de la calidad seminal en todo
el mundo afectando directamente la posibilidad de tener hijos. Esta disminución de la
calidad seminal es probablemente causada por el estilo de vida que llevamos, que
conlleva a la generación de radicales libres. Estas sustancias son altamente reactivas y
presentan una gran afinidad por membranas, convirtiéndolas en verdaderas
destructoras celulares. Para combatir estos radicales libres es natural pensar en la
utilización de antioxidantes. Estas son sustancias capaces de eliminar a los radicales
libres y de esta forma ayudar en la lucha contra enfermedades y malestares de la vida
cotidiana, reduciéndolos. La hipótesis planteada es que la utilización de resveratrol ,
quercetina y ácido gálico, antioxidantes provenientes del orujo de la vid ( Vitis vinífera) ,
mejora movilidad progresiva, función mitocondrial y capacidad fertilizadora de
espermatozoides provenientes de muestras de pacientes normales
(normozoospermicos) y sobre muestras anormales (astenozoospermicos). Para
comprobar esto se propone la aplicación de una solución mezcla de antioxidantes a
vaias concentraciones de resveratrol y en razón 2:3 de quercetina y ácido gálico con
respecto al resveratrol. Posteriormente se medirá la movilidad de los espermatozoides
mediante observación en cámara Makler, junto con su función mitocondrial mediante
citometría de flujo utilizando Rodamina 123 (R123), ioduro de propidio (PI) y diacetato
de 2'7'-didorofluoresceina (DCFH-DA), como marcadores de función mitocondrial ,
muerte celular y cantidad de ROS respectivamente. Finalmente se realizará un test de
zona-free (SPA) para evaluar la capacidad fecundante de los espermatozoides tratados.
Para este experimento se utilizarán ovocitos de hámster dorado (Mesocricetus auratus) ,
los cuales son despojados de la zona pelúcida. Se espera observar un aumento en la
movilidad, tanto en muestras normo como astenozoospérmicas expresado como el
aumento del porcentaje de espermatozoides móviles progresivos, además de un
aumento en la actividad mitocondrial expresado por una disminución de la fluorescencia
de R123. Finalmente se espera un aumento de penetraciones de los espermatozoides
en el SPA indicando una mayor capacidad fecundante. Los antioxidantes provenientes
de material de desecho pueden ser un valioso producto para la mejora de la tasa de
éxito de la IIU.
Currently, there has been an alarming dedine in semen quality in the world , which is directly affecting the pregnancy . This decline, in semen quality is probably ca u sed by human lifestyle, which leads to the generation of free radicals (ROS). These substances are highly reactive, and have a great affinity for membranes, making the ROS real cell killers. To combat these free radicals is natural to think on the use of antioxidants. These are substances capable of reduce the intracellular amount of free radicals, and thus help in the treatment against diseases and problems of everyday, reducing them. Our hypothesis is, the use of resveratrol , quercetin and gallic acid, antioxidants compounds from the grapevine (Vitis vinífera) , improves progressive mobility, mitochondrial function and fertilizing capacity of sperm samples from normal patients (normozoospermic) and on samples abnormal (asthenozoospermic). To prove this, we implicate a mixed solution of antioxidants at severa! concentrations of resveratrol and quercetin and gallic acid in a ratio of 2:3 relative to resveratrol. Then, we will measure mobility by observing on Makler chamber, along with mitochondrial function , using flow cytometry whit Rhodamine 123 (R123), propidium iodide (PI) and 2'7'-dichlorofluorescein diacetate (DCFH-DA), as markers of mitochondrial function, cell death and amount of ROS respectively. Finally there will procedure a zona-free test (SPA) to measure the fertilizing capacity of treated sperm. For this experiment we used golden hámster eggs (Mesocricetus auratus), which are removed of their pellucid zone. We expect to observe an increase in mobility in both norma as astenozoospérmic samples expressed as the increase of sperm progressive motile percentage, and a increase in mitochondrial activity expressed by a decrease in R123 fluorescence . Finally we expect observe an increase penetration of the sperm in the SPA indicating better fertilizing capacity. Antioxidants from the waste material can be a valuable product to improve the success rate of IUI.
Currently, there has been an alarming dedine in semen quality in the world , which is directly affecting the pregnancy . This decline, in semen quality is probably ca u sed by human lifestyle, which leads to the generation of free radicals (ROS). These substances are highly reactive, and have a great affinity for membranes, making the ROS real cell killers. To combat these free radicals is natural to think on the use of antioxidants. These are substances capable of reduce the intracellular amount of free radicals, and thus help in the treatment against diseases and problems of everyday, reducing them. Our hypothesis is, the use of resveratrol , quercetin and gallic acid, antioxidants compounds from the grapevine (Vitis vinífera) , improves progressive mobility, mitochondrial function and fertilizing capacity of sperm samples from normal patients (normozoospermic) and on samples abnormal (asthenozoospermic). To prove this, we implicate a mixed solution of antioxidants at severa! concentrations of resveratrol and quercetin and gallic acid in a ratio of 2:3 relative to resveratrol. Then, we will measure mobility by observing on Makler chamber, along with mitochondrial function , using flow cytometry whit Rhodamine 123 (R123), propidium iodide (PI) and 2'7'-dichlorofluorescein diacetate (DCFH-DA), as markers of mitochondrial function, cell death and amount of ROS respectively. Finally there will procedure a zona-free test (SPA) to measure the fertilizing capacity of treated sperm. For this experiment we used golden hámster eggs (Mesocricetus auratus), which are removed of their pellucid zone. We expect to observe an increase in mobility in both norma as astenozoospérmic samples expressed as the increase of sperm progressive motile percentage, and a increase in mitochondrial activity expressed by a decrease in R123 fluorescence . Finally we expect observe an increase penetration of the sperm in the SPA indicating better fertilizing capacity. Antioxidants from the waste material can be a valuable product to improve the success rate of IUI.
Notas
Tesis (Ingeniero en Biotecnología)
Palabras clave
Reproducción Humana, Espermatozoides, Antioxidantes, Uso Terapéutico