A fost validată o metodă simplă şi selectivă de determinare a genisteinei (GNST) din ficat şi rinichi de şobolan în vederea studierii efectului fitoestrogenic al GNST la șobolani Wistar de sex ...feminin ovariectomizaţi. GNST a fost separată pe o coloană Kromasil 100-RP8, de 150 mm x 4,6 mm, 5 μm prevăzută cu precoloană Kromasil RP 8. Faza mobilă a fost 55:45 (v/v) acid fosforic 15 mmol în apă : metanol la un debit de 1,3 ml/min. Ca standard intern a fost folosită luteolina 20 μg/ml în metanol. Timpul de retenţie al GNST a fost tR = 13,22 min şi tR = 11,60 min pentru standardul intern. Curbele de calibrare pe domeniul 40-400 μg GNST/100g ficat şi 20-200 μg GNST/100g rinichi au prezentat coeficienţi de determinare ˃0,99. Metoda a prezentat o bună precizie şi acurateţe la limita de cuantificare. 10 şobolani albi Wistar de sex feminin, cu vârsta de 8 săptămâni au fost trataţi s.c. cu 10 mg/kg c/zi soluţie GNST timp de 8 săptămâni, în timp ce un lot de 10 animale au folosit ca martor. Valorile obţinute pentru GNST în ficat au fost 192.12 ± 53.46 μg/100g, iar în rinichi 74.51 ± 12.77 μg/100g.
A simple and selective method for genistein (GNST) determination in rat liver and kidney was validated in order to study the phytoestrogenic effect of GNST in ovariectomised female Wistar rats. GNST ...was separated on a Kromasil 100-RP8 column, 150 mm x 4.6 mm, 5 mm equipped with a Kromasil RP 8 precolumn. The mobile phase was 55:45 (v / v) phosphoric acid, 15 mmol in water: methanol at a flow rate of 1.3 ml / min. Luteolin 20 μg / ml in methanol was used as internal standard (IS). The retention time of GNST was t
= 13.22 min and t
=11.60 min for the IS. Calibration curves in the range 40-400 μg GNST/100g liver and 20-200 μg GNST/100g kidney presented a coefficient of determination higher than 0.99. The method developed presented a good precision and accuracy at the lower limit of quantification LLOQ. 10 white Wistar female rats, 8 weeks of age were treated s.c. with 10 mg GNST/kg bw/day for 8 weeks, while a group of 10 animals were used as controls. The values obtained for GNST in the liver were 192.12 ± 53.46 μg/100g and 74.51 ± 12.77 μg/100g in kidney samples.
A fost validată o metodă simplă şi selectivă de determinare a genisteinei (GNST) din ficat şi rinichi de şobolan în vederea studierii efectului fitoestrogenic al GNST la șobolani Wistar de sex feminin ovariectomizaţi. GNST a fost separată pe o coloană Kromasil 100-RP8, de 150 mm x 4,6 mm, 5 μm prevăzută cu precoloană Kromasil RP 8. Faza mobilă a fost 55:45 (v/v) acid fosforic 15 mmol în apă : metanol la un debit de 1,3 ml/min. Ca standard intern a fost folosită luteolina 20 μg/ml în metanol. Timpul de retenţie al GNST a fost t
= 13,22 min şi t
= 11,60 min pentru standardul intern. Curbele de calibrare pe domeniul 40-400 μg GNST/100g ficat şi 20-200 μg GNST/100g rinichi au prezentat coeficienţi de determinare ˃0,99. Metoda a prezentat o bună precizie şi acurateţe la limita de cuantificare. 10 şobolani albi Wistar de sex feminin, cu vârsta de 8 săptămâni au fost trataţi s.c. cu 10 mg/kg c/zi soluţie GNST timp de 8 săptămâni, în timp ce un lot de 10 animale au folosit ca martor. Valorile obţinute pentru GNST în ficat au fost 192.12 ± 53.46 μg/100g, iar în rinichi 74.51 ± 12.77 μg/100g.
Soy phytoestrogens, primarily genistein and daidzein, are widely used through diet and as dietary supplements. Since they may act as “natural” selective modulators of estrogen receptor, scientific ...interest for phytoestrogens is constantly increasing. Cessation of a women’s reproductive function, i.e. menopause, is often accompanied with disorders that are consequence of reduced ovarian function and low estrogen concentration. Therefore, one of the possible therapeutic application of genistein and daidzein is prevention and alleviation of menopausal symptoms, as an alternative to hormone replacement therapy. The aim of this study is to examine the effects of phytoestrogens on the reproductive system of immature and middle-aged female rats. Of particular interest is to define the potential of phytoestrogens for improvement of reproductive system function in an animal model of menopause, and to compare these effects with the effects of therapeutic doses of estradiol dipropionate, commonly used in prevention and treatment of menopausal symptoms. Uterotrophic assay was used in order to test estrogenic activity of genistein and daidzein, according to the protocol of the Organization for Economic Cooperation and Development (OECD). Immature female rats received phytoestrogens (genistein and daidzein), in three doses (high, medium and low) for three consecutive days, from 19th till 21st day of life. According to the protocol, control groups were formed - the positive control groups that received 17α - ethynyl estradiol (in two doses), negative control groups that received solvents, and the intact control group. After uterotrophyc assay, minimal dose of genistein and daidzein that exerts estrogenic effect was determined. This dose was administered to 12 months old, middleaged female rats, for four weeks. After this period it was analyzed whether genistein and daidzein exerted beneficial effects on the structure and function of the pituitary, ovary, uterus and vagina. These effects were then compared with the effects of the therapeutic dose of estradiol dipropionate. Changes in the structure and function of the organs of reproductive system of immature and middle-aged female rats were analyzed by histological, immunofluorescence, design-based stereological, morphometric and molecular biological methods. Immunofluorescent labeling of specific antigens allowed the visualization of the pituitary lactotropic and gonadotropic cells, and the calculation of the relative intensity of fluorescent signal, i.e. concentration of antigen within the cell. In the same manner, the presence of lactoferrin and steroid receptors in the uterus of immature and middle-aged female rats was analysed. Unbiased, design-based stereological methods were used for determination of volume of reproductive organs, volume density of their basic compartments, as well as volume, number and volume density of granulosa lutein cells of the corpora lutea. The endometrial epithelium height, endometrial gland number, number of ovarian follicles at different stages of folliculogenesis and atresia, as well as the number of individual classes and the total number of corpora lutea were performed using morphometric methods. "Real-time" PCR was used to determine the relative gene expression of estrogen receptor alpha (ERα), estrogen receptor beta (ERβ) and progesterone receptor (PR) in the uterus of middle-aged female rats. All results were statistically analyzed. Uterotrophic assay showed that genistein, at doses of 35 and 50 mg/kg b.w., increased gravimetric uterine growth and acted as an estrogen agonist, while daidzein, in any dose used, did not cause these effects. It has been shown that genistein, in medium and high dose, acted as an estrogen agonist in terms of all tested parameters in uterus, pituitary gland, and vagina. However, the same doses in the ovaries caused antagonistic effects. By including additional analyses, the sensitivity of uterotrophic assay was increased. This resulted in an observation that daidzein also exerts estrogenic activity in all tested doses. The estrogenic activity of daidzein was reflected in the increase of endometrial gland number and volume density, and stimulation of pituitary gonadotropic and lactotropic cells. The minimal effective dose, i.e. the dose that causes the estrogenic effect in the organs of female reproductive system, is the dose of 35 mg/kg b.w. per day of genistein and daidzein. Chronic estradiol treatment of middle-aged female rats caused an inhibition of gonadotropic cells, which was reflected not only in the reduction of fluorescent signal intensity, but also in reduction of the cell size. At the same time, estradiol stimulated lactotrops, by increasing their density and the relative intensity of the immunolabeled prolactin in cells. In contrast to estradiol treatment, genistein and daidzein did not cause changes in the relative intensity of the fluorescent signal within the gonadotropic or lactotropic cells. However, changes in cell morphology were observed after treatment with both phytoestrogens. Gonadotropic cells were larger in size, while lactotropic cells were smaller comparing to the controls. All types of examined pituitary cells were changed in shape with unevenly stained cytoplasm, that is, immunolabeled parts of cytoplasm were separated by unstained regions, which gave cells the appearance of vacuolization. Chronic treatment with therapeutic doses of estradiol, in the ovaries of acyclic females, generally had a negative effect. In the group of small follicles, estradiol caused a significant increase of atretic, primordial, primary and preantral follicles. Similar effect was observed in the group of antral follicles. Estradiol treatment caused an increase of volume and number of corpora lutea, which resulted in the ovary volume increase. In contrast to estradiol, chronic application of genistein and daidzein caused a significant increase in the number of follicles in initial stage of folliculogenesis (primordial and primary follicles), without affecting total number of atretic primordial follicles, but significantly reducing the number of primary atretic follicles. Treatments with phytoestrogens did not affect total number of preantral and antral follicles, while they caused a decrease of atretic antral follicle number. Genistein treatment did not change the number of individual classes of corpora lutea, or their total number. Unlike genistein, daidzein increased the number of mature corpora lutea, which resulted in an increase of their total number, and ovary volume. However, due to the preservation of healthy follicles, both treatments caused an increase of follicular parenchyma volume. Estradiol treatment increased the absolute and the relative uterine weights. This is result of increased volume density of endometrium, endometrial epithelium and endometrial glands. Estradiol caused decrease of ERα mRNA and increase of PR mRNA expression, while ERβ mRNA expression was not changed. Genistein caused an increase of uterine weight and endometrial volume density, decrease of ERα mRNA expression, and increase of PR and ERβ mRNA expression. In cotrast to genistein treatment, daidzein did not change uterine weight, or morphometric and stereological characteristics of uterus. Expression of ERα and PR mRNA was not affected, while expression of ERβ mRNA was increased. In vagina, application of estradiol caused hypertrophy of the epithelium, followed by desquamation of epithelial cells. After treatment with phytoestrogens, changes in general histological appearance of the vaginal epithelium were not observed. However, detailed light microscopic analysis revealed less numerous pyknotic nuclei, and more numerous mitotic cells. Comparing to estradiol, genistein and daidzein, exhibited numerous beneficial effects on the reproductive system of middle-aged females, especially in the ovary. Bearing in mind that menopause, among other things, is caused by reduced ovarian function, the results of this study are significant and applicable, given that the preservation of ovarian function can significantly improve the quality of life of women. Therefore, it can be concluded that genistein and daidzein could be a successful alternative to estrogen replacement hormone therapy.
Fitoestrogeni iz soje, pre svega genistein i daidzein, su veoma zastupljeni u životu savremenog čoveka, kako kroz ishranu, tako i kroz pomoćna lekovita sredstva. Zbog svog selektivno modulatorskog, tj. tkivno specifičnog estrogenskog delovanja, fitoestrogeni poslednjih godina postaju atraktivno polje intenzivnih studija. Period gašenja reproduktivne funkicije kod žena, tj. menopauza, vrlo često je praćen tegobama i poremećajima koje su posledica smanjenja koncentracije endogenog estradiola, odnosno funkcije ovarijuma. Stoga je jedna od mogućih terapijskih aplikacija genisteina i daidzeina upravo sprečavanje i ublažavanje ovih tegoba, a kao alternativa supstitucionoj hormonskoj terapiji. Međutim, njihovo delovanje na organe ženskog reproduktivnog sistema, naročito u periodu menopauze, još uvek nije u potpunosti istraženo. Cilj ove studije obuhvata rasvetljavanje dejstava fitoestrogena na reproduktivni sistem nezrelih i acikličnih ženki pacova. Od posebnog interesa je jasno definisanje potencijala fitoestrogena u poboljšanju funkcije reproduktivnog sistema u animalnom modelu menopauze, kao i poređenje efekata njihove aplikacije sa onima koje ima aplikacija terapijske doze estradiol dipropionata, koji se koristi kao lek izbora u prevenciji i terapiji menopauzalnih simptoma. Za ispitivanje estrogenske aktivnosti fitoestrogena (genisteina i daidzena) korišćen je uterotrofni esej po protokolu Organizacije za ekonomsku saradnju i razvoj (OECD). Ženkama pacova starosti 19 dana aplikovani su fitoestrogeni (genistein i daidzein), tokom tri dana u tri doze (visokoj, srednjoj i niskoj). U okviru protokola predviđene su i kontrolne grupe - pozitivne kontrolne
Objectivo: Avaliar a eficácia dos fitoestrogénios no tratamento dos sintomas vasomotores da peri e pós-menopausa. Fontes de dados: UptoDate, MEDLINE, Cochrane Library, Bandolier, DARE, TRIP, National ...Guideline Clearinghouse, Índex de Revistas Médicas Portuguesas e revistas especializadas. Métodos de revisão: Pesquisa bibliográfica utilizando os termos MeSH phytoestrogens, perimenopause, postmenopause e hot flashes, publicados entre Janeiro de 2000 e Janeiro de 2011, em Inglês e Português. Foi utilizada a Strength of Recommendation Taxonomy (SORT) para avaliação da qualidade dos estudos e posterior atribuição de níveis de evidência e forças de recomendação. Resultados: Dos 78 artigos identificados, foram seleccionados 14 (um sistema, quatro normas de orientação clínica, três meta-análises, quatro revisões sistemáticas e dois ensaios clínicos aleatorizados controlados). A recomendação do Uptodate e uma meta-análise publicada na Cochrane (nível de evidência 1) concluem não haver eficácia dos fitoestrogénios. Os resultados dos restantes artigos analisados, na generalidade com nível de evidência 2, são conflituosos e a maioria demonstra não haver benefício com o uso dos fitoestrogénios na redução da severidade e frequência dos sintomas vasomotores da peri e pós-menopausa. Conclusões: Da revisão da evidência encontrada, o uso dos fitoestrogénios parece ser ineficaz na redução dos sintomas vasomotores da peri e pós-menopausa (Força de Recomendação A). Denotam-se diferenças entre os desenhos dos estudos, parti-cularmente no tipo e dose de composto avaliado.
Objetivo: avaliar os efeitos estrogênicos de fitoestrogênios da soja em pacientes na pós-menopausa. Métodos: foram avaliadas pacientes na pós-menopausa pré-selecionadas no Setor de Climatério do ...Departamento de Ginecologia da Universidade Federal de São Paulo- Escola Paulista de Medicina. As pacientes foram divididas em dois grupos de 40 cada: GI (isoflavona) - recebeu 100 mg de isoflavona por dia, em cápsulas (divididas em 3 tomadas); e GCrt - recebeu placebo (controle). Na inclusão do trabalho, foram avaliadas por meio do índice de Kupperman, exame físico e submetidas a dosagens bioquímicas de colesterol total e frações, triglicérides, estradiol, FSH e LH, além de avaliação ultra-sonográfica do eco endometrial. Para comparar os dois grupos antes e após o tratamento, utilizou-se o teste não paramétrico de Kruskal-Wallis para o índice de Kupperman, ao passo que os dados foram analisados pelo teste t para duas amostras independentes. Resultados: no que se refere aos sintomas avaliados pelo índice de Kupperman, 80 % das pacientes do GI (isoflavona) melhoraram, ao passo que no GCtr (controle) a melhora foi de apenas 12,5%. Com relação aos níveis de colesterol total plasmático, das 35 pacientes do GI que apresentaram níveis superiores a 200 mg/dL, ao passo que a queda registrada no GCtr foi em apenas 13 pacientes. Com relação ao índice de massa corpórea, houve redução no grupo GI (isoflavona), mantendo-se estável no GCtr (controle). Notamos também que a espessura endometrial e os níveis sérios de FSH não apresentaram variações significativas. Houve elevação dos valores de estrogênio após o tratamento com isoflavona. Conclusão: concluímos que a isoflavona, na dose de 100 mg/dia, pode ser utilizada como alternativa para atenuar os sintomas climatéricos. Comprovamos que houve redução dos níveis de colesterol total plasmático.
Oral exposure to synthetic and natural endocrine disruptors (ED) at different stages of life is related to alterations in the reproductive system. Ruminants represent a model for studying the effects ...of ED in humans due to some similarities in their development. Ruminants are considered of great importance due to continuous exposure to pasture and contaminated soils and human consumption of their meat. The present review aims to describe ED effects in ruminants’ reproduction, when they are consumed in the perinatal, postnatal-pre-adulthood, and adult stages. Feeding with phytoestrogens (PE) rich forage has no nocive consequences on the male reproductive system at any stages of life; in females, they produce more significant harmful effects when administered in adulthood. In the perinatal and postnatal-pre-adulthood stages, negative effects of synthetic ED were found in both males and females. This review presents study opportunities for further research related to oral exposure to ED.
La exposición oral a disruptores endocrinos (DE) sintéticos y naturales en las distintas etapas de la vida está relacionada con alteraciones en el sistema reproductivo. Los rumiantes representan modelos adecuados para estudiar el impacto de los DE en los humanos por algunas similitudes en su desarrollo, se consideran de gran importancia por la exposición continua a pastura y suelo contaminado, y por el consumo humano de su carne. El objetivo de la presente revisión es describir los efectos producidos por consumir DE en etapa perinatal, posnatal-preadultez y adultez en rumiantes. La alimentación con forraje rico en fitoestrógenos (FE), no tiene efectos nocivos sobre la reproducción de machos en ninguna etapa de la vida; en las hembras, producen mayores efectos nocivos cuando se administran en la adultez. En la etapa perinatal y posnatal-preadultez, se encontraron efectos negativos de los DE sintéticos, tanto en machos como en hembras. La presente revisión da a conocer las oportunidades de estudios para continuar con investigaciones relacionadas a la exposición oral a DE.
Los fitoestrógenos (FEs) son compuestos químicos provenientes del metabolismo secundario de algunos vegetales, tienen un efecto potencial sobre los parámetros reproductivos de los animales ...domésticos, al actuar como agonistas o antagonistas de los receptores estrogénicos. El objetivo de esta revisión es conocer los efectos que produce una dieta rica en FEs en la fisiología reproductiva de los animales de abasto. Se realizó una revisión sistemática en dos bases de datos mediante el uso de palabras clave relacionadas con los efectos que produce la ingestión de FEs en la dieta, sobre la reproducción de animales de abasto, únicamente se consideraron los estudios controlados desarrollados in vivo. Se encontraron resultados contradictorios, por un lado, la ingesta de un alto contenido de los compuestos polifenólicos, provenientes de distintos forrajes en la hembra bovina, estuvo relacionado con la disminución de la fertilidad, presencia de abortos y de quistes ováricos; por otro lado, la ingesta de alto contenido de FEs indujo un aumento en la calidad seminal de los machos de las especies: bovinos, ovinos y lepóridos; por lo que, dichos efectos se pueden atribuir a la concentración, al tipo de FEs, sexo, especie e incluso a la raza del animal.
Alimentos funcionales, alfalfa y fitoestrógenos Alejandro De Jesús Cortés-Sánchez; José Rubén León-Sánchez; Francisco Javier Jiménez-González ...
Mutis- Revista de arte y ciencia de la Universidad Jorge Tadeo Lozano,
05/2016, Volume:
6, Issue:
1
Journal Article
Peer reviewed
Open access
El desarrollo de alimentos funcionales en los últimos años ha ido en aumento debido principalmente, al interés de la población por alimentos que no solo cumplan con la función nutricional sino ...también por que tengan un efecto benéfico en salud reduciendo así el riesgo a padecer diversas enfermedades. La alfalfa ha sido generalmente un alimento destinado para la alimentación del ganado, sin embargo, en diversas investigaciones alrededor del mundo se ha reportado que contiene compuestos bioactivos –fitoquímicos– y proteínas con efecto estrogénico, antimicrobiano y antioxidante, así como también protector contra enfermedades cardiovasculares, cáncer y diabetes, entre otras, clasificándola potencialmente como un alimento funcional. El presente documento tiene como objetivo dar un bosquejo general informativo a través de la búsqueda y revisión bibliográfica acerca de los alimentos funcionales y en especial de la alfalfa y constituyentes fitoquímicos (fitoestrógenos) que la ubican dentro de este grupo de alimentos; así como los hallazgos que se han reportado como efectos adversos en animales experimentales específicamente en el aparato reproductivo debido al consumo de este tipo de fitocompuestos. Esta dicotomía entre los efectos adversos y benéficos no es más que la señalización de que aún falta mucha más información y estudios que realizar respecto al resultado que pueden tener los fitoestrógenos en el desarrollo y función reproductiva de animales, el agonismo y antagonismo de su efecto así como la concordancia en el riesgo-beneficio de su consumo.
In this article, we describe the physicochemical characteristics of those flavonoid compounds which have been more deeply studied up to the present. We also analyze their contribution to traditional ...and natural medicine, together with their applications in food industry. The main objectives are, on the one hand, to provide basic physicochemical information about the molecular structures of the kinds which have more industrial and medical applications up to the present, and, on the other hand, to connect the natural properties of the foods that contain them with their benefits to human health.
En el presente trabajo se describen las características fisicoquímicas de los compuestos Flavonoides con mayor grado de estudio hasta la actualidad, y su aporte a la medicina tradicional y natural, junto con sus aplicaciones en el campo de la industria alimenticia. Los principales objetivos del mismo son, por un lado, proveer información fisicoquímica básica sobre las estructuras moleculares de los diferentes tipos con mayores aplicaciones industriales y medicinales hasta el presente, y por otro, relacionar las propiedades naturales de los alimentos que los contienen con los beneficios a la salud humana.
Las isoflavonas de soya son un grupo de compuestos bioactivos, no nutritivos, de naturaleza fenólica, no esteroidal, encontrados en la soya y alimentos derivados. Comparten con otros compuestos la ...capacidad de unión a los receptores estrogénicos de distintas células y tejidos, por lo que pueden actuar como fitoestrógenos. El interés actual sobre ellas deriva del reconocimiento de que poblaciones asiáticas con un alto consumo presentan menor prevalencia de cáncer y enfermedades cardiovasculares, en comparación con poblaciones de países occidentales. Este beneficio cardiovascular no estaría dado sólo mediante modulación de los lípidos plasmáticos, mecanismo ampliamente estudiado. En este artículo se revisa la evidencia publicada acerca de los efectos benéficos de las isoflavonas de soya y las distintas vías de acción que beneficiarían la salud cardiovascular, mecanismos que trascienden los tradicionalmente abordados, como la modulación de los lípidos plasmáticos, y que involucran regulación de funciones a nivel celular y enzimático, en eventos como la inflamación, trombosis y progresión aterosclerótica.
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