Effects of isoflavones on the skin of postmenopausal women: a pilot study

OBJECTIVE: The aim of this study was to evaluate the effects of isoflavones on the skin of postmenopausal women. DESIGN: A prospective study was performed with 30 postmenopausal women before and immediately after the end of treatment with 100 mg/day of an isoflavones-rich, concentrated soy extract for six months. A skin punch was performed in the gluteal region for sample collection before and immediately after the treatment program. Morphometric determination of epidermal thickness, the papillary index (wrinkling), and the amount of dermal elastic and collagen fibers was assessed. In addition, the number of blood vessels in the sample was also evaluated. The paired Student's t-test was used for statistical analysis (P < 0.05). RESULTS: Isoflavone treatment resulted in a 9.46% increase in the thickness of the epidermis in 23 patients. In addition, the papillary index was reduced in 21 women. The papillary index was inversely proportional to skin wrinkling, i.e., there were a large number of papillae after treatment. The amount of collagen in the dermis was increased in 25 women (86.2%). In 22 women (75.8%) we observed that the number of elastic fibers increased. The number of dermal blood vessels was significantly increased in 21 women. CONCLUSION: Our data show that the use of a concentrated, isoflavone-rich soy extract during six consecutive months caused significant increases in epithelial thickness, the number of elastic and collagen fibers, as well as the blood vessels.Federal University of São Paulo Gynecology DepartmentFederal University of São Paulo Morphology DepartmentUniversidade de São Paulo Faculdade de Medicina Laboratório de Ginecologia Estrutural e MolecularUniversidade de São Paulo Faculdade de Medicina Departamento de Obstetrícia e GinecologiaUNIFESP, Gynecology DepartmentUNIFESP, Morphology DepartmentSciEL

Effects of combined estrogen and raloxifene therapy on rat endometrium

PURPOSE: to evaluate the effects of conjugated equine estrogens (CEE) and raloxifene (Ral), alone or combined, on the rat endometrium. METHODS: fifty-six adult rats were ovariectomized and randomly divided into seven groups: GCont (control); GCEE (CEE 50 µg/kg); GCEE/25 (CEE 25 µg/kg); GRal/0.75 (Ral 0.75 mg/kg); GRal/0.4 (Ral 0.4 mg/kg); GCEERal (50/0.75) - (CEE 50 µg/kg + Ral 0.75 mg/kg), and GCEE-Ral (25/0.4) - (CEE 25 µg/kg + Ral 0.4 mg/kg). The drugs were orally administered (gavage) for 21 consecutive days. At the end of the experiment, all animals were anesthetized and sacrificed. Fragments of uterus were removed, fixed in 10% formaldehyde and processed for paraffin inclusion. The histological sections were stained by HE and submitted to histomorphometric evaluation. The following parameters were analyzed: thickness of superficial epithelium and number of endometrial glands/mm² and of blood vessels/mm². The data were evaluated using ANOVA followed by the Turkey-Kramer test. RESULTS: in the GCont and only Ral treatment (GRal/0.75 and GRal/0.4) the endometrium showed signals of atrophy. In the groups treated with only CEE signs of endometrial proliferation were observed, mainly in group GCEE/50. Also, there was endometrial proliferation in the groups that received combined CEE and Ral (Ral GCEE (50/0.75) and GCEE-Ral (25/0.4)), but it was more intensive in the animals treated with isolated estrogen than in those that received combined estrogen and raloxifene. CONCLUSION: raloxifene may partially block the action of estrogen on the castrated adult rat endometrium.OBJETIVO: avaliar os efeitos dos estrogênios conjugados eqüinos (Ece) e do raloxifeno (Ral), isoladamente ou associados, sobre o endométrio de ratas adultas. MÉTODOS: foram utilizadas 56 ratas adultas, ooforectomizadas, divididas aleatoriamente em sete grupos: GCont (controle); GEce (Ece 50 µg/kg); GEce/25 (Ece 25 µg/kg); GRal/0,75 (Ral 0,75 µg/kg); GRal/0,4 (Ral 0,4 mg/kg); GEce-Ral (50/0,75) - (Ece 50 µg/kg + Ral 0,75 mg/kg) e GEce-Ral (25/0,4) - (Ece 25 µg/kg + Ral 0,4 mg/kg). As drogas foram administradas por gavagem durante 21 dias consecutivos. Ao final da administração todos os animais foram anestesiados e fragmentos dos úteros removidos, fixados em formol a 10% e processados para inclusão em parafina. Os cortes obtidos foram corados por HE e submetidos à avaliação histomorfométrica. Foram avaliados os seguintes parâmetros: espessura do epitélio superficial, número de glândulas endometriais/mm² e número de vasos sangüíneos presentes no miométrio/mm². Os dados obtidos foram submetidos a análise estatística de ANOVA seguida pelo teste de Tukey-Kramer. RESULTADOS: nos grupos controle (Gcont) e tratados isoladamente com raloxifeno (GRal/0,75 e GRal/0,4) o endométrio mostrou-se atrófico. Já nos grupos tratados com Ece isoladamente foi observado endométrio muito desenvolvido, sendo este efeito mais acentuado no grupo que recebeu 50 µg/kg, no qual encontramos aumento da espessura do epitélio superficial e da lâmina própria e no número de glândulas endometriais e de vasos sanguíneos. Notamos também proliferação endometrial nos grupos que receberam a associação de Ece e Ral (GEce-Ral - 50/0,75) e (GEce-Ral - 25/0,4). CONCLUSÃO: o raloxifeno parece bloquear parcialmente a ação do estrogênio no endométrio de ratas adultas castradas.Universidade Federal de São Paulo (UNIFESP) Departamento de Ginecologia e ObstetríciaUniversidade Federal de São Paulo (UNIFESP)Universidade Federal de São Paulo (UNIFESP) Departamento de MorfologiaUniversidade Federal de São Paulo (UNIFESP) Departamento de GinecologiaUNIFESP, Depto. de Ginecologia e ObstetríciaUNIFESP, Depto. de MorfologiaUNIFESP, Depto. de GinecologiaSciEL

Proliferation of the superficial epithelium of ovaries in senile female rats following oral administration of conjugated equine estrogens

OBJECTIVE: To evaluate the effect of different concentrations of estrogen on the ovarian superficial epithelium in senile female rats. Design: Fifty female rats at 15 months of age and with irregular estrous cycles were selected and randomly divided into five experimental groups containing equal numbers of animals in each: GPROP, control group receiving vehicle only; GE0.05mg, group receiving conjugated equine estrogens (CEE) at a dose of 50 µg/kg; GE0.5mg, group receiving CEE at 500 µg/kg; GE1mg, group receiving CEE at 1 mg/kg; and GE2mg, receiving CEE at 2 mg/kg. The length of treatment was 21 days. After this period, the animals were anesthetized and the ovaries were fixed in 10% formaldehyde and processed for routine histology. Histomorphology was analyzed by light microscopy, and histomorphometrics were evaluated using the Imagelab program. RESULTS: In the GPROP and GE0.05mg groups, the superficial epithelium of the ovary had a simple cuboidal shape, and as the estrogen dose increased, the epithelium thickened, with pseudo-stratified or stratified epithelium appearing in the GE2mg group. The animals in the group given the highest estrogen dose (GE2mg) showed the thickest ovarian epithelium and the largest perimeter and surface area of the surface ovarian epithelium (P < 0.01). However, the difference in epithelium thickness between the GE0.5mg and GE1mg groups was only slight. CONCLUSION: Our data suggest that CEE at a dose of 2 mg/kg may induce marked proliferation of rat ovarian epithelium.UNIFESP Morphology Department Histology and Cellular Biology DivisionUniversidade de São Paulo Faculdade de MedicinaUniversidade Federal de São Paulo (UNIFESP) Faculdade de Medicina Gynecology DepartmentUNIFESP, Morphology Department Histology and Cellular Biology DivisionUNIFESP, Faculdade de Medicina Gynecology DepartmentSciEL

Seminal quality, testicle and epididymis morphology of bulls fed a diet containing gossypol

The objective was to evaluate the effect of intake of free gossypol on sperm quality and morphology of the testicles and epididymis of Nelore bulls. Twelve bulls were fed diets containing 3.3g of free gossypol/bull/day (cottonseed) (Group 1, n = 6) and a diet free of gossypol (Group 2, n = 6), respectively. Semen samples were collected in the beginning and end of the experiment which lasted 73 days. In the end of the study the testes and epididymis of bulls were removed to study the effect of free gossypol on histological characteristics. The average consumption of 3.3g of free gossypol/bull/day (mean 7.1mg of free gossypol/kg/day) reduced motility and sperm concentration and increased the percentage of major and total sperm defects, as well as the animals showing testes with seminiferous tubules of smaller thickness, fewer layers of spermatogenic lineage cells, smaller epididymal epithelium thickness and smaller number of sperm within the epididymal ducts, compared to animals with a diet free of gossypol (Group 2). The consumption of 3.3g of free gossypol/bull/day led to changes in morphology and morphometry of the testes and epididymis and reduced sperm quality of bulls.Objetivou-se avaliar o efeito da ingestão de gossipol livre sobre a qualidade espermática e a morfologia dos testículos e dos epidídimos de touros da raça Nelore. Doze touros receberam dieta contendo 3,3g de gossipol livre/touro/dia (caroço de algodão) (Grupo 1, n=6) e dieta isenta de gossipol (Grupo 2, n=6), respectivamente. Foram realizadas coletas de sêmen no início e no final do experimento, que teve duração de 73 dias. Ao final do estudo, foram retirados os testículos e os epidídimos dos touros para se estudar o efeito do gossipol livre sobre as características histológicas. O consumo médio de 3,3g de gossipol livre/touro/dia (média 7,1mg de gossipol livre/kg/dia) reduziu a motilidade e a concentração espermática e aumentou a porcentagem de defeitos espermáticos maiores e totais. Além disso, os animais apresentaram testículos com túbulos seminíferos de menor espessura de parede, menor número de camadas de células espermatogênicas, menor espessura do epitélio epididimário e menor número de espermatozoide no interior dos ductos epidídimários, em relação aos animais com dieta isenta de gossipol (Grupo 2). O consumo de 3,3g de gossipol livre/touro/dia acarretou alterações na morfologia e na morfometria dos testículos e dos epidídimos e reduziu a qualidade espermática dos touros.Universidade de Cuiabá Faculdade de Medicina VeterináriaUFMS Faculdade de Medicina Veterinária e ZootecniaEpamigUniversidade Federal do Mato GrossoUNIFESP-EPMUNIFESP, EPMSciEL

Glycosaminoglycan distribution in the rat uterine cervix during the estrous cycle

OBJECTIVE: To analyze the amount of glycosaminoglycans in the uterine cervix during each phase of the rat estrous cycle. DESIGN: Based on vaginal smears, forty female, regularly cycling rats were divided into four groups (n = 10 for each group): GI - proestrous, GII - estrous, GIII - metaestrous and GIV - diestrous. Animals were sacrificed at each phase of the cycle, and the cervix was immediately removed and submitted to biochemical extraction and determination of sulfated glycosaminoglycans and hyaluronic acid. The results were analyzed by ANOVA followed by the Bonferroni post-hoc test. RESULTS: The uterine cervix had the highest amount of total sulfated glycosaminoglycans and dermatan sulfate during the estrous phase (8.90 ± 0.55 mg/g of cetonic extract, p<0.001; and 8.86 ± 0.57 mg/g of cetonic extract, p<0.001). In addition, there was more heparan sulfate at the cervix during the proestrous phase (0.185 ± 0.03 mg/g of cetonic extract) than during any other phase (p<0.001). There were no significant changes in the concentration of hyaluronic acid in the uterine cervix during the estrous cycle. CONCLUSION: Our data suggest that the amount of total sulfated glycosaminoglycans may be influenced by hormonal fluctuations related to the estrous cycle, with dermatan sulfate and heparan sulfate being the glycosaminoglycans most sensitive to hormonal change.Universidade Federal de São Paulo (UNIFESP) Departmento de Morfologia e GenéticaUniversidade de São Paulo Faculdade de Medicina Departamento de Obstetrícia e GinecologiaUniversidade de São Paulo Instituto de Ciências Biomédicas Department of PharmacologyEscola Paulista de Medicina Department of GynecologyUNIFESP, Departmento de Morfologia e GenéticaEscola Paulista de Medicina Department of GynecologySciEL

Effects of high-dose isoflavones on rat uterus

OBJETIVO: Avaliar o efeito de altas doses de isoflavonas no útero de ratas adultas castradas. MÉTODOS: Ratas virgens ovariectomizadas (n = 40) foram tratadas por 30 dias consecutivos com veículo (GCtrl) ou genisteína nas concentrações 42 (GES42), 125 (GES125) e 250 (GES250) µg/g de peso corporal ao dia. O extrato de soja e o veículo (propilenoglicol) foram administrados por gavagem. Ao final do experimento, foi realizada dosagem sérica de 17 &#946;-estradiol e progesterona, avaliou-se o peso dos animais e dos úteros e foi feito exame colpocitológico. Fragmentos do terço médio dos cornos uterinos foram fixados em formol a 10% e processados para inclusão em parafina para estudo histológico. Cortes de 5 µm de espessura foram corados pelo HE e destinados a estudo em microscopia de luz. Analisou-se a histomorfologia do endométrio, área endometrial, número e área ocupada pelas glândulas, assim como a concentração de eosinófilos presentes na lâmina própria. Os dados numéricos obtidos foram submetidos à análise de variância complementada pelo teste de Tukey-Kramer (p < 0,05). RESULTADOS: Verificou-se que o peso do útero, as áreas endometrial e glandular e o número de glândulas e de eosinófilos foram maiores nos animais dos grupos GES250 &gt; GES125 do que nos outros grupos (GES250 &gt; GES125 &gt; GES42 = GCtrl; p < 0,05). Os dados morfológicos mostraram proliferação endometrial nos grupos ES125 e ES250, que apresentavam endométrio mais desenvolvido que outros grupos. Em todos os animais do grupo ES250 notou-se a presença de metaplasia escamosa. CONCLUSÃO: A administração de isoflavonas em altas doses promove metaplasia escamosa no endométrio.OBJECTIVE: To evaluate the effects of high-dose isoflavones on the uterus of castrated adult rats. METHODS: Adult, ovariectomized virgin rats (n = 40) were treated by gavage during 30 consecutive days with vehicle (propylene glycol, group GCtrl) or different doses of genistein: 42 (group GES42), 125 (GES125), or 250 (GES250) µg/g body weight per day. Animals were killed, weighed, vaginal and uterine samples were taken for cytologic evaluation, and serum levels of 17 &#946;-estradiol and progesterone were determined. The middle third of the uterine horns was dissected, fixed in 10% formaldehyde and processed for paraffin inclusion; 5-µm thick sections were obtained and stained with HE for further histological study under light microscopy. The endometrial morphology and area, number and area of glands, and number of eosinophils in the lamina propria were analyzed. ANOVA and the Tukey-Kramer test were used for statistical analyses. RESULTS: Uterine weight, endometrial glandular area, and number of glands and eosinophils were all higher in GES250 &gt; G125 than in the other groups (GES250 &gt; GES125 &gt; GES42 = GCtrl; p < 0.05). Morphological data showed signs of endometrial proliferation upon treatment with genistein, especially in animals in GES125 and GES250 compared to other groups. In all animals in GES250, signs of uterine squamous metaplasia were observed. CONCLUSION: A short treatment period with high daily doses of isoflavones can promote endometrial squamous metaplasia in ovariectomized rats.CNPqCoordenacao de Aperfeicoamento de Pessoal de Nivel Superior (CAPES

Endometrium in women with polycystic ovary syndrome during the window of implantation

The human endometrium undergoes to a complex series of prolifertive and secretory changes in each menstrual cycle and displays only a short period of receptivity, known as the window of implantation, necessary for the implantation of the blastocyst in the uterus. The implantation process occurs in a sequential manner, leading to the establishment of pregnancy. Morphofunctional changes during this period may prevent or hinder the implantation. For this reason, the study of the endometrium at this stage is important for the improvement of therapies that may interfere with the mechanisms involved in maternal-embryonic interaction. Several gynecological disorders, including polycystic ovary syndrome (PCOS), are associated with decreased fertility and uterine receptivity. In spite of recent advances in assisted reproduction techniques, allowing the selection of high quality embryos, the implantation rate remains low and has not increased enough in recent decades. This article aims at reviewing the endometrial aspects of the window of implantation in women with polycystic ovary syndrome, focusing mainly on adhesion molecules. For that purpose, we analyzed 105 articles published in journals indexed in PubMed in the last 50 years (up to May 2011). In conclusion, the endometrial receptivity seems to be the major limiting factor for the establishment of pregnancy in a large number of gynecological diseases, including PCOS, and treatment to improve implantation rates is likely to be taken towards this direction.O endométrio humano é submetido a uma complexa série de mudanças proliferativas e secretórias em cada ciclo menstrual e exibe somente pequeno período de receptividade, conhecido como janela de implantação, necessário para a nidação do blastocisto no útero. O processo da implantação ocorre de forma sequencial, levando ao estabelecimento da gravidez. Alterações morfofuncionais durante este período podem impedir ou dificultar a implantação. Por este motivo, o estudo do endométrio nesta fase é importante para o aprimoramento de terapias que possam interferir nos mecanismos envolvidos na interção materno-embrionária. Várias doenças ginecológicas, incluindo a síndrome dos ovários policísticos (SOP), estão associadas à diminuição da fecundidade e da receptividade uterinas. Apesar de recentes avanços nas técnicas de reprodução assistida, permitindo a seleção de embriões de alta qualidade, a taxa de implantação continua baixa e não tem aumentado suficientemente nas últimas décadas. O presente artigo tem como objetivo revisar os aspectos endometriais da janela de implantação em mulheres com a síndrome dos ovários policísticos, focando especialmente as moléculas de adesão. Para tanto, nos valemos da análise de 105 artigos publicados em revistas indexadas no PUBMED nos últimos 50 anos (até maio de 2011). Como conclusão, a receptividade endometrial parece ser o maior fator limitante no estabelecimento da gestação em grande número de doenças ginecológicas, incluindo a SOP, e o tratamento para melhorar as taxas de implantação possivelmente será nessa direção.Universidade Federal de São Paulo (UNIFESP) Departamento de GinecologiaUniversidade Federal do PiauíUniversidade de São Paulo Faculdade de MedicinaUNIFESP Departamento de MorfologiaFMUSP Departamento de Obstetrícia e GinecologiaUNIFESP, Depto. de GinecologiaUNIFESP, Depto. de MorfologiaSciEL

Supplementation with fish oil reduces morphological aspects of muscle damage induced by intense exercise in rats = Suplementação com óleo de peixe reduz aspectos morfológicos de lesão muscular induzida por exercícios intensos em ratos

OBJETIVOS: Investigar os efeitos do exercício resistido e da ingestão de óleo de peixe na morfologia da fibra muscular em ratos Wistar. MÉTODOS: Quarenta e oito animais realizaram exercício resistido e foram divididos inicialmente em dois grupos. Um dos grupos não recebia óleo de peixe e o outro grupo ingeria o óleo de peixe. Os animais do segundo grupo realizaram o treinamento e ingeriram o óleo de peixe por um período de oito semanas. Ao final da última sessão de exercício resistido os animais foram organizados em seis subgrupos de oito cada, segundo o intervalo de tempo (12, 24 e 48 horas) transcorrido até o procedimento de retirada do músculo gastrocnêmio. Em cada tempo determinado após a última sessão de exercício resistido, o músculo gastrocnêmio foi retirado para análise morfológica. RESULTADOS: As células do músculo esquelético dos animais que não receberam óleo de peixe apresentaram escores maiores de edema, especialmente as dos grupos que tiveram os músculos retirados em 24 e 48 horas. No grupo que ingeriu o óleo de peixe observou-se menor quantidade de infiltrado inflamatório e áreas de necrose reduzidas em comparação com os animais que se exercitavam sem o uso de óleo de peixe, em todos os intervalos de tempo pós-exercício. CONCLUSÕES: A ingestão de óleo de peixe atenuou as alterações morfológicas no tecido muscular após exercícios de alta intensidad

Does Gestrinone Antagonize the Effects of Estrogen on Endometrial Implants Upon the Peritoneum of Rats?

OBJECTIVE: To evaluate the effects of estrogen treatment in combination with gestrinone on an experimental rat model of endometriosis. METHODS: Uterine transplants were attached to the peritoneum of female Wistar rats via a surgical autotransplantation technique. The implanted area was measured during the proestrus phase and after hormonal treatment. We performed morphometric analysis and examined the macroscopic and morphometric alterations of endometrial implants after hormonal treatment in ovariectomized rats. RESULTS: The high dose of estrogen caused macroscopic increases in the endometrial implant group compared with other groups, which were similar to increases in the proestrus phase. The low dose showed morphometric development of implants, such as an increase in number of endometrial glands, leukocyte infiltration and mitosis. Gestrinone antagonized both doses of estrogen. CONCLUSION: Our findings suggest that gestrinone antagonizes estrogen's effects on rat peritoneal endometrial implants

Influence of hyperprolactinemia on collagen fibers in the lacrimal gland of female mice

OBJECTIVE:To quantify the collagen fibers in the lacrimal gland of female mice with hyperprolactinemia.METHODS:Forty adult female mice were randomly divided into two groups with 20 animals each: nonpregnant control (CTR1, control group, 0.2 mL of saline solution) and nonpregnant experimental (HPRL1, experimental group, 200 µg/day metoclopramide). Treatments lasted for 50 consecutive days. On day 50, 10 females from each group (control and experimental) were euthanized in the proestrus phase; then, the blood was collected and the lacrimal glands were removed. Thereafter, the remaining females were placed with the mates and continued to receive treatment with saline solution or metoclopramide. On the 6th post-coital day, 10 pregnant females from the control group (CTR2) and 10 pregnant females from the experimental group (HPRL2) were euthanized, after which blood was collected and the lacrimal glands removed. The lacrimal glands were processed for morphological analyses and collagen quantification, and prolactin and sex steroid levels were measured in the blood samples. Data were statistically analyzed using an unpaired Student t test (