Imbalance between the cellular proliferation and death in harderian gland of female mice with hyperprolactinemia.

OBJECTIVE: To assess the impact of the metoclopramide-induced hyperprolactinemia in cellular death and proliferation in the harderian gland of female mice. METHODS: Twenty female mice were divided into two groups of 10 animals each and treated: 0.2 mL of saline solution (controls, Ctr) and 200 µg of metoclopramide (experimental, hyperprolactinemia), both for 50 consecutive days and at 12:00 a.m. On the 50th day, the female were euthanized, and the harderian glands were removed and processed for immunohistochemistry for detected ki67 and TUNEL method. Data were statistically analyzed by unpaired Student's t test (p<0.05). RESULTS: The harderian gland of the hyperprolactinemia group showed increase in the immunoexpression of Ki67 and TUNEL compared to the Ctr group (p<0.05), and there was no significant difference in the amount of porphyrin in the HPrl group compared to the Ctr group. CONCLUSION: The hyperprolactinemia led to increased cell death in the acini the harderian gland and cell proliferation in the stroma glandular, fact that suggesting a reduction process of cellular activity and fibrosis, which suggests impairment in the functioning of the lacrimal harderian.

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 (p<0.05). RESULTS: Morphological analysis revealed greater structural tissue disorganization of the lacrimal glands in the metoclopramide-treated groups. The total collagen content was significantly higher in the HPRL1 group than in the CTR1 group (p<0.05), whereas the difference between the CTR2 and HPRL2 groups was not significant. CONCLUSION: Our data suggest an impairment in the functioning of the lacrimal gland as a consequence of increased prolactin levels and decreased serum levels of estrogen and progesterone.

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 (p<0.05).RESULTS:Morphological analysis revealed greater structural tissue disorganization of the lacrimal glands in the metoclopramide-treated groups. The total collagen content was significantly higher in the HPRL1 group than in the CTR1 group (p<0.05), whereas the difference between the CTR2 and HPRL2 groups was not significant.CONCLUSION:Our data suggest an impairment in the functioning of the lacrimal gland as a consequence of increased prolactin levels and decreased serum levels of estrogen and progesterone.

The evaluation of endometrial sulfate glycosaminoglycans in women with polycystic ovary syndrome.

The aim of this study was to quantify the sulfated glycosaminoglycans in the endometria of women with polycystic ovary syndrome (PCOS). Of the 18 patients recruited for this study, 10 patients with PCOS comprised the PCOS group (PCOSG), and eight patients with regular and ovulatory menstrual cycles comprised the control group (CG). The clinical, biochemical, morphological and endometrial data from both groups were analyzed. Biopsies were performed during the proliferative phase of the menstrual cycle for the CG and during the persistent proliferative phase for the PCOSG (all women were amenorrheic). In the PCOSG, there was a significant increase in the endometrial concentration levels of heparan sulfate (p = 0.03), but no difference in the concentrations of chondroitin sulfate was determined between the two groups (p = 0.77). Period of time without menstruation (p = 0.001) and body mass index (BMI) (p = 0.04) correlated directly and positively with heparan sulfate concentration. There was no association between heparan sulfate levels and basal insulin values (p = 0.08). High levels of endometrial heparan sulfate in women with PCOS indicate an interference with maternal-fetal recognition, which contributes to infertility; thus, endometrial heparan sulfate may be a predictive marker of future neoplasia risk.

Hyperprolactinemia changes the sulfated glycosaminoglycan amount on the murine uterus during the estrous cycle.

OBJECTIVE: To evaluate the effects of hyperprolactinemia on the sulfated glycosaminoglycans (GAGs) of the murine uterus. DESIGN: Experimental research. SETTING: University biochemistry laboratory. ANIMAL(S): Eighty female mice were divided into two groups of 40 animals each and treated with 0.2 mL of saline solution (controls, Ctr) and 200 µg of metoclopramide (experimental, HPrl). Treatments lasted for 50 consecutive days. The animals were divided into four subgroups of 10 animals each per treatment (Ctr and HPrl) and sacrificed according to the phase of the estrous cycle. The uterine horns were removed for biochemical analyses, and blood samples were collected for hormone measurements. INTERVENTION(S): Induced hyperprolactinemia. MAIN OUTCOME MEASURE(S): To quantify the sulfated GAGs, and PRL and sex steroid levels. RESULT(S): The endometrium during the estrus phase was significantly thicker in the HPrl animals than in the Ctr mice. The levels of chondroitin and dermatan sulfate were significantly increased in the HPrl group than in the Ctr group during all phases except metestrus. The amounts of heparan sulfate were lower during estrus and diestrus and higher in the metestrus phase in HPrl than in Ctr animals. Serum PRL levels were increased whereas the levels of E2 and P were decreased in all phases in the HPrl group than in the Ctr group. CONCLUSION(S): The hyperprolactinemia changed the amounts of uterine sulfated GAGs. Our data suggest that these changes may not be correlated with ovarian steroid levels.

[Morphological aspects of the urethra in female rats after electrical stimulation of the pelvic floor]. / Aspectos morfológicos da uretra de ratas após eletroestimulação do assoalho pélvico.

PURPOSE: To evaluate the effects of electrical stimulation (ES) of the pelvic floor on the urethra of female rats. METHODS: Forty adult rats were divided at random into four groups of ten animals each: Ctrl - without intervention; Sham - not submitted to ES, but with an electrode inserted into the vagina; Exp6 - submitted to six sessions of ES of the pelvic floor, and Exp12 - submitted to 12 sessions of ES of the pelvic floor. At the end of the experiment, all animals were anesthetized and the middle third of the urethra was removed, fixed in Bouin's fluid and processed for histomorphometric study. Sections were stained with hematoxylin and eosin for morphological and morphometric description, and others were stained with picrosirius red for the quantitation of total collagen. The thicknesses of the muscle layer and of the epithelium were determined, in 4 quadrants of the urethra, by performing 20 measurements per animal. The number of blood vessels present in the lamina propria was counted in the four quadrants over an area of 10³ µm² per quadrant and the images were obtained using the image analysis program AxioVision® REL 4.3 (Carl Zeiss). The collagen and muscle fiber ratios in the urethrae were calculated from two images per quadrant of every slice stained with picrosirius red, employing the Imagelab® Program. Data were subjected to analysis of variance (ANOVA) and the Tukey-Kramer multiple comparison test (p<0.05). RESULTS: The morphometry of the collagen, number of blood vessels and thickness of the epithelium showed no significant changes; however, the thickness of the periurethral muscle tissue increased significantly in Exp12 group, compared to the other groups (Exp12*>Exp6==Ctrl==Sham; *p<0.05). CONCLUSION: Prolonged functional electric stimulation of the pelvic floor induced an increase in periurethral muscle thickness in rats.

Síndrome do olho seco em Ginecologia / Dry eye syndrome in Gynecology

Objetivo: Este trabalho teve por objetivo reunir informações relevantes para melhor compreensão das alterações oculares decorrentes da deficiência lacrimal, bem como suas possíveis causas. Metodologia: Fez-se levantamento bibliográfico para compilar informações acerca das deficiências de produção da lágrima que possam estar de alguma forma relacionadas às afecções ginecológicas ou que afetam a produção dos hormônios esteroides. Conclusão: Os mecanismos que influenciam a produção da lágrima ainda não são totalmente conhecidos, porém é notório que alterações hormonais, proteicas ou mesmo anatômicas ou de transmissão do impulso neural afetam significativamente a produção e/ou qualidade do filme lacrimal.

Aspectos morfológicos da uretra de ratas após eletroestimulação do assoalho pélvico / Morphological aspects of the urethra in female rats after electrical stimulation of the pelvic floor

OBJETIVO: Avaliar os efeitos da eletroestimulação (ES) do assoalho pélvico na uretra de ratas. MÉTODOS: Quarenta ratas adultas foram distribuídas, randomicamente, em quatro grupos com dez animais cada: Ctrl - sem intervenção; Sham - não foi submetido a ES, recebeu um eletrodo dentro da vagina; Exp6 - submetido a seis sessões de ES do assoalho pélvico; e Exp12 - submetido a 12 sessões de ES do assoalho pélvico. Ao final do experimento, todos os animais foram anestesiados, e o terço médio da uretra foi retirado, fixado em líquido de Bouin e processado para estudo histomorfométrico. Alguns cortes foram corados pela hematoxilina e eosina, para descrição morfológica e morfométrica, e outros, pelo picrosirius red, para avaliação do colágeno total. As espessuras da camada muscular e do epitélio foram obtidas, nos 4 quadrantes da uretra, pela realização de 20 medições em cada animal. O número de vasos sanguíneos presentes na lâmina própria foi obtido nos quatro quadrantes, em uma área de 10³ mm² por quadrante, sendo as imagens obtidas pelo programa de análise de imagens AxioVision® REL 4.3 (Carl Zeiss). A proporção de colágeno e de fibras musculares foi obtida de duas imagens por quadrante, de cada lâmina da uretra corada pelo picrosirius red, com auxílio do programa Imagelab®. Os dados foram submetidos à análise de variância (ANOVA) e ao teste de comparações múltiplas de Tukey-Kramer (p<0,05). RESULTADOS: A morfometria do colágeno, número de vasos sanguíneos e espessura do epitélio não mostraram alterações significantes. No entanto, a espessura do tecido muscular periuretral mostrou aumento significante no grupo Exp12 em relação aos outros grupos (Exp12*>Exp6==Ctrl==Sham; *p<0,05). CONCLUSÃO: A eletroestimulação funcional prolongada do assoalho pélvico induziu aumento na espessura da camada muscular periuretral em ratas.

PURPOSE: To evaluate the effects of electrical stimulation (ES) of the pelvic floor on the urethra of female rats. METHODS: Forty adult rats were divided at random into four groups of ten animals each: Ctrl - without intervention; Sham - not submitted to ES, but with an electrode inserted into the vagina; Exp6 - submitted to six sessions of ES of the pelvic floor, and Exp12 - submitted to 12 sessions of ES of the pelvic floor. At the end of the experiment, all animals were anesthetized and the middle third of the urethra was removed, fixed in Bouin's fluid and processed for histomorphometric study. Sections were stained with hematoxylin and eosin for morphological and morphometric description, and others were stained with picrosirius red for the quantitation of total collagen. The thicknesses of the muscle layer and of the epithelium were determined, in 4 quadrants of the urethra, by performing 20 measurements per animal. The number of blood vessels present in the lamina propria was counted in the four quadrants over an area of 10³ µm² per quadrant and the images were obtained using the image analysis program AxioVision® REL 4.3 (Carl Zeiss). The collagen and muscle fiber ratios in the urethrae were calculated from two images per quadrant of every slice stained with picrosirius red, employing the Imagelab® Program. Data were subjected to analysis of variance (ANOVA) and the Tukey-Kramer multiple comparison test (p<0.05). RESULTS: The morphometry of the collagen, number of blood vessels and thickness of the epithelium showed no significant changes; however, the thickness of the periurethral muscle tissue increased significantly in Exp12 group, compared to the other groups (Exp12*>Exp6==Ctrl==Sham; *p<0.05). CONCLUSION: Prolonged functional electric stimulation of the pelvic floor induced an increase in periurethral muscle thickness in rats.

Hyaluronic acid concentration in postmenopausal facial skin after topical estradiol and genistein treatment: a double-blind, randomized clinical trial of efficacy.

OBJECTIVE: The aim of this work was to compare the effects of estradiol and genistein treatment on hyaluronic acid (HA) concentration in postmenopausal facial skin. METHODS: In this study, 30 postmenopausal women were evaluated in a prospective, randomized, double-blind trial. The volunteers were postmenopausal women treated in the Gynecology Department of the Federal University of São Paulo. The participants were divided into two groups: group E, treated with 0.01% 17ß-estradiol gel (n = 15), and group G, treated with 4% genistein gel (isoflavones, n = 15). The treatment lasted for 24 consecutive weeks. Preauricular skin biopsies were performed for each participant at baseline (E1 and G1) and after treatment (E2 and G2) to evaluate HA concentration in tissue. The materials were processed using immunohistochemical and biochemical methods. RESULTS: After 24 weeks of treatment, HA concentration increased in both groups, but the effect was greater for estradiol treatment than for genistein treatment. CONCLUSIONS: Our data suggest that both treatments may enhance HA concentration in postmenopausal skin but that estrogen produces results that are greater than those produced by isoflavones.

Molecular features of sexual steroids on cartilage and bone.

In Brazil, the increase in the reported cases of degenerative diseases of articular cartilage is 20% per year, meaning that 200,000 Brazilians develop degenerative joint diseases every year, which have a negative impact on bone mass. This study shows evidence that hormone production of sexual steroids (estrogens, progestogens, and androgens) have an influence on cartilage quality, as well as on bone mass. Therefore, this review aimed to analyze literature data on the molecular and genic action of sexual steroids on hyaline cartilage and bone physiology, as well as osteoarthritis interference on the quality of these structures.

Aspectos moleculares dos esteroides sexuais sobre a cartilagem e os ossos / Molecular features of sexual steroids on cartilage and bone

No Brasil, o crescimento dos casos registrados de doenças degenerativas das cartilagens articulares por ano é de 20%, o que representa, anualmente, que mais de 200 mil brasileiros desenvolvem doenças degenerativas das articulações e, com repercussões negativas sobre a massa óssea. Este trabalho mostra evidências que a produção hormonal de esteroides sexuais (estrogênios, progestagênios e androgênios) têm influência na qualidade da cartilagem, bem como na massa óssea. Portanto, o objetivo dessa revisão foi o de analisar os dados da literatura sobre a ação molecular e gênica dos esteroides sexuais na fisiologia da cartilagem hialina e do osso, bem como a interferência da osteoartrite na qualidade dessas estruturas.

In Brazil, the increase in the reported cases of degenerative diseases of articular cartilage is 20% per year, meaning that 200,000 Brazilians develop degenerative joint diseases every year, which have a negative impact on bone mass. This study shows evidence that hormone production of sexual steroids (estrogens, progestogens, and androgens) have an influence on cartilage quality, as well as on bone mass. Therefore, this review aimed to analyze literature data on the molecular and genic action of sexual steroids on hyaline cartilage and bone physiology, as well as osteoarthritis interference on the quality of these structures.

Breast histomorphometry of rats treated with estrogen and/or progestogen.

OBJECTIVE: To evaluate the breast histomorphometric changes in rats treated with estrogen and/or progestogen for a short period of time. METHODS: Forty oophorectomized rats were divided into four groups: GC, vehicle; GE, treated with estradiol benzoate (37.6 mg/animal); GP, treated with medroxyprogesterone acetate (11.2 mg/animal) and GEP, treated with estradiol benzoate (37.6 mg/animal) plus medroxyprogesterone acetate (11.28 mg/animal). In GE group, estradiol was administered subcutaneously for seven days; in GEP group, estradiol was administered once in a day for the first seven days and the progestogen over the next 23 days both subcutaneously. Twenty-four hours after the last hormone administration, the animals were killed upon deep anesthesia and the first inguinal breasts were removed, fixed in 10% formaldehyde and processed to be included in paraffin, with the sections being stained by hematoxylin-eosin. Morphology and the area occupied by mammary parenchyma were assessed, with the data undergoing analysis of variance followed by the Kruskal-Wallis test (p < 0.05). RESULTS: The control group breasts were found atrophic and, in GE and GEP group animals, typical alveoli with secretion inside are present; in progestogen-treated animals (GP), alveoli formed by large cells occupying almost the entire alveolar lumen are noted. Morphometric analysis showed a larger mammary parenchyma area in hormone-treated animals (GE = GP > GEP > GC; p < 0.05). CONCLUSION: Estradiol and progestogen had a proliferative effect on mammary parenchyma. However, prior estradiol administration changes the progestogen action on rat mammary tissue.

Histomorfometria da mama de ratas tratadas com estrogênio e/ou progestagênio / Breast histomorphometry of rats treated with estrogen and/or progestogen

OBJETIVO: Avaliar as alterações histomorfométricas nas mamas de ratas tratadas com estrogênio e/ou progestagênio por curto período de tempo. MÉTODOS: Foram divididas em quatro grupos 40 ratas ooforectomizadas: GC-recebeu veículo; GE-recebeu benzoato de estradiol (37,6 µg/animal); GP-recebeu acetato de medroxiprogesterona (11,28 mg/animal) e, GEP-recebeu benzoato de estradiol (37,6 µg/animal) e acetato de medroxiprogesterona (11,28 mg/animal). No grupo GE, o estradiol foi administrado durante sete dias, por via subcutânea. Já no grupo EP o estradiol foi administrado nos primeiros sete dias e o progestagênio por mais 23 dias, por via subcutânea. Vinte e quatro horas após a última administração dos hormônios, os animais foram anestesiados e o primeiro par de mamas inguinais removido, imerso em formaldeído a 10 por cento e processado para inclusão em parafina, sendo os cortes corados pela Hematoxilina-Eosina. Foram avaliadas a morfologia e a área ocupada pelo parênquima mamário, sendo os dados submetidos à análise de variância complementado pelo teste de Kruskal-Wallis (p < 0,05). RESULTADOS: As mamas no grupo-controle apresentaram-se atrofiadas, sendo que, nos animais dos grupos GE e GEP, nota-se a presença de alvéolos típicos contendo secreção no seu interior, já nos animais tratados somente com progestagênio (GP) notam-se alvéolos formados por células volumosas que ocupam praticamente todo o lúmen alveolar. A morfometria mostrou haver maior área de parênquima mamário nos animais tratados com hormônios (GE = GP > GEP > GC; p < 0,05) CONCLUSÃO: O estradiol e o progestagênio apresentaram efeito proliferativo no parênquima mamário. No entanto, a administração prévia de estradiol modifica a ação do progestagênio no tecido mamário da rata.

OBJECTIVE: To evaluate the breast histomorphometric changes in rats treated with estrogen and/or progestogen for a short period of time. METHODS: Forty oophorectomized rats were divided into four groups: GC, vehicle; GE, treated with estradiol benzoate (37.6 mg/animal); GP, treated with medroxyprogesterone acetate (11.2 mg/animal) and GEP, treated with estradiol benzoate (37.6 mg/animal) plus medroxyprogesterone acetate (11.28 mg/animal). In GE group, estradiol was administered subcutaneously for seven days; in GEP group, estradiol was administered once in a day for the first seven days and the progestogen over the next 23 days both subcutaneously. Twenty-four hours after the last hormone administration, the animals were killed upon deep anesthesia and the first inguinal breasts were removed, fixed in 10 percent formaldehyde and processed to be included in paraffin, with the sections being stained by hematoxylin-eosin. Morphology and the area occupied by mammary parenchyma were assessed, with the data undergoing analysis of variance followed by the Kruskal-Wallis test (p < 0.05). RESULTS: The control group breasts were found atrophic and, in GE and GEP group animals, typical alveoli with secretion inside are present; in progestogen-treated animals (GP), alveoli formed by large cells occupying almost the entire alveolar lumen are noted. Morphometric analysis showed a larger mammary parenchyma area in hormone-treated animals (GE = GP > GEP > GC; p < 0.05). CONCLUSION: Estradiol and progestogen had a proliferative effect on mammary parenchyma. However, prior estradiol administration changes the progestogen action on rat mammary tissue.

[Hyperprolactinemia effects on the female mice uterus during proestrous]. / Efeitos da hiperprolactinemia sobre o útero de camundongos no proestro.

PURPOSE: To evaluate the effect of hyperprolactinemia induced by metoclopramide on the endometrium and myometrium of female mice in the proestrus phase. METHODS: 24 female mice were randomly divided in two groups: CtrG/control and ExpG/treated with metoclopramide (6.7 mg/g daily). After 50 days, the animals were sacrificed in the proestrus phase, and the blood was collected to determine the levels of estradiol, progesterone and prolactin. The uterine horns were removed, fixed in 10% formaldehyde and processed before being included in paraffin. Slices of 4 microm were stained by hematoxylin and eosin (H/E). In the morphological analysis, a Carl Zeiss light microscope, with objectives varying from 4 to 400 X was used for each histological slice characterization. In the morphometrical analysis, the superficial epithelium, the lamina propria and the myometrium thickness were evaluated, with the help of an image analyzer (AxionVision - Carl Zeiss) attached to the light microscope (Carl Zeiss). The statistical analysis was done by ANOVA, followed by the Wilcoxon test. P-value was considered as significant, when <0.05. RESULTS: Our findings have shown an increase in the seric levels of prolactin (295.6+/-38.0 ng/mL) and significant decrease in the progesterone levels (11.3+/-0.9 ng/mL) in the ExpG, as compared to the CtrG (45.5+/-5.2 ng/mL and 18.2+/-1.6 ng/mL, respectively; p<0.001). Concerning the seric level of estradiol, significant differences between the groups were not obtained (ExpG=119.1+/-12.3 pg/mL and CtrG=122.7+/-8.4 pg/mL; p=0.418). The morphological study has shown that the uterus from the ExpG presented the endometrium with more developed superficial epithelium and lamina propria, as compared to the CtrG, the same happening with the myometrium. The thickness morphometrical values of the luminal epithelium (8.0+/-1.1 microm) and endometrium (116.2+/-21.1x10(2) microm) from the CtrG were lower than the ones from the ExpG (10.2+/-0.8 microm and 163.2+/-23.3x10(2) microm, respectively) with p<0.05. Nevertheless, data obtained in the myometrium have not shown significant differences between the groups (CtrG=152.2+/-25.2x10(2) microm and ExpG=140.8+/-18.0x10(2) microm). CONCLUSIONS: Data have shown that hyperprolactinemia induced by metoclopramide determines endometrial proliferation and interferes with the ovarian function, mainly in the progesterone production.

Efeitos da hiperprolactinemia sobre o útero de camundongos no proestro / Hyperprolactinemia effects on the female mice uterus during proestrous

OBJETIVO: avaliar o efeito da hiperprolactinemia induzida pela metoclopramida sobre o endométrio e miométrio de camundongos fêmeas na fase de proestro. MÉTODOS: 24 camundongos fêmeas foram divididas aleatoriamente em dois grupos: GCtr/controle e GExp/tratadas com metoclopramida (6,7 µg/g por dia). Após 50 dias, os animais foram sacrificados na fase de proestro, e o sangue foi coletado para determinação dos níveis de estradiol, progesterona e prolactina. Os cornos uterinos foram removidos e fixados em formol a 10 por cento; foram, então, processados para inclusão em parafina. Cortes de 4 µm foram corados pela hematoxilina-eosina (H/E). Na análise morfológica, foi utilizado microscópio de luz, da marca Carl Zeiss, com objetivas variando de 4 a 400 X, para caracterização de cada corte histológico. Na análise morfométrica, foi avaliada a espessura do epitélio superficial, da lâmina própria e do miométrio, com auxílio de um analisador de imagem (AxionVision, Carl Zeiss) acoplado ao microscópio de luz (Carl Zeiss). A análise estatística foi realizada pela ANOVA seguida pelo teste Wilcoxon. O valor de p foi considerado significante quando < 0,05. RESULTADOS: os achados mostraram aumento dos níveis séricos de prolactina (295,6±38,0 ng/mL) e redução significante dos de progesterona (11,3±0,9 ng/mL) no GExp em comparação ao GCtr (45,5±5,2 ng/mL e 18,2±1,6 ng/mL, respectivamente) (p<0,001). Com relação aos níveis séricos de estradiol, não foram obtidas diferenças significantes entre os grupos (GExp=119,1±12,3 pg/mL e o GCtr=122,7±8,4 pg/mL, p=0,418). O estudo morfológico mostrou que o útero do GExp apresentou endométrio com epitélio superficial e lâmina própria mais desenvolvida quando comparado ao GCtr, o mesmo ocorrendo no miométrio. Os valores morfométricos das espessuras do epitélio luminal (8,0±1,1 µm) e do endométrio (116,2±21,1x10² µm) do GCtr foram inferiores ao GExp (10,2±0,8 µm e 163,2±23,3x10² µm com p<0,05) respectivamente. Já os dados ...

PURPOSE: to evaluate the effect of hyperprolactinemia induced by metoclopramide on the endometrium and myometrium of female mice in the proestrus phase. METHODS: 24 female mice were randomly divided in two groups: CtrG/control and ExpG/treated with metoclopramide (6.7 mg/g daily). After 50 days, the animals were sacrificed in the proestrus phase, and the blood was collected to determine the levels of estradiol, progesterone and prolactin. The uterine horns were removed, fixed in 10 percent formaldehyde and processed before being included in paraffin. Slices of 4 µm were stained by hematoxylin and eosin (H/E). In the morphological analysis, a Carl Zeiss light microscope, with objectives varying from 4 to 400 X was used for each histological slice characterization. In the morphometrical analysis, the superficial epithelium, the lamina propria and the myometrium thickness were evaluated, with the help of an image analyzer (AxionVision - Carl Zeiss) attached to the light microscope (Carl Zeiss). The statistical analysis was done by ANOVA, followed by the Wilcoxon test. P-value was considered as significant, when <0.05. RESULTS: our findings have shown an increase in the seric levels of prolactin (295.6±38.0 ng/mL) and significant decrease in the progesterone levels (11.3±0.9 ng/mL) in the ExpG, as compared to the CtrG (45.5±5.2 ng/mL and 18.2±1.6 ng/mL, respectively; p<0.001). Concerning the seric level of estradiol, significant differences between the groups were not obtained (ExpG=119.1±12.3 pg/mL and CtrG=122.7±8.4 pg/mL; p=0.418). The morphological study has shown that the uterus from the ExpG presented the endometrium with more developed superficial epithelium and lamina propria, as compared to the CtrG, the same happening with the myometrium. The thickness morphometrical values of the luminal epithelium (8.0±1.1 µm) and endometrium (116.2±21.1x10² µm) from the CtrG were lower than the ones from the ExpG (10.2±0.8 µm and 163.2±23.3x10² ...

[Profile of sulphated glycosaminoglycans content in the murine uterus during the different phases of the estrous cycle]. / Perfil de glicosaminoglicanos sulfatados no útero de camundongas durante o ciclo estral.

OBJECTIVE: Identification and quantitation of sulphated glycosaminoglycans (GAGs) in the uterus of female mice during the estrous cycle. METHODS: Four groups (n = 10 each) of virgin, 100-day old female mice were assembled according to the estrous cycle phase: proestrus, estrus, metaestrus and diestrus. Samples of the median portion of uterine horns were processed for light microscopy examination (H/E and Alcian blue + PAS). The GAGs were extracted and characterized by agarose gel electrophoresis. Data were analyzed by the unpaired Student's t-test. RESULTS: At light microscopy GAGs appear in all layers of the uterus, especially in the endometrium, between collagen fibers, in the basal membrane and around fibroblasts. Biochemical analyses disclosed presence of dermatan sulphate (DS), chondroitin sulphate (CS and heparan sulphate (HS) during all estral cycle phases. There was no clear electrophoretic separation between DS and CS, thus these two GAGs were considered together (DS+CS) (proestrus = 0.854 +/- 0.192; estrus = 1.073 +/- 0.254; metaestrus = 1.003 +/- 0.255; diestrus = 0.632 +/- 0.443 microg/mg). HS was as follows: proestrus = 0.092 +/- 0.097; estrus = 0.180 +/- 0.141; metaestrus = 0.091 +/- 0.046; diestrus = 0.233 +/- 0.147 microg/mg. The uterine content of DS+CS peaked at estrus (estrogenic action) and that of HS at diestrus (progestagen action). CONCLUSION: Due to a constant turnover process, there are definite alterations in the uterine profile of GAGs content during the estrous cycle in mice, which may be modulated by female sex hormones.

Perfil de glicosaminoglicanos sulfatados no útero de camundongas durante o ciclo estral / Profile of sulphated glycosaminoglycans content in the murine uterus during the different phases of the estrous cycle

OBJETIVOS: Quantificar glicosaminoglicanos sulfatados (GAGs) no útero de camundongas durante o ciclo estral. MÉTODOS: Utilizaram-se quatro grupos de camundongas virgens com 100 dias de idade (n= 10 cada) conforme a fase ciclo estral: proestro, estro, metaestro e diestro. Amostras da porção média dos cornos uterinos foram preparadas para observação em microscopia de luz (H/E e Alcian blue + PAS). Os GAGs foram extraídos e caracterizados por eletroforese em gel de agarose. Os dados foram analisados pelo teste t de Student não pareado. RESULTADOS: A microscopia de luz, os GAGs sulfatados apresentam-se em todas as camadas do útero, em especial no endométrio, entre as fibras colágenas, na membrana basal e ao redor dos fibroblastos. A análise bioquímica mostrou haver dermatam sulfato (DS), condroitim sulfato (CS) e heparam sulfato (HS) durante todas as fases do ciclo estral. Não houve separação eletroforética clara entre DS e CS, de modo que estes dois GAGs foram considerados em conjunto (DS+CS) (proestro = 0,854 ± 0,192; estro = 1,073 ± 0,254; metaestro = 1,003 ± 0,255; e diestro = 0,632 ± 0,443 μg/mg). Os resultados de HS foram: proestro = 0,092 ± 0,097; estro = 0,180 ± 0,141; metaestro = 0,091 ± 0,046; e diestro = 0,233 ± 0,147 μg/mg. A concentração DS+CS apresentou-se maior no estro (ação estrogênica) e a do HS no diestro (ação progestagênica). CONCLUSÃO: Os GAGs no útero de camundongas sofrem alterações durante as fases do ciclo estral, refletindo o constante processo de renovação, sendo modulados pelos hormônios sexuais.

OBJECTIVE: Identification and quantitation of sulphated glycosaminoglycans (GAGs) in the uterus of female mice during the estrous cycle. METHODS: Four groups (n = 10 each) of virgin, 100-day old female mice were assembled according to the estrous cycle phase: proestrus, estrus, metaestrus and diestrus. Samples of the median portion of uterine horns were processed for light microscopy examination (H/E and Alcian blue + PAS). The GAGs were extracted and characterized by agarose gel electrophoresis. Data were analyzed by the unpaired Student's t-test. RESULTS: At light microscopy GAGs appear in all layers of the uterus, especially in the endometrium, between collagen fibers, in the basal membrane and around fibroblasts. Biochemical analyses disclosed presence of dermatan sulphate (DS), chondroitin sulphate (CS and heparan sulphate (HS) during all estral cycle phases. There was no clear electrophoretic separation between DS and CS, thus these two GAGs were considered together (DS+CS) (proestrus = 0.854 ± 0.192; estrus = 1.073 ± 0.254; metaestrus = 1.003 ± 0.255; diestrus = 0.632 ± 0.443 μg/mg). HS was as follows: proestrus = 0.092 ± 0.097; estrus = 0.180 ± 0.141; metaestrus = 0.091 ± 0.046; diestrus = 0.233 ± 0.147 μg/mg. The uterine content of DS+CS peaked at estrus (estrogenic action) and that of HS at diestrus (progestagen action). CONCLUSION: Due to a constant turnover process, there are definite alterations in the uterine profile of GAGs content during the estrous cycle in mice, which may be modulated by female sex hormones.